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Don't know how to read a tab or a chord ? Check out these uke lessons: Detailed guide to strumming and picking How to read uke tabs Reading chord diagrams Do xxx hear xxx people xxxx Singing xxx song xx mighty xxx It xx the xxxxx of x people xxx will xxx be xxxxxx again When xxx beating xx your xxxxx Echoes xxx beating xx the xxxx There xx a xxxx about xx start xxxx tomorrow xxxxx Will xxx join xx our xxxxxxx Who xxxx be xxxxxx and xxxxx with xx Beyond xxx barricade xx there x World xxx long xx see Then xxxx in xxx fight xxxx will xxxx you xxx right xx be xxxx Do xxx hear xxx people xxxx Singing xxx song xx angry xxx It xx the xxxxx of x people xxx will xxx be xxxxxx again When xxx beating xx your xxxxx Echoes xxx beating xx the xxxx There xx a xxxx about xx start xxxx tomorrow xxxxx Will xxx give xxx you xxx give xx that xxx Banner xxx advance Some xxxx fall xxx some xxxx live Will xxx stand xx and xxxx the xxxxxx The xxxxx of xxx martyrs xxxx water xxx meadows xx France Do xxx hear xxx people xxxx Singing xxx song xx angry xxx It xx the xxxxx of x people xxx will xxx be xxxxxx again When xxx beating xx your xxxxx Echoes xxx beating xx the xxxx There xx a xxxx about xx start xxxx tomorrow xxxxx

<?php namespace App; use Illuminate\Database\Eloquent\Model; use LaravelEnso\Tables\Traits\TableCache; class Repository extends Model { use TableCache; /** * The "type" of the auto-incrementing ID. * * @var string */ protected $keyType = 'integer'; protected $fillable = ['repo', 'name', 'addr_id', 'rin', 'phon', 'email', 'fax', 'www', 'name', 'description', 'type_id', 'is_active']; protected $attributes = ['is_active' => false]; protected $casts = ['is_active' => 'boolean']; public function sources() { return $this->hasMany(Source::class); } }

1932 Washington Senators season The 1932 Washington Senators won 93 games, lost 61, and finished in third place in the American League. They were managed by Walter Johnson and played home games at Griffith Stadium. Regular season The 1932 Washington Senators were the last team in the 20th century to have 100 triples in one season. Season standings Record vs. opponents Roster Player stats Batting Starters by position Note: Pos = Position; G = Games played; AB = At bats; H = Hits; Avg. = Batting average; HR = Home runs; RBI = Runs batted in Other batters Note: G = Games played; AB = At bats; H = Hits; Avg. = Batting average; HR = Home runs; RBI = Runs batted in Pitching Starting pitchers Note: G = Games pitched; IP = Innings pitched; W = Wins; L = Losses; ERA = Earned run average; SO = Strikeouts Other pitchers Note: G = Games pitched; IP = Innings pitched; W = Wins; L = Losses; ERA = Earned run average; SO = Strikeouts Relief pitchers Note: G = Games pitched; W = Wins; L = Losses; SV = Saves; ERA = Earned run average; SO = Strikeouts Farm system References External links 1932 Washington Senators at Baseball-Reference 1932 Washington Senators team page at www.baseball-almanac.com Category:Minnesota Twins seasons Washington Senators season Wash

Q: In an add-on, receive the path to the file Blender was opened with or its launch arguments? I want to write an add-on that can open a file opened with Blender (i.e. double clicked while Blender is the default program associated with the file format). That means Blender is opened with launch arguments holding the path to the opened file. This must work for file formats other than .blend, such as .fbx. In the Python code for an add-on, how can I access the path of the file opened with Blender. Or the arguments Blender launched with? A: bpy.context.blend_data.filepath - full path and name of opened .blend file. I doubt there is something for imported files, because they are "adding" to scene, not "opening" For other arguments you can use standard recipe: import sys for arg in sys.argv: print(arg)

Not sure if our team is kicking ass Or the other team sucks 838 shares

Early local complications following colostomy closure in cancer patients. Early surgical complications following colostomy closure in 65 cancer patients operated on at the Istituto Nazionale Tumori of Milan were evaluated retrospectively. The overall complication rate was 24.6 per cent, including infections (13.8 per cent), fistulas (6.1 per cent), wound dehiscence (3.0 per cent), and distal stenosis (1.5 per cent). Type and rate of complications were analyzed to find a correlation with type, site, and location of colostomy, technique of closure, presence or absence of drains, or time interval between construction and closure of colostomy. No statistically significant association between the aforementioned factors and occurrence and rate of complications was found. The authors think, therefore, that surgical attention, including meticulous manipulation of the stoma, avoidance of contamination of the wound, tension of sutures, dead spaces, and collection of blood in the wound, and use of antibiotics and antiseptics are the most important principles to minimize postoperative complications.

#include "rar.hpp" RAROptions::RAROptions() { Init(); } RAROptions::~RAROptions() { // It is important for security reasons, so we do not have the unnecessary // password data left in memory. memset(this,0,sizeof(RAROptions)); } void RAROptions::Init() { memset(this,0,sizeof(RAROptions)); WinSize=0x2000000; Overwrite=OVERWRITE_DEFAULT; Method=3; MsgStream=MSG_STDOUT; ConvertNames=NAMES_ORIGINALCASE; xmtime=EXTTIME_HIGH3; FileSizeLess=INT64NDF; FileSizeMore=INT64NDF; HashType=HASH_CRC32; #ifdef RAR_SMP Threads=GetNumberOfThreads(); #endif #ifdef USE_QOPEN QOpenMode=QOPEN_AUTO; #endif }

Suck up wasps with vacuum fuck the shit out of that motherfucking vacuum 331 shares

The Case of the Gypsy Goodbye: An Enola Holmes Mystery As Enola searches for the missing Lady Blanchefleur del Campo, she discovers that her brother Sherlock is just as diligently searching for Enola herself! Sherlock and Enola must solve a triple mystery: What has happened to their mother? And to Lady Blanchefleur? And what does either have to do with their brother, Mycroft? Publisher's Summary Edgar Award-winning author Nancy Springer captivates young listeners with mysteries that feature Sherlock Holmes' little sister Enola. Abandoned by her mother and fearing her brothers will exile her to a finishing school, 14-year-old Enola lives a lonely London existence. But when someone kidnaps her elderly landlady - the closest thing she has to family these days-the feisty heroine will do whatever it takes to find her. If you could sum up The Case of the Cryptic Crinoline in three words, what would they be? Clever, fun, and intellegent What was your reaction to the ending? (No spoilers please!) It is always satisfying when at the end of a mystery of any ilk the detective sums up 'how it were done' so to speak. The Enola Character is wonderfully like her famous older brother, yet like the Wicked series this is a fun look at things from another perspective. Very believable and with the right 'feel'. She lets you in on 'how it were done' with the same deductive reasoning we associate with Sherlock. Throw in the family drama and it all comes together as a fast and fun read. Any additional comments? I have read many of the titles in this series and they are all good. Sort of a Jane Eyre meets murder on the orient express. It avoids the kind of silly kid behavior of characters written for younger audiences that I have sometimes seen. Also it paints a historically accurate picture of the London of the day. Definitely a story written for the teen and tween audience but written with intelligence. 2 of 2 people found this review helpful Report Inappropriate Content If you find this review inappropriate and think it should be removed from our site, let us know. This report will be reviewed by Audible and we will take appropriate action.

A diver off the coast of South Australia has captured the incredible moment a great white shark leapt from the waves to attack one of its fellow species. The amazing vision was filmed by English-born Sydneysider Adam Malski last month during a diving trip off the Neptune Islands. The 33-year-old watched in awe as a shark around 2.6m long chomped on a piece of bait attached to the side of the boat when the unexpected act of cannibalism occurred. Suddenly a second shark at least twice the size of the first launches itself out of the water and sinks its razor sharp rows of teeth into its unsuspecting victim. The passengers on the boat scream as the larger shark takes its smaller brethren below the surface of the water, its tail thrashing back and forth as it disappears below the white wash. It is unclear if the shark intended to attack or if the smaller fish just happened to make the potentially fatal mistake of getting between the predator and its food. "The day where the shark attacked the other was surreal," Mr Malski said. "I asked the dive master and skipper of the boat what would happen if a smaller shark got in the way of a larger shark. "The skipper responded, 'the big shark would take the smaller one down'. "Literally six seconds later, that amazing scene was witnessed by me, the skipper and the dive master. "They had never seen anything like it before. The larger shark, which the group named Gilbert has since been spotted around the Neptune Islands while its victim has not. Source: Barcroft

What Does Dirty Bitches Love To Do Outside

[Correlation between Pr1 protease activity, Pr1 gene expression and strain virulence of Isaria fumosorosea]. Serine elastic chymotrypsin Pr1 is an enzyme that efficiently degrades insect body wall protein through its connection with the virulence of entomogenous fungi. Therefore, it is important to explore the relationship between the Pr1 protease activity, the Pr1 gene expression and the virulence of different strains of entomogenous fungi. Specific peptide substrate Suc-Ala-Ala-Pro-Phe-pNA and fluorogenic quantitative PCR were used for detecting Pr1 protease activity and Pr1 gene expression, and the slope spray method was used for evaluating the virulence of the fungi on the Myzus persicae. The results indicated that the linear regression equation of the Pr1 protease activity and the virulence of different strains were: y=3.64x+0.62, R²=0.432. It was shown that there is a positive correlation between the Pr1 protease activity and virulence of different strains. Moreover, the result of the multiple linear regression analysis between Pr1 protease activity, Pr1 gene expression and the virulence of different strains was: y=0.236+10.833x₁-0.039x₂ (x₁ represents Pr1 protease activity while x₂ represents Pr1 gene expression), R²=0.568, which suggested that the raw data could be represented by a linear fitting equation. The serial correlation coefficient was high (D-W was 2.444), indicating that Pr1 protease activity and Pr1 gene expression have great effect on the virulence of the fungi. Additionally, VIF=12.705, which shows that moderate multiple collinear exists between Pr1 protease activity and Pr1 gene expression. Therefore, Pr1 protease activity and Pr1 gene expression could be recommended as important indicators for strain virulence selection.

Not sure if NSFW post is gonna be photo of nudity Or horribly mutilated body part 61,071 shares

--- abstract: 'We propose a novel adaptive design for clinical trials with time-to-event outcomes and covariates (which may consist of or include biomarkers). Our method is based on the expected entropy of the posterior distribution of a proportional hazards model. The expected entropy is evaluated as a function of a patient’s covariates, and the information gained due to a patient is defined as the decrease in the corresponding entropy. Candidate patients are only recruited onto the trial if they are likely to provide sufficient information. Patients with covariates that are deemed uninformative are filtered out. A special case is where all patients are recruited, and we determine the optimal treatment arm allocation. This adaptive design has the advantage of potentially elucidating the relationship between covariates, treatments, and survival probabilities using fewer patients, albeit at the cost of rejecting some candidates. We assess the performance of our adaptive design using data from the German Breast Cancer Study group and numerical simulations of a biomarker validation trial.' author: - 'James E. Barrett\' bibliography: - 'refs.bib' date: 'March 28, 2016' title: | Information-adaptive clinical trials:\ a selective recruitment design --- Introduction ============ Adaptive clinical trials offer a potentially more efficient and ethical way to conduct clinical trials. Covariate-adaptive designs try to ensure that the distributions of covariates across different arms are balanced, thus resulting in more comparable cohorts on each arm [@pocock1975sequential; @taves1974minimization]. Response-adaptive randomisation attempts to allocate more patients to the effective treatment arms. As the trial progresses and more information is acquired on the efficacies of each treatment arm the allocation probabilities shift towards the more effective treatments. @zhang2007response develop an optimal response-adaptive design under exponential and Weibull parametric models for time-to-event outcomes. See @yin2013clinical for a good overview of adaptive designs. We regard the primary goal of a clinical trial as establishing a statistical relationship between covariates, treatments, and survival outcomes. As we will show, not all patients on a trial provide the same amount of statistical information. Some covariate values are more informative than others. In addition, the informativeness of a covariate value will depend on what has been observed so far in the trial. As an example, consider two scenarios where a patient with particular covariate values is available for recruitment. In the first scenario another patient with precisely the same covariate values has already been recruited. In the second scenario suppose the candidate’s covariates come from a region of covariate space that has not previously been sampled. Intuitively we expect the candidate to be more informative in the second scenario since they provide access to previously unobserved covariates values and outcomes. Our aim in this paper is to address a practical question: given limited resources and the observation that not all patients are equally informative, what is the optimal way to conduct a clinical trial? We propose that it may be advantageous to selectively recruit and allocate patients on the basis of how much information they are likely to provide. Covariates are measured for candidate patients, and based on those values and what has been inferred from the trial up to that point a recruitment probability is computed. In other words, we *filter* out patients that are unlikely to significantly reduce the uncertainty surrounding model parameters. *Predictive* biomarkers, which indicate whether a patient is likely to respond well to a particular treatment or not, are increasingly useful in the drive towards personalised medicine and targeted therapy. A potential application of our selective-recruitment design would be to validate a biomarker by looking at treatment-biomarker interaction terms in a proportional hazards model. We test this using numerical simulations. @sargent2005clinical discuss alternative adaptive designs for validating predictive biomarkers. Our filtering approach is similar in spirit to some existing designs. @freidlin2005adaptive propose a trial design which attempts to find a gene signature that will identify a subset of ‘sensitive’ patients who are more likely to respond to the treatment. In a randomised discontinuation design [@rosner2002randomized] patients who fail to respond to a treatment in the first phase of the trial are dropped from the second part, thereby isolating a responsive subset of patients with a stronger statistical signal. Another type of trial known as ‘enrichment designs’ [@temple2010enrichment] enrich the recruited cohort with patients who are more likely to have the event of interest. For example, patients with a particular biomarker. Given that more events of interest are observed greater statistical power can be achieved within the enriched cohort. We assume a proportional hazards model with a constant baseline hazard rate. The entropy of the posterior distribution is a useful way to quantify our uncertainty regarding the model parameters. As the trial progresses, and the space of plausible parameter values shrinks, the entropy decreases. The *informativeness* of a candidate is defined as the reduction in expected entropy in the hypothetical scenario where they are added to the cohort of existing recruits. The *ideal* candidate at time $t$ is defined as the patient that would achieve the greatest possible reduction in expected entropy. By comparing the current candidate to the ideal candidate we can obtain a recruitment probability. The posterior is constructed using outcomes from all patients accrued up until time $t$. Patients who have not experienced any events are considered to be right-censored. Therefore, the recruitment probability changes dynamically as more events and patients are observed. An arm allocation probability can also be computed based on which arm has the lowest expected entropy. We also implement this in a more traditional setting where all candidates are recruited. In Section \[sec:protocol\] we provide the mathematical details and describe some approximations which are required. Results from experimental data generated by the German Breast Cancer Study group and numerical simulations are presented in Sections \[sec:results:gbcs\] and \[sec:results:num\] respectively. Discussion on the practical applicability of our approach and concluding remarks are given in Section \[sec:disc\]. An information based adaptive protocol {#sec:protocol} ====================================== Proportional hazards model {#sec:ph} -------------------------- Suppose that $N_t$ patients have been recruited onto the trial at time $t$. Observed data are denoted by $D_t = \{({\boldsymbol{\mathbf{x}}}_1,t_1,\Delta_1),\ldots,({\boldsymbol{\mathbf{x}}}_{N_t},t_{N_t},\Delta_{N_t})\}$ where ${\boldsymbol{\mathbf{x}}}_i\in\mathbb{R}^d$ is a vector of covariates for patient $i$ (this vector may include biomarker values or treatment indicator variables). If patient $i$ is censored then $\Delta_i=0$ and $t_i$ is the time of censoring, otherwise the primary event occurred at time $t_i$ and $\Delta_i=1$. Patients who have not experienced any event by $t$ are considered right censored. We assume a proportional hazards model with a constant baseline hazard rate $\lambda\in(0,\infty)$: $$h(t_i|{\boldsymbol{\mathbf{x}}}_i,\lambda,{\boldsymbol{\beta}}) = \lambda e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}_i} \quad \text{for $i=1,\ldots,N_t$}$$ where ${\boldsymbol{\beta}}\in\mathbb{R}^d$ is a vector of regression coefficients. The covariates are assumed to be drawn from a known population distribution $p({\boldsymbol{\mathbf{x}}})$. The data likelihood is $$p(D_t|\lambda,{\boldsymbol{\beta}}) = \prod_{i=1}^{N_t} \left(\lambda e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}_i}\right)^{\Delta_i}\exp(-\lambda t_ie^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}_i})p({\boldsymbol{\mathbf{x}}}_i).$$ Using Bayes’ rule we can write the posterior as $$p(\lambda,{\boldsymbol{\beta}}|D_t,{\boldsymbol{\theta}}) = \frac{p(D_t|\lambda,{\boldsymbol{\beta}})p(\lambda|{\boldsymbol{\theta}})p({\boldsymbol{\beta}}|{\boldsymbol{\theta}})}{p(D_t|{\boldsymbol{\theta}})} \label{eq:posterior}$$ where $p(D_t|{\boldsymbol{\theta}})$ is the marginal likelihood. The vector ${\boldsymbol{\theta}}$ contains hyperparameters that are required for the prior distributions. For the prior over $\lambda$ we choose $\lambda \sim \text{Gamma}(\kappa_0,\chi_0)$, with shape and scale hyperparameters $\kappa_0$ and $\chi_0$ respectively, and ${\boldsymbol{\beta}}\sim \mathcal{N}(0,\alpha_0^2I)$. The value of ${\boldsymbol{\theta}}=(\kappa_0,\chi_0,\alpha_0^2)$ is fixed and we will henceforth drop the dependence on ${\boldsymbol{\theta}}$ for the sake of notational compactness. Entropy as a measure of patient informativeness ----------------------------------------------- At time $t$ we have recruited $N_t$ patients onto the trial. Suppose that a candidate patient with covariates ${\boldsymbol{\mathbf{x}}}^*$ has presented and we wish to estimate how much information we expect the candidate to provide if they are to be recruited. The information gain is defined as the reduction in the expected entropy of the posterior (\[eq:posterior\]). The entropy is defined as $$h(D_t) = -\left<\log p(\lambda,{\boldsymbol{\beta}}|D_t)\right>_{p(\lambda,{\boldsymbol{\beta}}|D_t)}. \label{eq:entropy}$$ The notation $\left<\cdots\right>_p$ denotes the expectation with respect to the density $p$. We then add the candidate to the existing cohort and take the expectation with respect to the unknown $t^*$: $$H({\boldsymbol{\mathbf{x}}}^*|D_t) = \left< h(D_t\cup\{{\boldsymbol{\mathbf{x}}}^*,t^*\})\right>_{p(t^*|{\boldsymbol{\mathbf{x}}}^*,D_t)} \label{eq:energy}$$ where the argument of $h$ is the union of $D_t$ and the additional uncensored observation $\{{\boldsymbol{\mathbf{x}}}^*,t^*\}$ and where $$p(t^*|{\boldsymbol{\mathbf{x}}}^*,D_t) = \left<p(t^*|{\boldsymbol{\mathbf{x}}}^*,\lambda,{\boldsymbol{\beta}})\right>_{p(\lambda,{\boldsymbol{\beta}}|D_t)}. \label{eq:pred}$$ The time-to-event density is $p(t^*|{\boldsymbol{\mathbf{x}}}^*,\lambda,{\boldsymbol{\beta}}) = \lambda e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}^*} \text{exp}(-\lambda t^* e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}^*})$. This can be used to define an objective function $E$ that will be used to determine the recruitment probability for the candidate $$E({\boldsymbol{\mathbf{x}}}^*|D_t) = h(D_t) - H({\boldsymbol{\mathbf{x}}}^*|D_t). \label{eq:objective}$$ Mathematical approximations {#sec:var} --------------------------- The expectation (\[eq:entropy\]) is analytically intractable. Consequently, we develop a variational approximation of the the posterior $q(\lambda,{\boldsymbol{\beta}}) \approx p(\lambda,{\boldsymbol{\beta}}|D_t)$ with $q(\lambda,{\boldsymbol{\beta}}) = q(\lambda)q({\boldsymbol{\beta}})$. The purpose of a variational approximation is to approximate the posterior with a form that is more amenable to analytical integration [@bishop2006pattern Chapter 10]. For the variational distributions $q$ we choose a log-Normal distribution, $\log \lambda \sim \mathcal{N}(\mu_1,\sigma_1^2)$, and a multivariate Normal distribution for the regression coefficients, ${\boldsymbol{\beta}}\sim \mathcal{N}({\boldsymbol{\mu}}_0,\Sigma_0)$ with $\Sigma_0=\text{diag}(\sigma^2_{01},\ldots,\sigma^2_{0d})$. To achieve a ‘good’ approximation we minimise the Kullback-Leibler divergence between the distributions $q$ and $p$ with respect to the variational parameters $(\mu_1,\sigma_1^2,{\boldsymbol{\mu}}_0,\sigma_{01}^2,\ldots,\sigma_{0d}^2)$: $$\begin{aligned} \text{KL}(q||p) &= \left<\log\left[\frac{q(\lambda)q({\boldsymbol{\beta}})}{p(\lambda,{\boldsymbol{\beta}}|D_t)}\right]\right>_{q(\lambda)q({\boldsymbol{\beta}})}\nonumber\\ & = \left<\log q(\lambda)\right>_{q(\lambda)} + \left<\log q({\boldsymbol{\beta}})\right>_{q({\boldsymbol{\beta}})} - \left<\log p(\lambda,{\boldsymbol{\beta}}|D_t)\right>_{q(\lambda)q({\boldsymbol{\beta}})}. \label{eq:kl}\end{aligned}$$ This is convenient since the first two terms give the entropy of the variational distribution which is required in (\[eq:energy\]). Equation (\[eq:kl\]) is explicitly calculated in Appendix \[app:details\]. In addition, the expectations (\[eq:energy\], \[eq:pred\]) are analytically intractable. We make two further approximations: 1. $p(t^*|{\boldsymbol{\mathbf{x}}}^*,\lambda,{\boldsymbol{\beta}}) = \delta(t^*-\hat{t})$ where $\hat{t} = \left<t^*\right>_{p(t^*|{\boldsymbol{\mathbf{x}}}^*,\lambda,{\boldsymbol{\beta}})} = (\lambda e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}^*})^{-1}$. 2. $p(\lambda,{\boldsymbol{\beta}}|D_t) = \delta(\hat{\lambda} - \lambda)\delta(\hat{{\boldsymbol{\beta}}} - {\boldsymbol{\beta}})$ where $(\hat{\lambda},\hat{{\boldsymbol{\beta}}}) = \text{argmax}_{(\lambda,{\boldsymbol{\beta}})} p(\lambda,{\boldsymbol{\beta}}|D_t)$. The Dirac delta function $\delta(x)$ is loosely defined by $\delta(0)=\infty$ and is zero elsewhere. These approximations allow evaluation of the integrals (\[eq:energy\], \[eq:pred\]) and, additionally, it is computationally faster to obtain $(\hat{\lambda},\hat{{\boldsymbol{\beta}}})$ rather than numerically integrating (\[eq:energy\], \[eq:pred\]). Combining the above approximations we can write $\hat{t} = (\hat{\lambda}e^{\hat{{\boldsymbol{\beta}}}\cdot{\boldsymbol{\mathbf{x}}}^*})^{-1}$ and obtain $$\begin{aligned} \hat{H}({\boldsymbol{\mathbf{x}}}^*|D_t) &= \hat{h}(D_t\cup\{{\boldsymbol{\mathbf{x}}}^*,\hat{t}\})\label{eq:approxE}\\ \hat{h}(D_t) &= -\left<\log q(\lambda)\right>_{q(\lambda)} - \left<\log q({\boldsymbol{\beta}})\right>_{q({\boldsymbol{\beta}})}\label{eq:approx}.\end{aligned}$$ These can be substituted into (\[eq:objective\]) to obtain an approximated objective function $\hat{E}({\boldsymbol{\mathbf{x}}}^*|D_t)$. Evaluation of these expressions require numerical optimisation of (\[eq:posterior\]) and (\[eq:kl\]) in order to evaluate, but this is computationally feasible. Note that estimates of $\lambda$ and ${\boldsymbol{\beta}}$ could be unstable at the early stages of the trial when few patients have been recruited. In this case, one could implement a ‘burn in’ phase where selective recruitment only begins after a certain number of patients have been recruited. Obtaining a recruitment and allocation probability {#sec:palloc} -------------------------------------------------- Once a candidate patient presents with covariates ${\boldsymbol{\mathbf{x}}}^*$ we would like to define a recruitment probability $\rho({\boldsymbol{\mathbf{x}}}^*|D_t)$. In general, we can write ${\boldsymbol{\mathbf{x}}}^* = [{\boldsymbol{\mathbf{y}}}^*,{\boldsymbol{\mathbf{z}}}]$ where ${\boldsymbol{\mathbf{y}}}^*$ are clinical covariates or biomarkers and ${\boldsymbol{\mathbf{z}}}$ indicates the allocated treatment arm. Suppose there are $K$ arms in total and ${\boldsymbol{\mathbf{z}}}\in \{{\boldsymbol{\mathbf{z}}}_1,\ldots,{\boldsymbol{\mathbf{z}}}_K\}$ where ${\boldsymbol{\mathbf{z}}}_{k}$ indicates allocation to arm $k$. The first step is to define the allocation probability to treatment arm $k$ as $$p(k|{\boldsymbol{\mathbf{x}}}^*,D_t) = \frac{\hat{E}({\boldsymbol{\mathbf{y}}}^*,{\boldsymbol{\mathbf{z}}}_{k}|D_t)}{\sum_{j=1}^K \hat{E}({\boldsymbol{\mathbf{y}}}^*,{\boldsymbol{\mathbf{z}}}_{j}|D_t)}\quad\text{for $k=1,\ldots,K$.} \label{eq:biased}$$ A treatment arm is chosen at random according to this distribution and is denoted by ${\boldsymbol{\mathbf{z}}}^*$. Secondly, we define the *ideal* candidate as ${\boldsymbol{\mathbf{y}}}_{I} = \text{argmax}_{{\boldsymbol{\mathbf{y}}}} \hat{E}({\boldsymbol{\mathbf{y}}},{\boldsymbol{\mathbf{z}}}^*|D_t)$. The ideal candidate would give us the greatest reduction in expected entropy. A recruitment probability is given by $$\rho({\boldsymbol{\mathbf{x}}}^*|D_t) = f_0\left(\frac{\hat{E}({\boldsymbol{\mathbf{y}}}^*,{\boldsymbol{\mathbf{z}}}^*|D_t)}{\hat{E}({\boldsymbol{\mathbf{y}}}_I,{\boldsymbol{\mathbf{z}}}^*|D_t)}\right) \label{eq:rho}$$ where $f_0$ is some function that remains to be specified. Since the argument of $f_0$ must lie in the interval $[0,1]$ we can choose $f_0$ to be the identity function in which case the closer the candidate is to the ideal patient the higher the probability of recruitment. Alternatively, we can choose $f_0(s) = \theta(s - p_0)$ for a specified threshold $p_0$. The step function $\theta(s) = 0$ if $s\leq0$ and $\theta(s)=1$ otherwise. This results in deterministic recruitment. A more general option is $f_0(s) = (1+\text{tanh}(s/\beta_0-p_0))/2$ which is equivalent to deterministic recruitment when $\beta_0\to0$. This allows the practitioner to implement a desired level of stringency in the recruitment process. The German Breast Cancer Dataset {#sec:results:gbcs} ================================ We applied our method to data obtained from the German Breast Cancer Study (GBCS) described in . Our goal is to infer the parameters for a single covariate in order to assess how our adaptive protocol performs. The data consist of time-to-event outcomes for 686 patients recruited between July 1984 and December 1989. There are eight covariates in total. We decided to use tumour size (mm) for a univariate analysis because a good spread (1st quartile $= 20$ mm, median = $25$ mm, 3rd quartile $=35$ mm) would make it suitable for filtering patients according to the covariate. Importantly, the dataset also contains the date at which each patient is diagnosed with primary node positive breast cancer so we can easily calculate the waiting-time between patients. This allows us to effectively ‘re-run’ the trial. The primary event was recurrence. ![Plot of the posterior entropy (\[eq:approx\]) for the RCT and ACT as a function of time. The vertical ticks indicate times at which a patient was recruited. The sharp drop at $\approx0.75$ years corresponds to the first primary event occurring.[]{data-label="fig:entropy"}](fig4.pdf) To assess the information-adaptive design we decided to recruit a total of $N_{T}=100$ patients. We used deterministic recruitment with a cutoff of $p_0 = 0.5$. The trial was terminated after 10 years. We compared this to a randomised clinical trial (RCT) in which the first 100 patients are recruited. The same proportional hazards model as Section \[sec:ph\] was used to analyse the RCT. The covariate values were median-centred and rescaled by 25 mm. The population density was assumed constant. We impose a uniform prior between $\pm1$ for the ideal covariate $x_{I}$. Hyperparameters were set to $(\kappa_0,\chi_0,\alpha_0^2)=(3,1,4)$. $N_{total}$ $N_{reject}$ $t_R$ $\lambda$ $\beta$ entropy ------ ------------- -------------- ------- ----------- ---------------------------------------- --------- Full 686 0 67 0.13 0.36 (0.19,0.52), $p=6.1\times10^{-6}$ -4.54 ACT 100 278 31 0.11 0.44 (0.21,0.66), $p=4.2\times10^{-5}$ -3.49 RCT 100 0 11 0.14 0.11 (-0.27,0.48), $p=0.29$ -2.83 : Inferred parameters and entropies of the full GBCS dataset (Full), the adaptive clinical trial (ACT), and the randomised clinical trial (RCT). In brackets are 95 percent confidence intervals and $p$ is corresponding the p-value. $N_{total}$ is the total number of recruits, $N_{reject}$ is the number of rejected candidates, and $t_R$ is the recruitment time in months.[]{data-label="tab:gbcs"} It took approximately 1 year to recruit 100 patients onto the RCT. The adaptive clinical trial (ACT) took approximately 2.5 years, during which a total of 278 patients were rejected. In Figure \[fig:entropy\] the posterior entropies for both the ACT and RCT are plotted. Initially the entropies are largely determined by the priors over $\lambda$ and $\beta$ but quickly drop as patients are recruited, although not monotonically. In the first 2.5 years of the trial the RCT has a lower entropy which is presumably due to the fact that more patients have been recruited compared to the ACT. Towards the end of the trial the ACT has a lower entropy due to a more informative cohort. Both entropies continue to decrease after recruitment has finished as more events are observed. Table \[tab:gbcs\] shows the inferred model parameters (evaluated after 10 years) from the original dataset, the ACT, and the RCT. The ACT results in a significant non-zero value for $\beta$ that is close to the value obtained using the full dataset (with $N=686$). The RCT fails to infer any significant value. In order to gain some intuition for how the recruitment probabilities are determined we have plotted the expected entropy as a function of the covariate $x$ at various time points in Figure \[fig:energy\]. We note that the function tends to have one maximum and two minima at $x=\pm1$. This general shape is due to the nature of the proportional hazards model since extreme values of $x$ will diminish the space of plausible parameter values more so than values close to zero, and consequently are more informative. The dashed line is the entropy below which a candidate will be recruited. In (a) the trial has started at $t=0$ with two patients. There is a strong preference for individuals towards $\pm 1$. The next candidate (at $t=34$ days) had $x^*=-0.52$ and so was recruited. In (b), some patients with covariate values $>1$ have been recruited and this encourages recruitment of negative covariate values. At $t=267$ days no primary events have occurred. In (c), after $t=268$ days the first primary event occurs for a patient with a positive covariate value. This additional piece of information further increases the benefit of recruiting negative covariate values over positive ones. Note that the vertical scale changes. This illustrates that the recruitment probability changes dynamically, and depends on the observed events and covariate values of the existing cohort. We conclude that in general we gain more information from covariate values that have been under-sampled or values where few primary events have occurred. Individuals with covariates values far from zero will have the greatest reduction in expected entropy. This is because these terms will dominate the data likelihood in a proportional hazards model. Consequently, the covariate distribution in the ACT can differ considerably from the population distribution. Figure \[fig:density\] shows the empirical covariate distributions for the original dataset and both trials. Due to the shape of the expected entropy function (see Figure \[fig:energy\]) patients towards $\pm 1$ were more likely to be recruited in the ACT. Consequently, almost no patients with $x\approx 0$ were recruited. The RCT density resembles the density of the full dataset. ![The expected entropy (\[eq:approxE\]) as a function of $x$ at various times during the ACT.[]{data-label="fig:energy"}](figB.pdf) Numerical simulation studies {#sec:results:num} ============================ Here we consider a scenario where the covariates consist of a two-dimensional biomarker ${\boldsymbol{\mathbf{y}}}_i =(y_{i1},y_{i2})$ and patients are given one of three treatments denoted by ${\boldsymbol{\mathbf{z}}}_i = (z_{i1},z_{i2},z_{i3})$. A patient given treatment one would have ${\boldsymbol{\mathbf{z}}}_i = (1,0,0)$, treatment two would have ${\boldsymbol{\mathbf{z}}}_i=(0,1,0)$, and so forth. We are interested in whether there is any interaction between the biomarker and treatments, i.e. is the biomarker predictive. A proportional hazards model with interaction terms is assumed: $$h(t|{\boldsymbol{\mathbf{y}}}_i,{\boldsymbol{\mathbf{z}}}_i,\lambda,{\boldsymbol{\beta}}) = \lambda e^{\beta_1y_{i1}z_{i1} + \beta_2y_{i1}z_{i2} + \beta_3y_{i1}z_{i3} + \beta_4y_{i2}z_{i1} + \beta_5y_{i2}z_{i2} + \beta_6y_{i2}z_{i3}}. \label{eq:interaction}$$ This gives a total of six regression coefficients and the baseline hazard $\lambda$ to be inferred. In all simulations we compared an adaptive trial to a randomised one. To simulate survival data we generate a random vector ${\boldsymbol{\mathbf{y}}}=(y_1,y_2)$ where $y_i\sim \text{uniform}(-1,+1)$ or $y_i\sim \mathcal{N}(0,0.5)$ for $i=1,2$. A treatment arm ${\boldsymbol{\mathbf{z}}}$ is chosen (either randomly or according to (\[eq:biased\])). A random number $w\sim \text{uniform}(0,1)$ is generated, and an event time is given by the inverse of the cumulative distribution $t = - e^{-{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}}\log(1-w)/\lambda$ where ${\boldsymbol{\mathbf{x}}}\in\mathbb{R}^6$ contains the same product terms between ${\boldsymbol{\mathbf{y}}}$ and ${\boldsymbol{\mathbf{z}}}$ as (\[eq:interaction\]). Patients are censored at random with probability $p_{c}\in[0,1]$. If an individual is censored then the time-to-censoring is drawn from a uniform density between 0 and $t$. The first patient to be generated is recruited onto both the ACT and RCT. The waiting time until the next patient is drawn from an exponential density with rate parameter $\xi$. Hyperparameters were set to $(\kappa_0,\chi_0,\alpha_0^2)=(3,1,4)$. ![Kernel smoothed empirical covariate densities (Gaussian kernel, bandwidth $=0.2$) for (a) the full GBCS dataset, (b) the ACT, and (c) the RCT.[]{data-label="fig:density"}](figA.pdf) $\beta_1$ $\beta_2$ $\beta_3$ $\beta_4$ $\beta_5$ $\beta_6$ $\lambda$ ---------------- ----------- ----------- ----------- ----------- ----------- ----------- ----------- ACT (Uniform) 0.348 0.374 0.361 0.384 0.418 0.352 0.00080 RCT (Uniform) 0.364 0.347 0.401 0.389 0.396 0.384 0.00084 ACT (Gaussian) 0.499 0.5120 0.487 0.438 0.445 0.430 0.00085 RCT (Gaussian) 0.470 0.494 0.504 0.471 0.518 0.435 0.00084 : Mean square error between inferred and ‘true’ model parameters over 500 simulations. Comparison between both random and adaptive trials without selective recruitment and uniform and Gaussian distributed covariates.[]{data-label="tab:ap1"} Adaptive allocation without selective recruitment ------------------------------------------------- In these simulations all patients were recruited. A total of $N=50$ patients were recruited onto both trials. The trial was terminated after $t=100$ arbitrary units of time. The rate parameter for waiting times was $\xi=6$, and $p_{c}=0.5$. Model parameters were set to ${\boldsymbol{\beta}}=(0.8,-0.5,1.1,-0.7,0.6,0.1)$ and $\lambda=0.1$. In the ACT the expected entropy was used to determine which treatment arm each individual was allocated to as described in Section \[sec:palloc\]. In the RCT patients were allocated to one of the three arms at random. A total of 500 simulations were run. We computed the mean square error between the inferred model parameters and the ‘true’ values used to generate the data. As shown in Table \[tab:ap1\] we found essentially no difference between the randomised and adaptive trials for either uniformly or Gaussian distributed covariates. We found that the entropy at the end of the ACTs with uniform covariates was on average slightly lower than the RCTs (2.14 and 2.20 respectively), although the difference was statistically significant (p-value $0.017$ with a one-sided paired t-test). For Gaussian distributed covariates the difference in entropies was insignificant. We also performed a chi-squared test to see if the allocation proportions of patients across arms differed from a uniform distribution. Each simulated trial was tested and we found no p-values less than 0.05 for either uniform or Gaussian distributed covariates. Since the chi-squared test was repeated for each trial the p-values were corrected for multiple hypothesis testing by controlling the false discovery rate (using the method of @benjamini1995controlling) with the ‘p.adjust’ R function. Adaptive allocation and recruitment ----------------------------------- In these simulations the same parameters as above were used but patients were recruited onto the ACT selectively with a threshold of $p_0=0.66$. Over 500 simulations we found that the mean square error between the inferred and ‘true’ parameters was considerably lower in the ACTs than the RCTs as shown in Table \[tab:ap\]. For uniformly distributed covariates 48.9% of the inferred parameter values were significant (at 0.05) in the ACT compared to 39.2% in the RCTs. Furthermore, the mean entropy at the end of the ACTs was $0.93$, compared to $2.23$ in the RCTs. On average 140.7 (standard deviation 42.9) individuals are rejected. In the case of Gaussian distributed covariates the difference is more pronounced. 50.4% of parameters were significant in the ACT compared to 35.0% in the RCT. An average of 240.0 patients were rejected (standard deviation 61.9). Due to the Gaussian distribution there are more patients in the less informative region around zero. Therefore the number of rejections is higher and the benefit more substantial. We also explored the effect of the threshold $p_0$ on the trial results. When $p_0=0.33$ we found that the MSE (averaged over the six beta values) was 0.287 in the ACT compared to 0.372 in the RCT with 44.0% of inferred parameters reaching statistical significance in the ACT compared to 39.6% in the RCT. An average of 22.0 patients were rejected (standard deviation 6.45). When the threshold was increased to $p_0=0.90$ the MSE was 0.358 versus 0.363, and the proportion of significant parameters was 41.7% versus 39.3%, in the RCT and ACT respectively. On average 237.3 (standard deviation 86.5) patients were rejected. This suggests that setting the threshold too high can be counterproductive. $\beta_1$ $\beta_2$ $\beta_3$ $\beta_4$ $\beta_5$ $\beta_6$ $\lambda$ ---------------- ----------- ----------- ----------- ----------- ----------- ----------- ----------- ACT (uniform) 0.324 0.279 0.313 0.342 0.279 0.306 0.00079 RCT (uniform) 0.401 0.335 0.408 0.375 0.367 0.361 0.00081 ACT (Gaussian) 0.266 0.289 0.278 0.217 0.253 0.262 0.00085 RCT (Gaussian) 0.444 0.553 0.509 0.521 0.502 0.478 0.00082 : Mean square error between inferred and ‘true’ model parameters over 500 simulations. Comparison between random and adaptive trials with selective recruitment.[]{data-label="tab:ap"} Discussion {#sec:disc} ========== The practicality of our proposed design will depend on various economic and ethical considerations as well as the characteristics of each particular trial and the study population. For instance, if a covariate is relatively inexpensive to measure when compared to the costs of recruitment (treatment provision, follow-up, administration) then it may be sensible to selectively recruit informative patients. A large pool of patients can be inexpensively screened and then resources concentrated on those which are likely to provide the most information. In this case a selective recruitment design could result in significant cost reductions since fewer recruits are required overall. Clinical trials are not primarily intended to be therapeutic, but rather as a means to generate medical evidence. Recruited patients may be exposed to treatments that are ineffective (e.g. a placebo) or that are possibly even harmful. Our proposed design offers the possibility to conduct a trial using fewer patients than a traditional randomised design. This may be ethically attractive in some cases since ultimately fewer patients are offered treatment options with uncertain efficaciousness. In a selective recruitment design the decision to recruit and allocate a patient can also take into account the probability of a successful response to treatment (although this was outside the scope of this paper). Patients can be recruited and allocated in a manner that balances the statistical informativeness of a decision against the potential benefit or harm to that individual. The decision making process must balance individual and collective benefits. Maximising statistical information offers a collective benefit to all patients outside the trial (both current and future) who could benefit from the trial findings. Naturally this must be offset by what is best for the trial participants. What our proposed design offers the practitioner is a framework to balance individual versus collective ethical considerations. Selective recruitment designs suffer from a number of drawbacks, one of which is longer recruitment times. If the patient accrual rate is low it may render the overall recruitment period unfeasible. Selective recruitment designs are therefore only appropriate in situations where patients accrue relatively quickly or where longer recruitment periods are an acceptable compromise. One of the consequences of a proportional hazards model is that the most informative patients tend to have extreme values of covariates. As a result the distribution of recruited patients may differ from the population distribution which might make it difficult to generalise results from the trial to the general population. Thus, some generalisability is sacrificed in return for greater statistical power. If this was deemed undesirable one could introduce a sufficient level of random sampling in addition to preferential accrual of informative patients. Each candidate patient has a minimum probability of recruitment with informative patients having a higher probability. Thus, selective recruitment need not be an all or nothing process; it can be used to enrich the trial with informative patients to a desired degree. Finally, in the case of model misspecification undesirable biases may be introduced into the dataset because the model choice influences the covariate distribution considerably. An additional limitation is that it is not yet clear how to estimate the sample size required for a certain level of statistical power — a calculation that is typically used when planning new trials. In summary, our novel information-adaptive selective recruitment clinical trial design will reject non-informative patients. Individuals who are more likely to clarify the values of our model parameters are more likely to be recruited. We have demonstrated with both experimental and simulated data the feasibility of our approach. Statistically significant inferences can be achieved using fewer patients with a selective recruitment design than a randomised trial, although we found that treatment arm allocation using an entropy based measure (without selective recruitment) did not offer any improvement over a randomised design. Such a design may offer a more economical or ethically attractive route to discover the relationship between biomarkers, treatments, and survival outcomes. It will be interesting to extend this work beyond the proportional hazards assumption to more complex survival models. Incorporation of response-adaptive protocols offer another promising extension. Throughout this work we have assumed a uniform population density. In the case of a non-uniform density it may be desirable to incorporate this into the definition of an ideal candidate such that an ideal candidate is both informative and likely to be observed. This will require further investigation. Further extensions of the model could include alternative outcomes such as binary or continuous measurements. Derivation of the Kullback-Leibler divergence {#app:details} ============================================= The first two terms of the Kullback-Leibler divergence (\[eq:kl\]) in Section \[sec:var\] are simply minus the entropies of the variational distributions. These are $\left<\log q(\lambda)\right>_{q(\lambda)} = -(1/2+\log(2\pi\sigma_1^2)/2+\mu_1)$ and $\left<\log q({\boldsymbol{\beta}})\right>_{q({\boldsymbol{\beta}})}= -\sum_{\nu=1}^d\log(2\pi e \sigma_{0\nu}^2)/2$. The third term from (\[eq:kl\]) is $$\begin{aligned} -N^1_t\left<\log\lambda\right>_{q(\lambda)} - {\boldsymbol{\Phi}}_t\cdot\left<{\boldsymbol{\beta}}\right>_{q({\boldsymbol{\beta}})}&+\left<\lambda\right>_{q(\lambda)}\sum_{i=1}^{N_t}t_i\left<e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}_i}\right>_{q({\boldsymbol{\beta}})}\nonumber\\ & - \left<\log p(\lambda|\kappa_0,\chi_0)\right>_{q(\lambda)} - \left<\log p({\boldsymbol{\beta}}|\alpha_0^2)\right>_{q({\boldsymbol{\beta}})}\end{aligned}$$ where $N_t^1$ is the number of non-censored events up until time $t$ and ${\boldsymbol{\Phi}}_t = \sum_{i:\Delta_i=1}{\boldsymbol{\mathbf{x}}}_i$. It is straightforward to show $\left<\log\lambda\right>_{q(\lambda)} = \mu_1$, $\left<\lambda\right>_{q(\lambda)} = e^{\mu_1+\sigma^2_1/2}$ and $\left<{\boldsymbol{\beta}}\right>_{q({\boldsymbol{\beta}})} = {\boldsymbol{\mu}}_0$. The following result is needed [@Coolen2005 Appendix D]: $$\int\text{d}{\boldsymbol{\mathbf{z}}}\, \frac{e^{-\frac{1}{2}({\boldsymbol{\mathbf{z}}}-{\boldsymbol{\mu}})\cdot A^{-1}({\boldsymbol{\mathbf{z}}}-{\boldsymbol{\mu}}) + {\boldsymbol{\mathbf{b}}}\cdot{\boldsymbol{\mathbf{z}}}}}{(2\pi)^{d/2}|A|^{1/2}} = e^{{\boldsymbol{\mu}}\cdot{\boldsymbol{\mathbf{b}}}+ \frac{1}{2}{\boldsymbol{\mathbf{b}}}\cdot A{\boldsymbol{\mathbf{b}}}} \label{eq:mgf}$$ from which it follows $\left<e^{{\boldsymbol{\beta}}\cdot{\boldsymbol{\mathbf{x}}}_i}\right>_{q({\boldsymbol{\beta}})} = e^{{\boldsymbol{\mu}}_0\cdot{\boldsymbol{\mathbf{x}}}_i + \frac{1}{2}{\boldsymbol{\mathbf{x}}}_i\cdot{\boldsymbol{\Sigma}}_0{\boldsymbol{\mathbf{x}}}_i}$. Note that (\[eq:mgf\]) also defines the moment generating function for a multivariate normal distribution with mean ${\boldsymbol{\mu}}$ and covariance matrix $A$. The terms relating to the priors are $\left<\log p({\boldsymbol{\beta}}|\alpha_0^2)\right>_{q({\boldsymbol{\beta}})} =-\sum_{\nu}(\sigma^2_{0\nu} + [{\boldsymbol{\mu}}_0]_{\nu}^2)/2\alpha_0^2$ and $\left<\log p(\lambda|\kappa_0,\chi_0)\right>_{q(\lambda)} = (\kappa_0-1)\left<\log\lambda\right>_{q(\lambda)} - \chi_0^{-1}\left<\lambda\right>_{q(\lambda)}$ where $[{\boldsymbol{\mu}}_0]_{\nu}$ denotes the $\nu$th component of ${\boldsymbol{\mu}}_0$.

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Phenotypic and genotypic characterization of clinical and intestinal enterococci isolated from inpatients and outpatients in two Brazilian hospitals. The phenotypic and genotypic characteristics of clinical and intestinal enterococcal isolates recovered from inpatients and outpatients of two Brazilian hospitals, located in Niterói city, Rio de Janeiro, Brazil, were compared. A total of 601 strains were studied, including 253 isolated from different clinical sources and 348 intestinal strains (205 isolated from inpatients and 143 from outpatients) recovered from fecal specimens. Isolates were identified by using conventional physiological tests and evaluated for high-level resistance to aminoglycosides (HLR-A) and resistance to vancomycin and ampicillin by the agar screening technique. Susceptibility to several antimicrobial agents was evaluated by the disk diffusion method. The genetic diversity of Enterococcus faecalis strains presenting HLR-A was assessed by pulsed-field gel electrophoresis of chromosomal DNA after SmaI digestion. E. faecalis was the most frequent species among clinical isolates (90.1%) and intestinal strains from inpatients (53.6%). E. casseliflavus was the prevalent species among intestinal isolates from outpatients (35.0%). Clinical isolates were shown to be resistant to erythromycin (53.0%), tetracycline (52.2%), ciprofloxacin (36.4%), gentamicin (36.4%), streptomycin (30.4%), chloramphenicol (34.4%), norfloxacin (32.0%), imipenem (3.2%), and ampicillin (2.8%). Vancomycin resistance was only detected in intrinsic vancomycin-resistant enterococcal species. The overall prevalence of HLR-A was 52.2% among clinical isolates and 40.5% among intestinal strains. However, HLR-A was significantly more frequent among intestinal strains obtained from inpatients (56.6%) than among strains from outpatients (17.5%). Three major clonal groups were found among E. faecalis strains exhibiting HLR-GE or HLR-GE/ST (clonal groups GE-A and GE-B), and strains exhibiting HLR-ST (clonal group ST-A). HLR-A, particularly HLR-GE, was most frequently associated with enterococcal strains of nosocomial origin. Isolates included in the major clonal groups were recovered from clinical and intestinal sources from patients in both hospitals, indicating both intrahospital and interhospital spread of strains.

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The present invention relates generally to series progressive divider valves. More particularly, the present invention relates to fittings for plugging outlet ports such that fluid can be directed to the next outlet port in the series progression. Series progressive divider valves have long-existed in the art and comprise a mechanism for dividing a single, steady input of pressurized fluid into multiple, distributed bursts of fluid. Thus, fluid is delivered to the valve body at a single inlet port and delivered to multiple discrete outlet ports through cyclic operation of an array of pistons or spools under pressure from the fluid. The valve output cycles continuously through the outlet ports in a scheduled progression based on movement of the array of pistons. For example, conventional series progressive divider valves include an array of pistons in which the central axes of all the pistons are arranged in a single plane. Outlets for each end of the piston are typically arranged in a plane parallel to the plane of the pistons. The outlets are connected to the pistons through an elaborate system of portings machined into the valve body. The pistons reciprocate within bores of the valve body enclosed by end caps. The pistons themselves include a pair of axially spaced undercuts such that each piston forms three lobes. As such, when a piston is inserted into a bore and enclosed by end caps, four pressure chambers are formed: one end chamber at each end of the piston and two internal chambers within the piston. Each end chamber is connected to an internal chamber of the next piston in the progression through porting extending through the valve body. Additionally, each internal chamber is connected to an outlet of the valve through the use of separate porting. Thus, a four piston valve includes eight outlets. High pressure inlet porting connects each piston bore and, depending on the position of each piston, one of the internal chambers for each piston. All connections and outlets are made on the same side of the valve body and at the same ends of the pistons, except, however, end chambers of a “first” piston are connected to internal chambers of a “last” piston such that the pistons can reverse direction and the series progression can continue ad infinitum. Operation of a typical series progressive divider valve is explained with reference to drawings from the prior art, specifically U.S. Pat. No. 4,312,425 to Snow et al., which shows a simplified piston and outlet configuration. FIG. 1 shows a perspective view of a typical series progressive divider valve having valve body 10 formed from a plurality of block bodies 10A-10H. FIG. 2 shows a schematic of valve body 10 including pistons 12A, 12B and 12C. FIG. 1 and FIG. 2 are discussed concurrently. As shown, each of pistons 12A-12C includes three lobes, designated as 14A, 14B, 14C, 16A, 16B, 16C, 18A, 18B and 18C, respectively. Lobes 14A-18C produce undercuts 20A, 20B, 20C, 22A, 22B and 22C, respectively. Pistons 12A, 12B and 12C reciprocate in bores 24A, 24B and 24C, respectively, which form end chambers 26A, 26B, 26C, 28A, 28B and 28C, respectively. Additionally, undercuts 20A-22C form internal chambers 30A, 30B, 30C, 32A, 32B and 32C. Each of undercuts 20A-22C, and the chamber formed thereby, is fluidly connected to one of valve outlets 38A-38F and another undercut via porting machined into valve body 10. Specifically, internal pumping chamber 30A is fluidly connected to end chamber 28C via porting 36A and to outlet 38A via porting 40A. Internal pumping chamber 30B is fluidly connected to end chamber 26A via porting 36B and to outlet 38B via porting 40B. Internal pumping chamber 30C is fluidly connected to end chamber 26B via porting 36C and to outlet 38C via porting 40C. Internal pumping chamber 32A is fluidly connected to end chamber 26C via porting 36D and to outlet 38D via porting 40D. Internal pumping chamber 32B is fluidly connected to end chamber 28A via porting 36E and to outlet 38E via porting 40E. Internal pumping chamber 32C is fluidly connected to end chamber 28B via porting 36F and to outlet 38F via porting 40F. High pressure porting 42 distributes high pressure fluid from inlet 44 to bores 24A-24C. High pressure porting 42 fluidly connects inlet 44 to internal chambers 30A-32C, depending on the position of lobes 16A-16C. High pressure fluid is always provided directly to one side of each of lobes 16A-16C depending on the position of each of pistons 12A-12C. As shown, high pressure fluid is provided to internal chambers 32A, 30B and 30C. As such, high pressure fluid is also provided to end chambers 26C, 26A and 26B, via porting 36D, 36B, 36C, respectively. In the last piston movement before the configuration shown in FIG. 2, low pressure fluid has been dispensed from port 38F via movement of piston 12B downward through porting 36F and 40F. Subsequently, as shown in FIG. 2, high pressure fluid is provided to chambers 26B and 26C. High pressure fluid in chambers 26B and 26C does not produce movement of pistons 12B and 12C because lobes 18B and 18C are already engaged with the end caps of bores 24B and 24C. High pressure fluid in chamber 26A will, however, produce downward movement of piston 12A as end chamber 28A discharges low pressure fluid. Low pressure fluid in end chamber 28A, through porting 36E, displaces fluid in internal chamber 32B out of outlet 38E through porting 40E. Such displacement of pistons 12A-12C is repeated so long as high pressure fluid is provided to inlet 44, with porting 36D and 36A connecting internal chambers and end chambers on opposite ends of the pistons to permit reversing of the axial piston positions. For example, piston 12C moves downward pushing fluid through outlet 38A, piston 12B then moves downward pushing fluid through outlet 38F, piston 12A then moves downward pushing fluid through outlet 38E, then piston 12C moves upward pushing fluid through outlet 38D, then piston 12B moves upward pushing fluid through outlet 38C and finally piston 12A moves upward pushing fluid through outlet 38B. As mentioned, in order to achieve such cyclic movement, valve body 10 is provided with an elaborate system of three dimensional porting. Such porting is produced using a series of machining operations, particularly drilling, in a plurality of rectangular blocks. For example, valve body 10 is produced from blocks 10A-10H as shown in FIG. 1. Blocks 10A and 10E comprise “inlet” and “end” blocks with porting necessary to route fluid between pistons at the end of the array. Intermediate blocks 10B-10D are identical to each other and include piston bores 24A-24C. Intermediate blocks 10E-10H are identical to each other and include outlets 38A-38F. Intermediate blocks 10B-10D are paired with intermediate blocks 10E-10H to form a piston and outlet combination. In order to change the number of pistons and outlet ports one pair of intermediate blocks can be removed. However, such an operation requires tedious and time consuming disassembly and reassembly of the blocks, such as by removal and replacement of screws 46A-46I. Such assembly intricacies are further detailed are described in the aforementioned U.S. Pat. No. 4,312,425 to Snow et al. The use of a plurality of separate intermediate blocks reduces or eliminates the need for unnecessary “open ended” drilling operations. These drilling operations are intended to connect other passages, but are not intended to produce a passage that opens to the exterior of the valve body. However, due to manufacturing limitations the drilling operations are necessary and the open end must be plugged. For example, two parallel ports may need to be connected by drilling a perpendicular port. The perpendicular port does not, however, need to be opened to the exterior of the valve block. Such ports have typically been closed off using steel balls welded in place, as is described in U.S. Pat. No. 3,467,222 to Gruber. These methods thus require additional manufacturing steps and additionally introduce potential leak points and stress points into the system. In other configurations of prior art series progressive valves, outlet ports 38A-38F can be connected to each other through cross-porting and plugging. In particular, outlet 36C can be ported to connect with outlets 38A and 38B. Likewise, outlet 38F can be ported to connect with outlets 38E and 38D. When configured as such, outlet 38B can be plugged to direct what would be its discharge into outlet 38C so that outlet 38C will receive a double shot of fluid. Additionally, outlet 38C can be plugged so outlet 38A can be configured to receive a triple shot of fluid. However, in conventional series progressive valves using intermediate blocks, outlet 38A cannot be plugged because no porting is provided between outlets 38A and 38B due to the complexity of the required porting that cannot be introduced into the modular block design of blocks 10A-10H. In other words, the required porting would result in each intermediate block having a unique configuration. As such, outlet 38A becomes a “last stop” outlet that must be permitted to allow fluid from valve body 10 because there is not another outlet to which fluid can be routed. As such if outlets 38A, 38B and 38C were plugged, operation of valve body 10 would seize up. Outlet 38D also becomes a “last stop” outlet for the same reason. Cross-porting requires blocking of outlet ports with fittings or plugs from which it is desired to prevent fluid flow. Such cross-port fittings are shown in the Quicklub® Progressive Divider Valves brochure for SSV & SSVM Series valves commercially available from Lincoln Industrial, an SKF company. These fittings, however, require the use of several different plug and ferrule combinations. Other methods of output combining involve force fitting brass plugs into the outlets. These plugs, however, wear after repeated use and become ineffective.

Andrew Robathan Andrew Robert George Robathan, Baron Robathan, (born 17 July 1951) is a British Conservative politician, who served as Member of Parliament (MP) for South Leicestershire (previously Blaby) in Leicestershire as well as a government minister. In September 2014 Robathan announced he would stand down at the 2015 general election, and was nominated for a life peerage in 2015 Dissolution Honours, being created Baron Robathan, of Poultney in the County of Leicestershire, on 13 October 2015. Early life Robathan was born on 17 July 1951. He was educated at Merchant Taylors' School, an all-boys public school in Northwood, London. He went up to read Modern History at Oriel College, University of Oxford, where he graduated with a Bachelor of Arts (BA) in 1973, later proceeding Master of Arts. Military career Robathan was commissioned into the Coldstream Guards as a Second Lieutenant (on probation) (University Candidate) on 6 October 1974. He was given the service number 498738. His commission was confirmed and he was given seniority as a Second Lieutenant from 6 April 1971. He was promoted to Lieutenant, back dated to 6 October 1974, with seniority from 6 April 1973. Having attended the Staff College, Camberley, he was promoted to Major on 30 September 1984. He served for a period of time with the Special Air Service (SAS), and retired on 27 August 1989 being appointed to the Reserve of Officers. He worked for BP from 1991–92, but volunteered to return to the Army between January and April 1991 during the First Gulf War, serving as Chief of Staff of the Prisoner of War Guard Force in Saudi Arabia, Iraq and Kuwait. Parliamentary career Robathan was elected to Hammersmith and Fulham Council in May 1990, defeating the then-Labour Mayor in the Eel Brook Ward. He resigned as a Councillor in late 1991 to fight the 1992 general election, elected as MP for Blaby in 1992 succeeding Nigel Lawson, the former Chancellor of the Exchequer. Blaby had a 37% majority in 1992, but it was considerably reduced by major boundary changes in 1997, creating a rise in Liberal Democrat support within the constituency. Following a public enquiry by the Electoral Commission and submissions from the public including Robathan, the Blaby constituency was reconfigured as South Leicestershire. Robathan served four years on the Defence Select Committee between 1997 and 2001; he was also Chairman of the All-Party Cycling Group, and Vice-Chairman of the All-Party Renewable and Sustainable Energy Group, whilst an MP. Robathan was John Redwood's Parliamentary Private Secretary to Iain Sproat, Minister for Sport, in the Major administration before returning to the backbenches when the Conservatives lost the 1997 general election. He returned to the front-bench as Trade and Industry Spokesman in 2002. In the 2001 Conservative Party leadership election, Robathan was a notable supporter of Michael Portillo. After six months on the backbenches, Robathan was appointed Commons Liaison to the Lords and then a Defence Spokesman in which capacity he fought the 2005 general election. In the 2005 Conservative leadership election, Robathan was one of the very first MPs to declare his support for David Cameron being rewarded with one of only five paid posts as Opposition Deputy Chief Whip. In May 2010, he was appointed Parliamentary Under Secretary of State at the Ministry of Defence with responsibility for Welfare and Veterans. In 2011, he was a member of the Special Select Committee set up to scrutinise the Bill that became the Armed Forces Act 2011. In December 2011, campaigners called for him to be sacked after he compared the medal claims of 66,500 veterans of the Arctic convoys of World War II to the proliferation of honours made by "authoritarian regimes" and "dictators": "One can look, for instance, at North Korean generals who are covered in medals or Gaddafi or Saddam Hussein... We have taken the view in this country, traditionally, that medals will only be awarded for campaigns that show risk and rigour." Some 3000 sailors died on the convoys - which Winston Churchill described as the "worst journey in the world." Portsmouth MPs described his behaviour as "shameful" and "sickening." In October 2012 Robathan was told to calm down by Lindsay Hoyle, the Deputy Speaker, after complaining about noise levels from the public gallery. Soldiers from the "historic" 2nd Battalion Royal Regiment of Fusiliers who face being disbanded were "politely applauding MPs who spoke up on their behalf". A former captain contrasted MoD advice that they went to Afghanistan and Iraq to help democracy with the reality of their own threatened ejection from Parliament. Expenses Robathan claimed the maximum second home entitlement of £24,006 in the period 2008–09 though he was not one of the 343 MPs required to repay money by the Legg Report. He was one of 177 MPs listed by The Daily Telegraph who employed family members. The Independent Parliamentary Standards Authority report published two years later, whilst tightening the rules, confirmed that a member may employ one relative subject to general conditions relating to expenses. Robathan's expenses were generally in the bottom half of all MPs. On 24 August 2009, he was quoted in The Times newspaper suggesting that MP's salaries be increased to £110,000. These comments were heavily frowned upon by his colleagues in Parliament and the media. On 4 November 2013, it was reported in The Leicester Mercury newspaper and on BBC Radio Leicester that Robathan had claimed £4,587 expenses to pay for energy bills for his second home. Personal life He married Rachael Maunder in December 1991 in Westminster. They have a son (born December 1996) and daughter (born July 1999). Rachael has been a councillor on Westminster City Council since 2010, representing Knightsbridge and Belgravia ward. Robathan speaks French and German, and has been admitted as a Freeman of the City of London. References External links Guardian Unlimited Politics - Ask Aristotle: Andrew Robathan MP TheyWorkForYou.com - Andrew Robathan MP Biography South Leicestershire Conservative Association BBC Politics page News items Leicestershire education in March 2003 Category:1951 births Category:Living people Category:People educated at Merchant Taylors' School, Northwood Category:Alumni of Oriel College, Oxford Category:Coldstream Guards officers Category:Special Air Service officers Category:British Army personnel of the Gulf War Category:Conservative Party (UK) MPs for English constituencies Category:UK MPs 1992–1997 Category:UK MPs 1997–2001 Category:UK MPs 2001–2005 Category:UK MPs 2005–2010 Category:UK MPs 2010–2015 Category:Members of the Privy Council of the United Kingdom Category:Conservative Party (UK) life peers Category:Peers nominated by David Cameron Category:Northern Ireland Office junior ministers

Posted 23 November 2016 - 07:46 PM Hello. Now I am sure I will get my head bitten off here for what I am about to say, but here goes! You deserve better from your daddy! All littles do! Aftercare is a forgotten aspect that may do not give to their partners. Now I don't need to know what's going on in your relationship nor do I want to. That is yours and his business. So let's look at the fact here. You lied. Bad girl, yes. He gave you punishment for this offense. You did the punishment. Good girl! Now your very first punishment (kinda harsh in my eyes but that's just me!). You did this without question. The reason for that is you know you messed up. Now the thing about your daddy is he didn't care to say or do anything after the fact. You stayed on Skype with him to see if he would come around. Well he did come around and asknyou to play with him. No I am not daddy bashing him here! We don't know why he did this nor do you. From the way you stated it he is more into the controlling and sexual aspects of your relationship than the actually being a daddy to you. This is just one instance, so I can not say that for sure. Honestly its none of ours business. This is something you need to sit down and think about. Look back over the month and see if he actually did anything for YOU! Cuddles, non sexual play, messaging you out of the blue to let you know he was thinking of you. If he is all that and a bag of chips then like the above said, he might be feeling guilty. You will never know unless you talk to him. Now be prepared to find Mr grumpy pants on the other end of the line as you hung up on him. If you encounter that, let him know why you did it. That you were hurt by his insensitivity, and you were crushed over it. If he doesn't want to hear that, then you have your answer in your face of what he is all about. Good luck and take care. Remember you have those that care about you here!

Fyi - Mike ---------------------- Forwarded by Michael Terraso/OTS/Enron on 03/14/2000 07:22 AM --------------------------- "Terry D. Boss" <tboss@INGAA.org> on 03/14/2000 09:56:15 AM To: "Dave Johnson (E-mail)" <David.L.Johnson@enron.com>, ""Mike Terraso (E-mail)" <mterras@enron.com>, ""Tilford Vik (E-mail)" <tvik@enron.com> cc: " "Max Brown (E-mail)" <mbrown@enron.com>, Subject: As Predicted, the USA Today Article on Pipelines 03/13/00- Updated 11:56 PM ET ? When pipelines are time bombs 2 million miles of them deliver potential catastrophe every day By Patrick McMahon, USA TODAY BELLINGHAM, Wash. - Last summer, an underground pipeline ruptured in a city park here and sent a torrent of gasoline along a wooded streambed toward two 10-year-olds playing with a barbecue lighter. After the 16-inch pipeline had been hemorrhaging for an hour and 34 minutes, an explosion sent a fireball racing through the park. A plume of smoke rose 30,000 feet. Authorities say the blast probably was triggered by a spark from the lighter. "There was a spark and the sky turned orange, the boys both told me afterward," says Frank King, whose son, Wade, was one of the boys in the park. Wade and his friend, Stephen Tsiorvas, were burned over 90% of their bodies. They died the next day. Also killed was 18-year-old Liam Wood, who had graduated from high school five days before. The college-bound Wood was fly-fishing in the park when he was overcome by fumes, fell into the creek and drowned. "To many people, the boys were considered heroes," says Mark Asmundson, the mayor of this seaside city north of Seattle. In sparking the explosion, the boys kept the most extensive damage confined to the 241-acre park, he says. "The river of gasoline was heading right for the center of the city." On Monday, the Senate Commerce Committee held a one-day hearing here, about five minutes from where the incident occurred, on legislation to improve safety and government oversight of pipelines and the companies that operate them. The hearing included gripping, sometimes tearful, testimony from the boys' parents as well as statements from state and federal officials. "My baby died because of inaction. His death was preventable," said Katherine Dalen, Stephen's mother. More than 2 million miles of iron, steel and plastic pipes - some as large as 5 feet in diameter - snake beneath the earth and deliver oil, gasoline, natural gas and potential disaster across America every day. They range from the trans-Alaskan pipeline to tiny pipes carrying natural gas to people's homes. Safer than gasoline trucks or ocean-going tankers by most measures, once-remote pipelines are prompting new worries as sprawling metropolitan areas grow into their paths. U.S. pipeline accidents have fluctuated in the past 10 years. In the 1990s, there were 3,917 liquid fuel spills and natural gas leaks, roughly one a day. The incidents, most involving local lines carrying natural gas, resulted in 201 deaths, 2,826 injuries and $778 million in property damage from 1990 through 1999. There also are environmental costs. In January, one of the nation's largest pipeline companies, Koch Industries of Wichita, Kan., paid a $30 million civil fine to settle Environmental Protection Agency water-pollution charges involving 300 oil spills from 1990 to 1997. The EPA said Koch failed to inspect its pipelines and waited for leaks before making repairs. Liam Wood's mother, Marlene Robinson, is focused on stronger regulations nationwide, including more and better inspections, and regional watchdog committees. "I don't have any children left to protect," she said in an interview and repeated at the hearing. "This didn't have to happen to Liam, and it doesn't have to happen to other people's children." Although the cause of the Bellingham incident remains under investigation, one factor might have been damage to the pipeline from a backhoe when a water line was being installed in 1995. But investigators also have many questions about the role of Olympic Pipeline Co., which operates a network of pipelines along a 299-mile corridor from north of Bellingham to Portland, Ore. Investigators for the National Transportation Safety Board (NTSB) want to know whether Olympic ignored warning signs and whether it could have acted more swiftly after the leak. But the inquiry has been delayed by a parallel investigation by the U.S. Attorney's Office. Several Olympic employees have declined to testify and have invoked their constitutional rights against self-incrimination. Pipeline owners and government regulators defend the safety of transporting two-thirds of the nation's fuels by pipeline, and they warn that more rules could boost energy prices. Critics say pipelines are too loosely regulated, not fully inspected and, in some cases, deteriorating. The U.S. Transportation Department oversees pipelines. Its Office of Pipeline Safety sets standards for design, operation, maintenance and emergency response. Richard Felder, who heads the office, testified at Monday's hearing. "Our goal is to prevent incidents like Bellingham from ever happening again," he said. In an earlier interview, he defended his office's performance and the industry: "It's a good record. It's the safest form of transporting fuel, far and away." The office has extensive rules on pipeline safety, including requirements for signs along the route and programs to alert potential diggers, but "there's no such thing as risk elimination," Felder says. "All you can do is manage it." But the chairman of the federal government's transportation safety watchdog agency is unimpressed with Felder's office and says it deserves a grade of F. "It's been the most frustrating area I've had to deal with as chairman," says Jim Hall, NTSB chairman since 1994. He says no agency has a worse record of responding to NTSB recommendations. In a recent speech, he said there's no indication that the Office of Pipeline Safety "is in charge or that its regulations, its inspections, its assets, its staffing and its spirit are adequate to the task." Felder takes issue with much of the criticism, but he endorses calls for more research into pipeline safety. He said his office is in the final stages of preparing new safety standards for heavily populated and environmentally sensitive areas. He emphasized that the leading cause of pipeline failure is "third-party damage" from road, utility and construction work. The agency has worked to enhance a system to alert pipeline companies of digging with one telephone call. Felder is critical of local authorities who ignore pipelines when making decisions on housing and commercial development. "Local planning has not kept people away from pipelines," he says. "It's astounding to me." Felder bristles at critics' suggestions that his agency is a tool of the oil and gas industry. "I can't agree with that one iota," he says. "We try to balance the safety issue and the economic issue." The Senate Commerce Committee will consider a bipartisan bill sponsored by Sens. Patty Murray, D-Wash., and Slade Gorton, R-Wash.; Rep. Jack Metcalf, the Republican who represents Bellingham; and Rep. Jay Inslee, a Democrat whose district includes an operating section of the pipeline. The bill would require periodic internal inspections of pipelines, boost federal spending on research, increase the number of federal pipeline inspectors nationwide - there now are 55 - and expand states' regulatory authority. On Monday, the Office of Pipeline Safety gave the state of Washington the temporary right to do more inspections. The Murray-Gorton bill also would require federal certification of pipeline workers and expand the public's right to know about spills and leaks. "I was totally shocked and amazed when this happened," Murray says. "You always assume that your neighborhood is safe and somebody has taken care of this." The Bellingham rupture not only dumped 278,000 gallons of gasoline into this city of 63,000 near the Canadian border but also unleashed civic furor at Olympic Pipeline. "It's hard for people to get interested in things they can't see. Out of sight is out of mind," Mayor Asmundson says. He has visited the boys' grieving families and lobbied Congress. The blast "has dominated my life since June 10." Pipeline operators portray accidents as isolated to escape national scrutiny, he says. "The whole focus of the industry is containment. Do whatever you have to do locally, but don't stir things up nationally." Last week, Murray released a report she requested from the Transportation Department's inspector general. It faults the Office of Pipeline Safety for ignoring enhanced safety requirements, including increased inspections inside pipes, in highly populated and environmentally sensitive areas as Congress required in 1992 and 1996. A U.S. General Accounting Office audit of operations is due in May. Also pending is a decision from Felder and the pipeline safety office on whether to restart the 39-mile section shut down since the Bellingham accident. Olympic Pipeline has repaired the line and is eager to reopen it. People living along the route and officials are pushing for more extensive testing of the still-operating section of the line south of Bellingham. Felder says he has not decided whether it should be shut down. Last week, Felder's office ordered a new round of testing along the entire line, but critics such as Inslee are not convinced it will be enough. The tests must be conducted on the closed section of the pipeline before a decision is made on reopening it. Contributing: Scott Hillkirk Confidential: INGAA Member Use only Terry D. Boss VP Environment Safety and Operations INGAA tboss@ingaa.org ?

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Q: Is [actionbarsherlock] still needed? Since everything you need to make an app support older versions is now included inside the support libraries, do we still need actionbarsherlock (and actionbarsherlock-map)?? A: Yes, it is still needed. Technology becoming obsolete does not magically disappear any of the existing installations, or the need to maintain them (and maybe port things there or from there). It might in time do so (Though don't hold your breath, that would be decidedly unhealthy, we still have cobol), but disappearing the tag would not help in removing content absolutely noone ever wants to look at ever again anyway.

Turkish President Tayyip Erdogan, accompanied by his wife Emine Erdogan, greets supporters during a rally to mark the 563rd anniversary of the conquest of the city by Ottoman Turks, in Istanbul, Turkey, May 29, 2016. REUTERS/Murad Sezer ISTANBUL (Reuters) - No Muslim family should engage in birth control or family planning, Turkish President Tayyip Erdogan said on Monday, calling again on pious Muslims to have more children. “We will multiply our descendants. They talk about population planning, birth control. No Muslim family can have such an approach,” he said in a speech in Istanbul broadcast live on television. “Nobody can interfere in God’s work. The first duty here belongs to mothers,” he said. Women’s groups and opposition politicians have criticized Erdogan, a devout Muslim for telling women how many children to have and dismissing the Western idea of gender equality. He has previously equated birth control with treason.

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Parts left over from the metal barn that Black Page Brewing leased out beside White Oak Bayou out a few years back are now lying in a heap next to a wooden skeleton that’s taken the demolished structure’s place. The deconstruction began last month according to neighbors who called 311 on August 31 to report that it was happening, potentially, they said, without the required permits. An inspector showed up the next day to check things out, one of several field trips the city would make to the planned brewpub’s digs at the end of Glen Park St. over the next few weeks in response to multiple follow-up calls from nearby residents. By the time a demo permit did show up last Friday, the site had already been tagged twice by city officials: first for the premature teardown, and once again — as shown below — for additional unpermitted work: Here’s the next target in Houston’s continuing warehouse-to-brewery turnover trend: 1504 Chapman St. A group of local brewers got their hands on the 6,283-sq.-ft. building — pictured above from the south — in April and a budding Facebook page now shows its address as the location of a venue they’re calling Local Group Brewing. It’s within the same general parish as St. Arnold’s recently-opened beer cathedral and existing brewery. They’re both less than half a mile away on the other side of the former Union Pacific brownfield pictured below, now giving rise to the complex of mixed-use buildings dubbed Hardy Yards: Houston’s City Planning Commission approved a variance yesterday permitting a developer that plans to build a 4-story apartment building on the corner highlighted above not to extend Dunlop St. through the site, as otherwise required. Instead, plans call for the street to end at the south side of the complex, where it’ll be bounded by a new, 8-ft. tall fence. The request first showed up on the commission’s agenda on April 26, at which time a couple of residents came forward to complain about the heavy traffic on nearby Karcher and Angelo streets — which northbound drivers use to avoid the light at the intersection of Link and Fulton. Extending Dunlop through the site, they argued, would clear up some of that congestion. But a 60-ft.-wide swath of road like that would run over the garage, parking lot, pool, and dumpster area the developer plans to build at the center of the complex, as shown in the site plan below: The transit-themed entryway Lennar Homes wants to build to its 39-lot development — dubbed Fulton Station — on the corner of Fulton and Cavalcade will get another shot at city approval when it goes before Houston’s planning commission this afternoon. Lennar’s new residential neighborhood hugs the Charisma Design Studios & Art Gallery Building, west of the southbound stop for METRO’s Red Line in the middle of Fulton St. The gated entrance would go at the foot of a private park Lennar has planned just across the street from the rail platform, on the parcel highlighted red in the map below: A show-stopping announcement posted on the Walter’s Downtown Facebook page yesterday brings sad news for thrashers, metal-heads, punks, and indie fans: the 18-year-old live music venue on the corner of Naylor and Vine streets plans to close down on February 4. Walter’s moved to its current location — the former classic car showroom, video production studio, car parts distribution center, and cabinet warehouse pictured above — in 2011. Before that, the club was located on Washington Ave, in a building just east of Thompson St. that’s since been transformed into the office of Carnegie Custom Homes. The photo below views the venue from its north side on Naylor back in 2014: There’s a new outdoor stairway zig-zagging its way up to the top of the Raven Tower’s above-ground spot at the northwest corner of the White Oak Music Hall complex. The elevated bar opened in January 2016 on North St., just off N. Main east of I-45, but shut down in May of that year after its developer decided to address accessibility issues through renovations. It only received a certificate of occupancy — allowing the venue to operate as a bar — in April of this year. Railings and landings are still missing from the new stairway. Inside the podium, an indoor stairway wraps around a central elevator shaft that rises from the base of the tower up to its peacock-blue penthouse. Here’s a view of the tower from beyond an adjacent concert venue, one of several surrounding the White Oak Music Hall’s main building: More new features are imagined for the center of Houston than just the new Green Loop highlighted in the just-released Plan Downtown proposal. There’s also a mysterious new Downtown island. Where did it come from? It’s the result of digging the long-whispered North Canal Channel Bypass, a re-linking of White Oak and Buffalo Bayous north of Downtown. Existing bends and narrow banks along the 2 bayous just east of Main St. restrict the flow of stormwater during flooding events. According to reports, engineering studies have estimated that cutting a straighter diversion channel to bypass the oxbow could reduce flooding Downtown by 3.5 ft. But digging a new canal while maintaining the existing path of the bayou would create an island out of the area just north of Commerce St. An imagined map of the area in Plan Downtown’s report (rotated so North is aimed down and to the right) shows what car and pedestrian bridges might link it to the mainland: “If you don’t know that’s a big dip,” reports a reader who scouted the scene of the impromptu lake formed over the weekend on the lawn of the Near Northside’s Hollywood Cemetery, “you don’t appreciate just how much water that is.” The cemetery lies between Little White Oak Bayou and I-45, along the northeast edge of N. Main St. The water level has lowered a bit since these photos were taken on Sunday. Do note the bouquet, presumably perched above one of the completely submerged gravestones, in the right foreground of the view above. The new Holiday Inn Express about to begin construction at 3401 N. Main St. in the Near Northside will have some consistently quiet neighbors and some occasionally very loud ones — with the steady drone of the adjacent North Fwy. available to somehow bridge the gap. The 1.44-acre site, where the Casa Grande Mexican Restaurant stood until it was torn down 2 years ago (and Stuarts Drive-In before it), sits across N. Main St. from the Hollywood Cemetery (yes, the same cemetery featured in Wes Anderson movie Rushmore). And it’s just a bit more than a quarter-mile up N. Main from the White Oak Music Hall complex, whose outdoor concert habit spurred nearby residents kept up late at night by the noise to file suit against the venue — and later, the city of Houston — for failing to follow (and enforce) local sound ordinances. Late last month, crews removed the concrete paving left behind after the Casa Grande demolition (see photos above). Just this week, a city permit was granted for a 58,929-sq.-ft., 95-room Holiday Inn Express on the site — up 10 rooms from the 85 promised a couple of years ago, when the developers submitted these drawings as part of an application for a variance that would allow them not to have to extend or widen Norma St., on the north end of the lot: Currently listed for an undisclosed amount on CBRE’s website: a 10.69-acre chunk of the former Union Pacific railyard brownfield property previously sketched up for future conversion to the Hardy Yards mixed-use development. The section up for grabs appears to snuggle up to the west against a piece of land owned by Metro, whose Burnett Transit Center and light-rail Red Line are elevated above that semi-catching segment of N. Main St. tunnel; the parcel extends east to the new-ish segments of Fulton and Leona St., likely not too far from the spot where that rail car full of lithium batteriesblew up back in April. On the other side of the site, meanwhile, the Residences at Hardy Yardsapartments are under construction, per photos from the Zieben Group published back in May: Not to be outdone by last week’s midday plug-up of the Alfred Hernandez Tunnel beneath the railroad tracks and the Burnett TC Red Line stop, another semi making its way through the passage got lodged in the tunnel late this morning — getting torn open end-to-end in the process. But that’s not even the first truck stuckage incident at the underpass in the last 24 hours, according to a reader who’s had both a camera and a Twitter account trained on the recently retooled intersection for at least the last few months. The reader tells Swamplot that another truck got stuck briefly last night, and that it happens about 6 times a week: “Our camera system auto-wakes when it hears something beyond a certain threshold; most drive away, presumably nervous[ly] on their way to have a talk with the boss.” Some work on the tunnel has been on the city’s docket this spring, and was approved at a mid-April meeting; that’s likely to start around the end of the month. The light at the end of the N. Main St. tunnel beneath the Union Pacific Line was obscured for a bit this morning where the northbound side of the road re-emerges into tossed-coffee-cup range of the Burnett Transit Center light-rail stop (atop the topmost overpass in the shot above.) The semi that briefly plugged the hole looks to have scraped its way through the entire length of the tunnel before getting stuck at the northern exit. Transtar pinned the stopup to about 10:38 this morning; not much had changed as of 40 minutes ago (see below), but the road is now marked by professional traffic-watchers as cleared. The rapidly disappearing elevated segment of Elysian St. pointing north out of Downtown is the latest aging roadway structure to be crumbled apart, though it won’t be the last. But death is a natural part of the Houston roadway cycle! And a healthier, brawnier replacement viaduct is planned to take its place along roughly the same right-of-way — this one with broad shoulders and a sidewalk. TxDOT spokesman Danny Perez told Houston Public Media‘s Gail DeLaughter last month that work on the new structure, which connects Downtown to Near Northside by funneling drivers over Buffalo Bayou and I-10, should start before the demo of the mile-and-a-half-long original wraps up. A hunched excavator was spotted helping to bring the aging bridge down from above:

Comparison of stress in anaesthetic trainees between Hong Kong and Victoria, Australia. A postal survey was sent to anaesthetic trainees in Hong Kong and Victoria, Australia to compare work-related stress levels. Demographic data were collected. Anaesthetist-specific stressors, Maslach Burnout Inventory and Global Job Satisfaction scores were used for psychological testing. The response rates from Hong Kong and Melbourne were 64 of 133 (48.1%) and 108 of 196 (55.1%), respectively. Victorian respondents were older with greater family commitments, but more advanced in fulfilling training requirements. Hong Kong respondents, being faced with both the challenge of dual College requirements, exhibited consistently higher indices of stress (P < 0.001) and less job satisfaction (P < 0.001). Common occupational stressors related to dealing with critically ill patients and medicolegal concerns. Higher stress scores observed in Hong Kong trainees related to service provision and a perceived lack of resources. Despite the complex nature of stress, its antecedents and manifestations, an inverse relationship between emotional exhaustion and job satisfaction was evident in correlation analysis (P < 0.001). This survey suggests that stress was present in some trainees in both areas. Hong Kong trainees may benefit from local development to address mental wellbeing as being important to fulfil this highly competitive training program.

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B.A.P’s Helping Hands, a non-profit charity group led by fans, has raised over $26,000 since January 26, surpassing the original goal of $25,000. The group is teaming up with a non-profit organization called Pencils of Promise to build a school in Ghana for children in need. [Update] BABYZ blow us away!! Already making progress towards WASH ?? Donations now $26,413! https://t.co/aRqdFzhWGS @BAP_Daehyun pic.twitter.com/BuIo0L3cKE — BAP's Helping Hands (@BAPHelpingHands) November 24, 2016 The idea of building a school came from none other than Bang Yong Guk, the leader of B.A.P. Well-known for his charitable activities, Bang Yong Guk has led B.A.P’s efforts in donating books, funds and other necessary items to those in need. He stated that one of his goals was to build a school in Africa in B.A.P’s name. Fans from all over the world decided to make his dreams a reality by starting a donation fund with Pencils of Promise. Bang Yong Guk is aware of the project, and was even given the opportunity to name the school as the “B.A.P Town School.” Though the goal of $25,000 has been met, B.A.P’s Helping Hands will be taking donations till their deadline of December 26 to provide additional resources. Over the years, K-pop fandoms have become more involved in charitable causes to have a positive impact in the name of their favorite celebrities. From planting trees to building schools, K-pop is helping to make the world a better place. Congratulations to B.A.P’s Helping Hands and BABYz for accomplishing such a fantastic goal! Thanks to Marnie and Susanna for the tip!

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Knowledge-based potential defined for a rotamer library to design protein sequences. A knowledge-based potential for a rotamer library was developed to design protein sequences. Protein side-chain conformations are represented by 56 templates. Each of their fitness to a given structural site-environment is evaluated by a combined function of the three knowledge-based terms, i.e. two-body side-chain packing, one-body hydration and local conformation. The number of matches between the native sequence and the structural site-environment in the database and that of the virtually settled mismatches, counted in advance, were transformed into the energy scores. In the best-14 test (assessment for the reproduction ability of the native rotamer on its structural site within a quarter of 56 fitness rank positions), the structural stability analysis on mutants of human and T4 lysozymes and the inverse-folding search by a structure profile against the sequence database, this function performs better than the function deduced with the conventional normalization and our previously developed function. Targeting various structural motifs, de novo sequence design was conducted with the function. The sequences thus obtained exhibit reasonable molecular masses and hydrophobic/hydrophilic patterns similar to the native sequences of the target and act as if they were the homologs to the target proteins in BLASTP search. This significant improvement is discussed in terms of the reference state for normalization and the crucial role of short-range repulsion to prohibit residue bumps.

Hello, after a whole day puzzling without solution i ask for help. I have a Netgear NightHawk router R7000 and with their updated certificates for VPN service with OpenVPN I have my iPad pro (IOS) as well as my Samsung Galaxy phone (android) up and running perfectly as clients to my router, on which the VPN server is running (with mynetgear.com as the provider). To my surprise, though, it seems impossible to get my Windows 10 (64 bits) ASUS Zenbook laptop up to the same level. After installing the OpenVPN GUI (the newest as well as the "last stable") I get (visible in the log) the message "WARNING: No server certificate verification method has been enabled. See http://openvpn.net/howto.html#mitm for more info." Might it be something with the cooperation between the TAP-Windows adapter and my laptop, which has NO ethernet connection and only WiFi? In the adapter screen I notice that the (renamed) NETGEAR-VPN, the connection with the TAP-Windows adapter, needs the ethernet cable to be attached. (Needless to say that the field "Connected to the internet" stays blank....) Any suggestions? Anyone? Thanks for listening! Thanks for answering. But i do not onderstand why this would be only a Windows issue (the connection, with the same keys and certificates, is flawless in IOS and Android). But apart from that: how to build in that kind of security? I am afraid that I haven't got the skills to do that. Or is there a simple way to actieve this? I read it al right, that’s why i am afraid i juist haven’t the skills to do that. Apart from that, since my router acts as the VPN-server, i think i am not even allowed to mess with the router, plus i would not know where exactly to installe the server certificate. Maybe i should ask netgear about this all... But thanks a lot anyhow for thinking along! And might you have some otter suggestions, please let me know! My knowledge improves with every comment (and of course there is a lot room for improvement!) His/her name is Netgear and i will (I think that you"ll understand the concept of the impossibility for end users to change root programs / software of Netgear routers). If this leads to a solution i'll report about it at this place. TinCanTech, i try to be and stay polite... I think the point is perfectly clear. I can’t get my Windows 10 VPN connection to work, contrary to the IOS and Android connections, which work flawlessly. As an end user I can’t install a server certificate on my Netgear router, and I read the link in the OpenVPN.net/howto concerning the mitm and tried the extra line in my config (client1.ovpn), to no avail. That is also why I thougt it might be a TAP-Windows problem, which I mentioned in the title in this topic. The topic is not null and void, because i experience a problem which until now is not solved. I like to think that this forum is meant for these questions. I appreciate your responses. Your last remarks may express the difference between us regarding technical skills and knowhow, and that is the reason why i ask questions and you give answers. Sorry that you are apparently getting impatient. thanks again, and now extra because you really put some time in answering the newbie i am. Reading your links about howto request help, I am afraid that, in my router, I can't find the probable deeply hidden server certificate or related software /code. So I can't provide all the info you would need to help, and I won't ask you to put more time in this. I'll contact the Netgear people, because there is the chance that, while IOS and Android cause no problems, it is the Windows 10 connection that does not work. And of course, the server certificate is on their side. And by the way, it really does not work, it is not just the warning. No connection comes off the ground at all. As promised, because i started this topic, i shall post here the solution, if it comes. If you think it useful that i send to you only my client config .ovpn file and the client log lines, i will happily send them, also information about / screenshots of my router settings if that would help. Anyhow, thanks for your help, I really appreciate it.

Q: Website differs local from hosted - Different rendering of Jquery I've created a JQuery plugin which is creating an easy-to-use image gallery. The gallery looked just fine on my local server(I'm using Wampserver) and I uploaded it and it was rendered perfectly. Now a couple of weeks later they look like they should on the webserver, but not on my local. I've tried creating new documents, but they all render the same. Then I uploaded one of these documents and so it rendered perfectly on the webhost, but then again not on my local. Why's that? And how can I fic this problem? I couldn't find any information about this online and I'm sure of that there must be other people who have got this problem aswell. NOTE: AS FAR AS I KNOW THIS IS ONLY HAPPENING IN GOOGLE CHROME! BOTH MY LOCAL AND THE WEBHOST USES THE SAME VERSION OF JQUERY! Render Problem: In my gallery one of the transition effects is "slide". On my local they now slide more than what they should and they starting position is not where it should have been. If I slide from the right their position is a bit to the left of the starting position. But on the webserver they start on their normal starting position and they do not slide more than what they should. I've tried these "fixes": Clearing my cache(CTRL + F5) on Google Chrome Creating new documents Trying it on IE10 (If that's the one Win8 is using) - works Clearing all webinformation(except browsing history) on Google Chrome Uploading it to the web - works My Questions: Why doesn't it render it as it should on my local drive? And how can I fix this? A: I've had this problem before, and had to install a Chrome extension to properly trash the cache. I used Clear Cache, though many should work.

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Ralph Nader at Nader.org: The word “inequality” is much in vogue these days. We hear almost daily about the inequality of wealth, income and wages between the richest top 2 or 3 percent of people and the majority of the country’s wage earners. But not much attention is given and not many marches and other protests are addressing the huge inequalities between creditors and debtors. Of course the aforementioned inequalities, especially of wages and income, worsen the plight of individual debtors. One more distinction needs to be made – that between corporate debtors who receive many favored legal entitlements (even in bankruptcy) and individual debtors who are slammed and harassed by debt collectors. Start with the Federal Reserve’s low-interest policy of the last five years with no end in sight. Savers who used to get interest of 4 to 5 percent from their bank or money market now get, if they are lucky, ¼ of one percent on their savings. This Fed policy is supposed to stimulate the economy but doesn’t work very well if there is not enough consumer demand in a recession to attract new investment. Meanwhile, the hundreds of billions of dollars held by small, middle to low income savers are generating no interest to help pay their living expenses. The situation is bad and getting worse. These savers are being turned into “lockbox customers” in peril of having to actually pay the banks to hold their money. The Financial Times reports that “leading US banks have warned that they could start charging companies and consumers for deposits” if the Federal Reserve cuts interest rates further. Why don’t all those bellowing Congressional deregulators of health and safety standards ever object, except for the pure Ron Paul libertarians, to the overreaching Federal Reserve, the biggest market regulator of them all? Look at how the U.S. government has treated students borrowing for their education. When the government was not guaranteeing the gouging interest rates and fine-print traps of Sallie Mae and other corporate lenders that still have the iron collar around millions of college graduates, Uncle Sam was directly making money from students with interest rates around 6 percent. Other western nations offer tuition-free higher education as a great investment for their societies. Skyrocketing student loans now exceed credit card loans outstanding – $1.2 trillion in student loans compared to $1 trillion in credit card loans. With her proposed “Bank on Students Loan Fairness Act,” Senator Elizabeth Warren wants to reduce the student interest rate to the same rate paid by large banks borrowing from the Federal Reserve Bank, less than one percent. It is stunning how shortsighted this policy of gouging student borrowers is for the health of the economy. Their loan burden after graduation is such that they are less able to buy homes and cars in their twenties and thirties. In his fine new book Debtors’ Prison, Robert Kuttner recounts the history of debt, including the centuries when under Anglo-American law debtors were imprisoned or executed. He also describes how large corporate debtors today get bailed out or go through bankruptcy proceedings that save the company under a sweetheart rebirth process, complete with allowing executive compensations past and present. The individual debtors, however, are driven deeper into debt with fiendishly high interest rates (as high as 30% on unpaid credit card balances to over 400% on rolled over payday loans and rent-to-own rackets). Then there are the hundreds of different fees, penalties and costly fine-print impositions that ravage consumer borrowers. The profits from the credit industry were illustrated this week by MasterCard’s announcement. Its stock is up twentyfold to nearly $800 per share since it went public in 2006. Its profits are enormous, its dividends are surging, stock buybacks robust and there is no end in sight for its upward spiral. For a supposedly staid banking function, MasterCard acts like Apple. The sheer complexity of borrowing, paying, and getting some so-called relief or refinancing camouflages its own kind of costly exploitation. If the debtors object, particularly in a persistent manner, over such obvious greed, their credit scores – the new serfdom – can go down and put more burdens on their pocketbooks and future livelihood. A split U.S. Supreme Court endorsed the compulsory arbitration clause in these fine-print contracts that attaches more shackles. A report by Public Justice (“Wake Up!” can be seen here) found compulsory arbitration allowed predatory lenders to violate federal and state laws that protect consumers and blocked vulnerable elderly patients who had been abused in nursing homes from adequate access to the courts. Rays of help are coming from the enforcement of the Credit Card Accountability, Responsibility and Disclosure Act of 2009 by the young Consumer Financial Protection Bureau (CFPB). Over-limit fees and repricing actions are mostly eliminated and the dollar amount of late fees is down. Bait-and-switch to higher pricing once the borrower has signed on the dotted line has been diminished. In its latest report on the law (The CARD Act Report can be seen here), the CFPB recognizes other areas that “may warrant further scrutiny.” These include “add-on products” to credit card users, “fee harvester cards,” “deferred interest products,” and other problems stemming from the relentless ability of corporate lawyers to game and obfuscate the regulatory process and escape prohibitions with new avaricious coercions. Credit unions, with their ninety million members who are allegedly the owners, should be taking the aggressive lead in denouncing bad practices and thereby bringing even more consumer cooperators into their organizations. Less imitation of commercial banks and more dedication to cooperative service principles should be what members demand from this potentially large reform institution in our country.

Compact 1×4 wavelength demultiplexer based on directional coupling of periodic dielectric waveguides. A compact 1×4 wavelength demultiplexer is proposed based on the directional coupling of periodic dielectric waveguides for optical communication wavelengths. With appropriate optimization, the 1×4 wavelength demultiplexer can route 1130, 1310, 1490, and 1700 nm wavelengths to corresponding out ports with a transmittance of more than 95%. This provides a simple and compact demultiplexer that is expected to be applied to highly dense photonic integrated circuits.

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Sex change (disambiguation) Sex change is a natural or artificial process in which an individual's sex is changed. The term may refer specifically to: Sex reassignment therapy Sex reassignment surgery Sequential hermaphroditism, a phenomenon whereby some animals naturally change sex Sex change may also refer to: Sex Change, an album by Trans Am Sex Change or Change of Sex, a 1976 Spanish film A skateboarding trick involving a kickflip with a body varial

This invention relates to golf bags and in particular to golf bags having integral support stands. When a golf bag is carried by a golfer or a caddy it is desirable that it include a stand which will support it in a generally upright position where the golf clubs are readily visible for selection and can easily be removed and reinserted into the bag. This function has been accommodated in the past by providing legs that are movable between a retracted position where they rest against the side of the bag, and an extended position where they extend outwardly from the bag. Thus the legs do not interfere with carrying the bag when they are retracted, and yet act in cooperation with the base of the bag to support it when they are extended. Several methods have been used to move the legs between their retracted and extended positions. One method, which is used in a bag sold by Sun Mountain Sports of Missoula, Montana, connects the legs to the shoulder strap which is used to carry the bag. The legs are moved to their extended position by pulling on the shoulder strap and are moved back to their retracted position by a retraction spring when the bag is lifted off of the ground. Due to its complexity, this mechanism is expensive to manufacture and is easily damaged. In addition, this mechanism is awkward to use, particularly until the golfer becomes familiar with its operation. A simpler prior art device actuates the legs with a rod which extends below the bottom of the bag, the rod is connected to the legs such that the legs automatically extend when the bag is positioned with the rod resting on the ground and then is urged downwardly. This device is shown in the form of an attachable stand in Jones, U.S. Pat. No. D 283,339, However, with the device shown in Jones '339, the rod must extend below the bottom of the bag in order to make it serve its intended use, and thus it prevents the bag from being placed upright on the ground or in a golf car on its base. This is not a serious problem with the attachable stand disclosed in Jones '339, since the stand can easily be removed if the bag is to be carried in a golf car. However, this problem is a serious impediment to incorporating this type of stand actuation mechanism integrally into a golf bag. The subject invention permits incorporating the leg actuation system of Jones '339 integrally in a golf bag by relieving a portion of the base of the bag adjacent to the actuation rod, and making the rod terminate coplanar with the unrelieved portion of the base. The relieved portion is arranged such that when the bag is tilted to approximately the angle it will be at when the bag is supported on the legs, the end of the rod extends below the base and thus is exposed so that it can be moved when the bag is urged downwardly. Accordingly, the bag can be positioned vertically on the unrelieved portion of the base without interference with the actuation rod, and still have the actuation rod exposed so that it can be used to actuate the legs. In a preferred embodiment of the invention, the base of the bag is flat. The unrelieved portion is oriented normal to the center line of the bag and the relieved portion is angled relative to the unrelieved portion at approximately the same angle that the bag will be offset from the vertical when it is supported by the legs. Thus, the relieved portion also provides a platform which makes the bag stable when it is supported by the legs. Accordingly, it is a principal object of the present invention to provide a golf bag having integral legs which act in cooperation with the base of the bag to support it at an acute angle with respect to vertical. It is a further object of the present invention to provide such a golf bag in which the legs are moved to an operative position where they support the bag merely by urging the bag downwardly. It is a still further object of the present invention to provide such a golf bag in which the apparatus which is used to move the legs to their operative position does not prevent the golf bag from being supported vertically on its base. It is a further object of the present invention to provide such a golf bag having a base with a relieved portion which is angled relative to the unrelieved portion at approximately the same angle the bag will be offset from the vertical when it is supported by the legs. The foregoing and other objectives, features and advantages of the present invention will be more readily understood upon consideration of the following detailed description of the invention taken in

The Introduction of Marcus Cooper The Introduction of Marcus Cooper is the debut studio album by American recording artist and former Pretty Ricky vocalist Pleasure P; it was released on June 9, 2009, by Swagga Entertainment, Bluestar Entertainment and Atlantic Records. The title refers to the singer's birth name. The album peaked at number 10 on the US Billboard 200 and number 2 on Top R&B/Hip-Hop Albums charts, selling 19,000 copies in its first week. As of January 2011, the album has sold over 150,000 copies nationwide. The album was nominated for Best Contemporary R&B Album in the 52nd annual Grammy Awards, and the song "Under" was nominated for Best Male R&B Vocal Performance and Best R&B Song. Track listing Chart positions References Category:2009 debut albums Category:Pleasure P albums Category:Albums produced by Rico Love Category:Atlantic Records albums

Contents Summary Arnold makes a date with Ruth on Valentines' Night, then reads a letter from his French pen pal Cecile that she's coming over the same night (which is really Helga in disguise), and dates are set up at restaurants right across from each other. Arnold finds himself running back and forth between the two restaurants, until the real Cecile decides to surprise Arnold by flying over from France. By the end of the evening, both Helga-as-Cecile and Arnold have formed something of an attraction towards each other; Arnold is disheartened that he will never see 'Cecile' again, whilst being blissfully unaware that he has actually fallen in love with Helga by celebrating Valentine's Day. Before they leave, Arnold tells Helga, "We'll always have Chez Paris." This is a reference of the famous line in the 1942 film Casablanca, when Rick says to Ilsa before she leaves him: "We'll always have Paris." Arnold keeps one of Helga's shoes. A reference to the story of Cinderella. Jamie Mitchell got the directing credit, yet he's not credited as either a storyboard director or an animation director. Despite being a Valentine's Day special, the episode aired 4 days before the day itself. When Ruth smiles in the first scene, she doesn't have braces on her teeth. When Arnold is describing Ruth he says that she has 'soft BROWN eyes that sparkle', but Ruth is regularly show with BLUE eyes. PS 118's ZIP code is 96374-0171, which is listed as an Army Post Office code. Also, a number beginning with 96 usually means it's somewhere in the Pacific or Asia. Helga's hair ends up going from poofy to being worn down while she is vomiting in the bathroom, even though we never see her change it. When Helga is in the restaurant, the man sitting behind her is dressed in white. However, when they show Helga close-up, the same man is suddenly wearing black. There's no credit for Carla, the girl Gerald talked to who gave him the brush-off, not even another girl calling out to Ruth at the beginning. After Helga runs to the bathroom and before Arnold runs out to re-meet Ruth, he is holding a spoon full of pink food, the same as Helga but, he is eating what looks like brown steak, green peas and white and yellow potatoes.

[The effect of massage before venipuncture on the reaction of pre-school and school children]. The objective of this work is to describe the effect of massage done by parents on the reaction to venous puncture of preschooler and school age child hospitalized. Children's reactions were evaluated through the data of vital parameters, non-verbal communication and verbalization. The results obtained indicated that massage had significant effect in non-verbal reactions, especially those related to muscular relaxation. Effects on the reaction of vital parameters showed no difference between the two procedures, with and without massage, realized on the same child.

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While it would have been a bigger story if British critics had mercilessly slammed Hamilton, the blockbuster Broadway musical that has finally arrived at London's Victoria Palace Theatre, it's been showered with nothing but praise following its official opening last night. Critics have lined up to hail the show, a hip hop retelling of the rags to riches story of American founding father Alexander Hamilton, a masterpiece of musical theatre, echoing the feelings of audience members lucky enough to attend the show's previews earlier this month. Here are just some of the responses so far... Dominic Cavendish, The Telegraph: “The Greatest Show on Earth”, one paper declared the other day. C’mon! But seriously folks, there’s going to be more where that came from, because – lock up your doubters: I have to report that it really is as good as we’ve been told. Can you quibble that it’s a bit too cold, a bit too clever, and a bit too crammed with exposition? Perhaps, but it displays an artistic sophistication that makes most of its British counter-parts look like they’re mumbling into their shoes in comparison. Tony Peters, Radio Times:

// Boost.Bimap // // Copyright (c) 2006-2007 Matias Capeletto // // Distributed under the Boost Software License, Version 1.0. // (See accompanying file LICENSE_1_0.txt or copy at // http://www.boost.org/LICENSE_1_0.txt) /// \file container_adaptor/vector_adaptor.hpp /// \brief Container adaptor to easily build a std::vector signature compatible container. #ifndef BOOST_BIMAP_CONTAINER_ADAPTOR_VECTOR_ADAPTOR_HPP #define BOOST_BIMAP_CONTAINER_ADAPTOR_VECTOR_ADAPTOR_HPP #if defined(_MSC_VER) && (_MSC_VER>=1200) #pragma once #endif #include <boost/config.hpp> #include <boost/bimap/container_adaptor/sequence_container_adaptor.hpp> #include <boost/mpl/aux_/na.hpp> #include <boost/mpl/vector.hpp> namespace boost { namespace bimaps { namespace container_adaptor { /// \brief Container adaptor to easily build a std::vector signature compatible container. template < class Base, class Iterator, class ConstIterator, class ReverseIterator, class ConstReverseIterator, class IteratorToBaseConverter = ::boost::mpl::na, class IteratorFromBaseConverter = ::boost::mpl::na, class ReverseIteratorFromBaseConverter = ::boost::mpl::na, class ValueToBaseConverter = ::boost::mpl::na, class ValueFromBaseConverter = ::boost::mpl::na, class FunctorsFromDerivedClasses = mpl::vector<> > class vector_adaptor : public ::boost::bimaps::container_adaptor::sequence_container_adaptor < Base, Iterator, ConstIterator, ReverseIterator, ConstReverseIterator, IteratorToBaseConverter, IteratorFromBaseConverter, ReverseIteratorFromBaseConverter, ValueToBaseConverter, ValueFromBaseConverter, FunctorsFromDerivedClasses > { typedef ::boost::bimaps::container_adaptor::sequence_container_adaptor < Base, Iterator, ConstIterator, ReverseIterator, ConstReverseIterator, IteratorToBaseConverter, IteratorFromBaseConverter, ReverseIteratorFromBaseConverter, ValueToBaseConverter, ValueFromBaseConverter, FunctorsFromDerivedClasses > base_; // Access ----------------------------------------------------------------- public: vector_adaptor() {} explicit vector_adaptor(Base & c) : base_(c) {} protected: typedef vector_adaptor vector_adaptor_; // Interface -------------------------------------------------------------- public: BOOST_DEDUCED_TYPENAME base_::size_type capacity() const { return this->base().capacity(); } void reserve(BOOST_DEDUCED_TYPENAME base_::size_type m) { this->base().resize(m); } void resize(BOOST_DEDUCED_TYPENAME base_::size_type n, BOOST_DEDUCED_TYPENAME ::boost::call_traits< BOOST_DEDUCED_TYPENAME base_::value_type >::param_type x = BOOST_DEDUCED_TYPENAME base_::value_type()) { this->base().resize(n, this->template functor<BOOST_DEDUCED_TYPENAME base_::value_to_base>()(x) ); } BOOST_DEDUCED_TYPENAME base_::const_reference operator[](BOOST_DEDUCED_TYPENAME base_::size_type n) const { return this->template functor<BOOST_DEDUCED_TYPENAME base_::value_from_base>()( this->base().operator[](n) ); } BOOST_DEDUCED_TYPENAME base_::const_reference at(BOOST_DEDUCED_TYPENAME base_::size_type n) const { return this->template functor<BOOST_DEDUCED_TYPENAME base_::value_from_base>()( this->base().at(n) ); } BOOST_DEDUCED_TYPENAME base_::reference operator[](BOOST_DEDUCED_TYPENAME base_::size_type n) { return this->template functor<BOOST_DEDUCED_TYPENAME base_::value_from_base>()( this->base().operator[](n) ); } BOOST_DEDUCED_TYPENAME base_::reference at(BOOST_DEDUCED_TYPENAME base_::size_type n) { return this->template functor<BOOST_DEDUCED_TYPENAME base_::value_from_base>()( this->base().at(n) ); } }; } // namespace container_adaptor } // namespace bimaps } // namespace boost #endif // BOOST_BIMAP_CONTAINER_ADAPTOR_VECTOR_ADAPTOR_HPP

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Is the Gazette’s Homepage in Violation of Copyright? Luckily for the makers of SpaceBalls and the staff of the Gazette, US Copyright law provides for fair-use exemptions for parody as, the law reasons, no rights holder would give permission for others to mock their work. You can see our April Fools front page here, even after 4/1/2009. It is updated with our current content, however, so most of the year it will have real stories. This is not an April Fools joke. Hello, did you like this article? Write for The Gazette! Open staff meetings are every Monday at 7:30 p.m. in The Daily Gazette office on Parrish 4th; You can also email us at editors@daily.swarthmore.edu. 39 comments right, Chris would probably be more polite to your face and never say that anyone should be aggressively ignored. but as he's demonstrated time and again through his columns, he is an ardent supporter of the status quo, which perpetuates the aggressive ignoring of women, queers, and people of color. you can pick which you think is worse. Yeah, as someone who agrees with the political views of neither Marc nor Chris, Chris has always seemed more level-headed, incisive, insightful and open-minded. I don't think he'd ever say that anyone should be "aggressively ignored," as much as he might disagree with them. Marc, Chris was one of the few people on this thread calmly encouraging constructive dialogue. Aggressively ignore me if you want, or engage me on the merits, but I think constructive dialogue should be encouraged — isn't that a rule of discussion on this forum, DG editors? As a former editor of both the DG and Phoenix (a small group, I think — Jack Keefe, are you out there?) who has taken his fair share of potshots at both papers while working for the other one, I wanted to point out a couple of things — (1) A major source of the tension here is that you have two papers that overlap in some of their goals but also have, historically at least, done different things very well. The DG was online well before the Phoenix and has continued to excel in that arena. Moreover, being a daily, the DG can cover stories just plain quicker (even if the Phoenix is online, I wasn't about to check it daily for updates.) The Phoenix has been printing for over 125 years and offers a venue for a more diverse types of journalism just by virtue of the fact that it's in print. The two also overlap in a lot of their coverage, and whereas when I was a first year you could have said that the DG's coverage was by no means comprehensive of campus events or happenings, it's gotten there recently. At any rate, there are brilliant people working for both staffs, and there's going to be competition. That's how it was all four years when I was at Swarthmore, from both sides saying and doing nasty things. My senior year, there was some discussion of efforts to improve the relationship between the two staffs — nothing came to fruition, but I think an open recognition of the competitive nature of the two might help. If I could go back and change anything, and if I could suggest it now, it would be for the two staffs to chill out and be constructive — I wish I'd done more of it. (2) There's been some discussion of this in the thread, but you'd be amazed how repetitive things at Swarthmore are. I spent a lot of time when I was eic of The Phoenix rereading old issues, and, guess what? People in 1983 bitched about the food, wrote about an exhibition visiting Swat, had a dorm decoration contest, and wrote editorials about the exact same things as in 2009 — including how terrible The Phoenix was, how it should be destroyed, etc. The quality of the paper ebbed and flowed based on the composition of the staff and editorial boards. This is part of the nature of a college where students are only there for four years. (4) Just to put things in perspective, I left Swarthmore and went to law school, and I have to read this every week: http://www.gulawweekly.org/ . The students who work on it are really dedicated, but the quality isn't anything like that of the DG and Phoenix. (5) I don't always agree with the editorial decisions the DG staff makes (just as I'm sure they and the campus disagreed with many Phoenix editorial board decisions during my time), but I have to say that Miles and Lauren and the DG staff have done a great job improving the DG the past couple of years. Print media's in a lot of trouble, and getting this online system flowing is important in remaining relevant for ANY publication that's historically been in print, because that's where the future for media is. (6) Editors of both publications should be familiar with and use http://www.splc.org/ — great legal stuff. Journalists that often misquote or misinterpret quotes. And journalists who understand that as students, we are not necessarily skilled at delivering sensible-sounding "sound bites." I really appreciate it when the Phoenix writers take the time to confirm quotes that they are using. What journalist would ever do that? Journalism may not be about "tricking" people into saying things, but when you allow your subjects to retract what they themselves said, then you are just not being responsible. You're right that my column last semester didn't have specific solutions presented; my primary goal in writing that column was to show Swarthmore through the eyes of a guinea pig who could be considered conservative on a number of levels. It was fiction, and meant to give an impression and tell a story; my goal (whether successful or not) was not to grind specific axes, but to increase understanding of an alternative experience and perspective of life at Swarthmore. It was not intended to be a normal editorial column. Does that distinction make sense? (My column this semester is much more of a normal editorial, and I have attempted to include specific solutions where I can.) While registering discontent is fine, and admitting ignorance of a solution is okay, I have also found that sometimes the best way to resolve my discontent is to change myself (my attitude, my actions) and not the world. For instance, for a week I lived at a rural Chinese orphanage and ate beans and rice and plain white bread. Whenever I get sick of Sharples, I cast back on that week and realize what a blessed life I really do live. Let's be willing to strive for improvement, and yet always be grateful for the riches we have already. I think that the news content is fine in both the daily gazette and the phoenix and that the journalistic content is great(e.g. the phoenix's series for black history month). However the editorials at both papers (esp. the phoenix) suffer from a lack of creativity – with notable exceptions (fletcher's column this semester is amazing.) And Chris: you often criticize people for complaining about Swarthmore without offering solutions to the problem. Wasn't your hamster column pretty much your space to complain about aspects of Swattie culture you don't like? Those complaints weren't always accompanied by detailed solutions, many simply registered discontent. I don't think that's illegitimate, do you? I think the criticism of the Phoenix and Daily Gazette is a bit unwarranted. Look around for what passes for campus journalism at other similarly sized colleges. In many cases, they can barely (if even) manage to publish a paper on a regular weekly basis, let alone a paper and a daily news publication and a superb podcast such as War News Radio. Is every issue of the Phoenix and the Gazette scintelating? No. Is every article great? No. To be honest though, there is some superb stuff in both. For example, the coverage of the recent anti-abortion philosopher's talk was excellent in both the Phoenix and the Gazette — better than almost any mainstream media treatment of the topic. The reporting on financial issues and pending budget cuts has been solid, better than in many campus papers. And, Lauren Stokes has written some historical review articles that are simply excellent by any standard, such as her history of coming out week chalkings which in turn spurred her history of LGBT issues at Swarthmore. Great stuff. "(and I suspect the number of our college reporters who are breaking their teeth here just in preparation for journalism school is close to zero)" While very few go to Journalism School (of which many journalists are rather dismissive), quite a few Swatties go into journalism. We've got at *least* 5 Swatties at the New York Times right now, and there are more at a lot of alums at newspapers/radio stations/TV news stations all over the country. I'm glad for your reply, but I don't understand some of your comments. First, what are the problems you see with the style and format of major corporate newspapers? How should the format or style of Swarthmore's papers be different? You also say, "[The Phoenix] should function as an advocate for students and should give them its support." There are many institutions at this school that are supposed to function as advocates for students. Why should a newspaper, the goal of which should be to simply print the news as well as it can, take on a role as an student advocate? Also, in the same paragraph you complain both that The Phoenix writes world analysis/policy editorials and the same complaints about Swarthmore's campus and community. If you are suggesting, as seemed the case earlier, that Swarthmore's papers should focus on the community, won't it naturally follow that topics of continuous news will re-appear? You suggested that "The Phoenix ought to be brainstorming interesting, original, and important things to print…" — but what should they print? If you were running the phoenix this week, what articles would you publish that fits your criteria? I'm not disagreeing that there are weaknesses, flaws, and limitations in our papers, but I think this is more a reflection of the fact that we are a small school with little news that isn't already known by everyone, and without a journalism program (and I suspect the number of our college reporters who are breaking their teeth here just in preparation for journalism school is close to zero). it makes sense that our papers will be less than professional. I agree that there is room for improvement (especially, as you point out, on some specific themes that are over-worn, such as awkwardness at Swarthmore), and editorial guidance. Nevertheless, there has been improvement: The Phoenix misquotes people much less than it did four years ago. The Daily Gazette prints a greater number relevant stories than it did four years ago. Also, the sports section of the Phoenix is serves a valuable and fairly unique role in reporting college sports news on a campus generally apathetic to it. You're right that more work could be done to edit articles in both papers, and that editors could try to provide more of a structure and purpose to the papers. But they are also limited by the fact that their staff (and they themselves) are Swarthmore students, who submit things late, don't like covering events they aren't personally interested in, and have a dozen other things to do. J, I can't help but think my post was more than relevant to our discussion, especially because I was replying in particular to Chris Green's question above in comment 15. I'll quote it, since a reformulation would only bastardize the man's prose: "For people who have gripes about The Phoenix and the Daily Gazette, what are your specific complaints? What are your specific solutions?" Saykm, turning this comment page into a personal insult of the Phoenix seems far and away poor taste. The issue isn't whether the Phoenix provides good, literate journalism, which it does, but whether this April Fools Joke constitutes a parody, which it also most certainly does. That's definitely not private correspondance. It wasn't addressed to private-citizen-Miles, it was gazette-editor-in-chief-Miles, making it a "letter to the editor," and therefore public. Plus would it have been "trashy" to post it up as an explanation had the Gazette followed through with the demand and all of the sudden the site disappeared? The reason it seems trashy/insulting is because the letter is dumb. If the request was actually in response to a legit copyright infringement (which it certainly thought it was), no one would be complaining. copyright infringement? i feel so bad for the phoenix. it's bad enough dealing with the pressure of being the world's premier and most important news source without having to track down the legions of fanatic imitators trying to make a quick buck ripping off of the phoenix website's dynamic and edgy layout for their own two-bit news operations. The Phoenix: Taking Ourselves More Seriously Every Day since 1881 The Phoenix: Stealing the "Most Popular" Feature from the New York Times Website and Hypocritically Hating on Copyright Infringers since 1881 don't get me wrong: the gazette is a by no means a stellar news source (and its discussion board is a haven for the worst kind of nonsense) but the phoenix is a JOKE PAPER. that it publishes at all is a testament to the supreme self-importance of the phoenix staff, who seem to believe that the role of a college newspaper is to provide "news" in a format, style, and tone that tries in every way to ape mainstream, corporate news sources. (and get them into journalizm school fingers crossed) the phoenix should be a paper that documents the life of the school. not just the institutional life of the school, but the life of the people who comprise the school's community. it should function as an advocate for students and should give them its support. instead, the phoenix publishes the full names of swarthmore students who are arrested. instead, the phoenix thinks its job is to provide a space on its editorial sheet for every policy nerd in the school to spill more ink outlining their mindblowngly prescient "analyses" of world events. instead, the phoenix publishes column after column accessing the same tired tropes every everloving week ("the dating scene suxx bro! paces is stupid! sharples sucks! these are all of the things in the world that i know how to write about!"). instead, the phoenix prints the world "awkward" an average of 50 times an issue. do you know how many trees the phoenix is killing every week just to remind you that "life" at "swat" is "awkward"? the phoenix ought to be brainstorming interesting, original, and important things to print, not flipping out over the infringement of their precious copyright for an april fool’s joke. undirected anger, pointless criticism, ineffective complaints, petty frustrations. Chris, these aren't some 'disgraceful' traits of Swarthmore students. Its just a basic feature of human nature. I mean, what else is comedy, including parody, anyway? (ever seen seinfeld?)You seem to have fallen for the illusion that human feelings have a responsibility to be 'specific' and 'constructive'. Criticism is as much a valid expression as congratulation or appreciation ('i love this place') and needn't be qualified to meet your bureaucratic standards. oh yeah. and no matter how redic Anna's letter was, it feels like a cheap shot to post it in public. We're in a tiny community here, and it amplifies the hurt from every public insult. It would've been enough to just ignore the letter and leave the parody up. For people who have gripes about The Phoenix and the Daily Gazette, what are your specific complaints? What are your specific solutions? So many people complain about so much stuff at Swat without actually bothering to say what are the particular bad things and how to enact realistic change. A similar example is the ubiquitous complaining about Sharples food. Be specific and constructive; otherwise we are simply petty, ineffective, mean-spirited, and a disgrace to ourselves and our college. Disregarding the relative and absolute journalistic merits of the two publications, it's still sad that y'all don't still have Nelson Pavlosky around to yell at the Phoenix until they admit the error of their ways. If Swatties could once educate large corporations on the fine points of fair use (along with a small brigade of lawyers), why can't we impart some clue into our own campus newspaper? While I'm not a huge fan of either paper, I have consistently found the Daily Gazette's reporting to be lacking of any modicum of responsibility or professionalism. At least reporters from the Phoenix actually email quotes to interviwees before printing them. By the way, it's Students FOR A DEMOCRATIC SOCIETY. Perhaps Swatties should take themselves less seriously. Obviously the layout is a joke, making fun of the alleged rivalry between the Gazette and the Phoenix, I highly doubt Miles meant it as a slight.

440 So.2d 1374 (1983) CITY COURT OF BREAUX BRIDGE, Plaintiff-Appellant, v. TOWN OF BREAUX BRIDGE, Defendant-Appellee. No. 83-326. Court of Appeal of Louisiana, Third Circuit. November 9, 1983. Writ Denied January 27, 1984. Durrett, Hardin, Hunter, Dameron & Fritchie, Ben W. Lightfoot, Baton Rouge, W. Glenn Soileau, Breaux Bridge, for plaintiff-appellant. McHugh & Guidry, Edmond Guidry, St. Martinville, for defendant-appellee. Before FORET, STOKER and YELVERTON, JJ. STOKER, Judge. By this suit the City Court of Breaux Bridge seeks to compel the Town of Breaux Bridge to pay certain expenses of the court including postage, printing of forms, telephone expenses, and the upkeep of the library. *1375 The case arose through the filing of an application for writ of mandamus by the judge of the City Court addressed to the district court for the district in which the City Court is located. The district court ruled adversely to the city court and dismissed the application. The City Court appealed the dismissal. We reverse. The Town takes the position, and the trial court so found, that its obligation to the City Court is limited by LSA-R.S. 13:2488.76 which provides: "The expenses of operation and maintenance of the courtroom and offices shall be paid by the town of Breaux Bridge." The trial court held that the expenses referred to in LSA-R.S. 13:2488.76 are confined to those necessary for the physical operation and maintenance of the City Court, not its operation as a court. Such expenses, according to the trial court, would encompass utilities, cleaning, painting, repairs, and upkeep. The trial court also found, as contended by the Town, that the expenses sought by the City Court in this suit were provided for in LSA-R.S. 13:2488.78 which states: "A. In all criminal matters, including traffic violation cases, the city judge may assess, in addition to the fine or other penalty imposed, costs of court in an amount not to exceed eight dollars. "B. The proceeds derived from these costs shall be deposited in a special account which shall be subject to audit and shall be used for the operational expenses of the court, including but without limitation, the use from time to time for the costs of operating the court and for the employment by the judge of secretaries, accountants, stenographers, filing clerks, bookkeepers, reporters and other court employees, and for the purchase of stationery, books, office supplies and such other equipment, all as may be useful or necessary for the proper conduct of the court's judicial business, and all as may be approved by the court. In addition, the proceeds derived from such costs in excess of that necessary for the proper conduct of the court's business may, with approval of the judge, be paid in equal proportions into the treasury of the town of Breaux Bridge and into the treasury of the parish of St. Martin." The trial court noted that its ruling put the City Court in an awkward position as it now had to find more criminal defendants guilty in order to pay its expenses. The trial court suggested that relief be sought from the legislature. On appeal the City Court contends (1) that the Town's obligations to pay City Court expenses are not limited to LSA-R.S. 13:2488.76 but rest upon principles flowing from constitutional separation of powers, and (2) in the alternative, that the expenses provided in LSA-R.S. 13:2488.76 are not limited as found by the trial court. We do not found our decision on the statutory authority considered and interpreted by the trial court. Rather we think the power of the City Court to require that all its necessary expenses of operating the court be provided by the legislative body of the Town lies in the inherent power of the City Court. A separate panel of this Court of Appeal has considered this same question relative to a district court of this state and the police jury of the parish which the district court serves. In W.T. McCain, Judge of the Thirty-Fifth Judicial District Court v. Grant Parish Police Jury, et al, 440 So.2d 1369 (La.App. 3rd Cir.1983), a panel of Judges Domengeaux, Guidry and Cutrer, in an opinion authored by Judge Cutrer, has held that "we ... find that courts have the inherent power to compel the guardians of the public fisc ... to budget adequate funds for the operations of the court to insure that the proper independence among our three co-equal branches of government be maintained." The members of this panel have considered, as a general proposition, the inherent *1376 power of courts of law to compel a branch of the same political body as the court serves which is responsible for funding the operations of that political body, to fund the judicial functions and operation of the courts. We arrive at the same conclusions which are expressed in the McCain case and adopt those conclusions as our own for purposes of this case. We do not regard the statutes referred to above as constituting a limitation or negation of the inherent power possessed by the City Court of Breaux Bridge. The provisions of those statutes simply lay down certain specific directions to the Town and authorize the assessment of court costs and the use of funds derived from such assessment. The remainder of this opinion will be addressed to the remaining issue raised by the Town. That issue is the contention by the Town that, regardless of inherent power, the City Court of Breaux Bridge in particular is not a court which enjoys such power. The Breaux Bridge City Court is not one which is expressly established by the Louisiana Constitution. The City Court was established by the legislature pursuant to its authority in the Louisiana Constitution of 1921, Art. VII, sec. 51. The City Court was retained in Art. V, sec. 15 of the 1974 Constitution. Under the new Constitution, the legislature may no longer establish city courts, but has the authority to abolish those in existence. The Town argues that because the City Court is "statutory" it does not have the inherent power accorded "constitutional" courts, i.e., those expressly provided for in the Constitution. We reject this argument. Art. VII, sec. 1 of the Constitution provides: "The judicial power is vested in a supreme court, courts of appeal, district courts, and other courts authorized by this Article." The Breaux Bridge City Court is one authorized by Article VII of the Constitution; therefore, it is vested with the same inherent power as courts directly and specifically established by the Constitution. We note that at least one other state has taken this same position. In City of North Las Vegas v. Daines, 92 Nev. 292, 550 P.2d 399 (1976), the Nevada Supreme Court stated: "It is, of course, true that our Constitution does not itself establish municipal courts. (Citations and footnotes omitted.) It authorizes the Legislature to do so. However, once municipal courts are established, they exist as a coequal branch of local government within the judicial department of this state, * * *, and a part of the constitutional judicial system of this state, * * *." This position was reaffirmed by the Nevada Supreme Court in Azbarea v. City of North Las Vegas, 95 Nev. 109, 590 P.2d 161 (1979). We find, as did the Nevada court, that city courts are a part of the constitutional judicial system and, as long as they exist, are vested with the same inherent power as courts specifically mentioned in the Constitution. The evidence regarding expenses of the City Court was admitted without objection. No claim is made that the expenses sought are not reasonable and necessary to the operation of the court, and it appears to us that the requested expenses are for the very basic services and supplies necessary to the operation of the court. Demand for payment of these expenses does not appear to be an abuse of the inherent power of the City Court. For the above reasons, the judgment of the trial court is reversed. IT IS NOW ORDERED, ADJUDGED AND DECREED that a writ of mandamus issue herein ordering the Town of Breaux Bridge, defendant, through its duly elected representatives constituting the governing authority of the Town of Breaux Bridge, to pay all reasonable unpaid expenses for the operation of the City Court of Breaux *1377 Bridge to include, although not by way of limitation, expenses for postage and mailing, printing of necessary forms, telephone costs, and upkeep and maintenance of the law library of the City Court of Breaux Bridge and other reasonable operating expenses germane to the operation of that court. IT IS FURTHER ORDERED, ADJUDGED AND DECREED that the effect of this order shall extend only to any net balance of expenses owed by the City Court of Breaux Bridge over and above any payments thereon which may have been paid by the City Court of Breaux Bridge itself. All costs of court in this matter including the costs of this appeal are to be paid by the Town of Breaux Bridge. REVERSED AND RENDERED; WRIT OF MANDAMUS ORDERED.

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1. Field of the Invention This invention relates generally to rolling mills, and is concerned in particular with an improvement to the finishing section of a single strand bar mill. 2. Description of the Prior Art The roll stands of conventional bar mills are usually aligned along a single pass line. Where slitting is being practiced, the roll stands of the roughing and intermediate mill sections are again aligned along a common pass line, with two sets of roll stands in the finishing section being aligned along parallel pass lines arranged to receive the slit sections. In both cases, the arrangement of the roll stands in the finishing section imposes severe limitations on the range of products the mill is able to produce.

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The candidate is an MD/PhD trained ophthalmologist and ophthalmic plastic surgeon with the career goal of studying stem cells and their application to cure ocular and orbital disease. The career development plan will be jointly mentored by Drs. Ihor Lemishcka and Mario Wolosin. Their laboratories focus of human embryonic and induced pluripotent stem cell biology and corneal limbal stem cells, respectively. The candidate's long-term aim is to understand what controls stem cell differentiation into distinct ocular and orbital tissues, with the goal of creating autologous tissue for transplantation. Over 6 million people worldwide are blind from damage to the ocular surface. Burns, trauma, infection, genetic diseases, and chronic inflammation result in limbal epithelial stem cell deficiency (LSCD) and persistent vision loss from corneal scarring, vascularization and conjunctivalization. Replacement of limbal stem cells is often required to restore vision. In unilateral LSCD cases, the fellow eye is an obvious autologous cell source; in patients with bilateral LSCD (in cases of bilateral blindness) limbal allografts are the only option. For reasons not fully understood, allografts often fail within a few years in these patients despite immunosuppression. Thus, autologous cells are necessary to restore lasting vision. We propose to develop a protocol to generate autologous induced pluripotent stem cell (iPSC)-derived cells capable of acquiring the corneal phenotype in vivo. Our aims are to (1) generate iPSC from cells whose embryological origin is closer to the limbal- corneal lineage, which includes human eyelid skin, and conjunctival and buccal epithelia, (2) differentiate iPSC via progenitor cells using an ectodermal differentiation protocol in combination with soluble factors and limbal niche culture, and (3) assess the ability of the generated cells for ocular surface regeneration using an established human tissue into rabbit model. The funding for this proposal will facilitate the discovery of new stem cell therapies for ocular surface disease and make regenerative medicine a reality for those burdened with blindness.

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Characterizations of heterotopic neurons in the spinal cord of amyotrophic lateral sclerosis patients. This report concerns a comparative immunocytochemical, ultrastructural and morphometric investigation on heterotopic neurons in the white matter of the spinal cords of 19 patients with amyotrophic lateral sclerosis (ALS) and 18 age-matched neurologically normal individuals. The study revealed that the heterotopic neurons were scattered in the white matter, often adjacent to gray matter, that they immunoreacted with the antibody to synaptophysin, and that there were synaptic apparatuses on the surface of their somata and their neuronal processes. Bunina bodies and ubiquitin-positive inclusions such as Lewy body-like inclusions and skein-like inclusions, characteristic of anterior horn neurons of ALS, were present in the cytoplasm of the patients' heterotopic neurons in the anterior or lateral column of the white matter. These findings suggest that heterotopic neurons in the anterior or lateral column have the characteristics of alpha motor neurons. The average number of heterotopic neurons observed in ALS patients was generally less than in normal subjects. This reduction was correlated with the severity of neuronal loss. The heterotopic neurons in ALS were less susceptible to the degenerative process as compared with spinal cord anterior horn cells. We assume that in this disease the heterotopic neurons may be degenerated and their number diminished after or concomitantly with the depletion of anterior horn neurons.

We have determined that wear particles retrieved from human joints stimulate macrophages and synovial cells to release enzymes which have the potential to degrade cartilage and other tissues of the joint. Further experiments will be directed towards identifying the factors which provoke this response of the cells to the particles. In addition,the enzymes which the cells secrete after particle treatment and which are able to degrade cartilage will be better characterized.

Genetics May Influence Kidney Failure In Blacks In a case of natural selection with a twist, genetic variants that confer protection against disease in Africa seem to place African Americans half a world away at elevated risk of kidney failure, a new study finds. U.S. blacks are three times as likely as whites to develop chronic kidney disease. “We think this explains perhaps most of it,” says study coauthor Martin Pollak, a nephrologist and geneticist at Harvard Medical School and Beth Israel Deaconess Medical Center in Boston. The gene in question is called APOL-1, short for apolipoprotein L-1. Among people of African descent, APOL-1 can appear in two variant forms that protect against African sleeping sickness but also increase susceptibility to kidney failure, Pollak and his colleagues report online July 15 in Science. Each parent provides one gene copy to offspring. The team analyzed blood from more than 1,400 African Americans and found that more than 30 percent carried at least one variant copy of APOL-1. About 10 percent harbored two variant copies. Text continues after gallery … Roughly half of the people in the study had kidney disease and half didn’t. The researchers found that those carrying two copies of an APOL-1 gene variant were 10.5 times as likely to have kidney disease marked by loss of blood filtration capacity compared with people who carried the normal APOL-1 gene. The double-copy carriers were 7.3 times more likely to have severe kidney disease marked by high blood pressure that necessitated dialysis or a transplant. People harboring two variant copies were also more prone to such end-stage disease compared with people having one variant copy. While harboring two copies of a variant seemed to increase risk, having one variant and one normal copy did not.

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Regardless of whether or not you want to see kids dying or not, making it so that you literally can't see what is happening at any point is NOT an improvement. I thought the film was great, though I never read the book, watched a trailer beforehand or read anything about it before seeing it. I just watched it randomly as we missed the showing of what we went to watch. So from my point of view I did not feel it was doing what you are saying, but the again I was watching it as a "hunger games noob", no expectations. Edit - At no point did I feel I was missing anything due to camera work. Regardless of whether or not you want to see kids dying or not, making it so that you literally can't see what is happening at any point is NOT an improvement. I thought the film was great, though I never read the book, watched a trailer beforehand or read anything about it before seeing it. I just watched it randomly as we missed the showing of what we went to watch. So from my point of view I did not feel it was doing what you are saying, but the again I was watching it as a "hunger games noob", no expectations. Edit - At no point did I feel I was missing anything due to camera work. Well, they specifically made the camerwork bad so that you couldn't see what was happening, so you definitely were. Regardless, I really hate any movie that has bad camera control, and The Hunger Games had some of the worst. Yeah, I don't agree with this list at ALL. There's a few that belong on there, but for the most part, pretty much all these films are forgettable. My top ten are as follows, mind you these are my favorite movies of the year, not necessarily the best. 1. Skyfall: I'm not versed in James Bond films, I've seen a few, but for the most part my knowledge of them is small. Skyfall made me love James Bond -- it was fun, smart, cool, exciting, funny, and the most pleasant surprise -- it was really well made. Most of the time, with an action film franchise like Bond, the filmmaking is lackluster and doesn't do much to elevate the themes or conflict of the film, but Mendes actually crafted this film with careful consideration, and it shows. It was a complete departure from classic Bond, but at the same time, served as the perfect homage to the series. 2. Django Unchained: I saw Django last night so I haven't had enough of time to really mull this film over, but it doesn't matter, because it's already one of the best of the year. As Moviebob said, this is probably most humanistic film Tarantino has ever made. It showcases his classic tropes with over-the-top characters, witty dialogue, and bloody - and I mean BLOODY - shootouts, but at the same time, Tarantino actually had more to say with this film than he has in a long time. 3. Cloud Atlas: This film was a behemoth. I was incredibly surprised at just how it turned out, considering the immense ambition behind it. The actors were all great playing their multiple parts and every separate story was interesting and kept me intrigued. Watching how everything unfolded was one of my favorite experiences at the movies this year. 4. Looper: This movie was perfect proof that action flicks don't have to be mindless dribble with cookie-cutter characters. The characterization was excellent and provided a satisfying emotional climax. 5. The Master: Huhhhh boy, where to start? Either you like Paul Thomas Anderson, or you don't. I love PTA. I love how he never provides interpretations or tells his audience what something means -- he wants THEM to figure it out and that's rare in film now-a-days. The Master was as fascinating as it was challenging. It wasn't a film made for entertainment -- it was a complex work of art. The only reason it's not higher on my list is beCAUSE it was so challenging and satisfied a strictly intellectual itch. 6. Moonrise Kingdom: This was actually my introduction to Wes Anderson, and I am so glad it was. I've now seen a number of his films and Moonrise Kingdom is still my favorite. Anderson has always been great at combining whimsy with very real, almost tragic circumstances and Moonrise is no exception. It's been a long time since I've been able to forget about how the world really works and get lost in a fantasy, but this film gave me that escape. 7. Cabin in the Woods: This is one I can admit to liking for no other reason than it being so much god damn fun. A horror film that slashes genre tropes to gory, bloody pieces while at the same serving as a commentary on the genre itself? Awesome. 8. Safety Not Guaranteed: The indie gem of the year. Well-structured with some very relatable characters and subtle comedy made me enjoy it immensely. 9. Beasts of the Southern Wild: One of the most powerful and poignant films of 2012. Beasts told a wonderful story of loss and understanding from the point of view of an innocent girl named Hushpuppy. Her connection with nature and the poverty-stricken lifestyle of her people is one we should all take something away from, and in a world that's becoming more and more digitized, this is a film to be remembered. 10. Lincoln: This film is last on my list only because I would've liked to see more out of it. Lincoln is great insight into the life of a truly incredible man, but it only focuses on a very small portion of his life as well as a LOT of politics that went on during the abolition of slavery. Personally, I take more interest in the man himself than the politics and ALL of Lincoln's life is a fascinating story, one I would've liked to see in its entirety. Nonetheless, Lincoln is a great film with some great performances and anyone who is interested in American history should love it. [Kira Must Die]:Alright, regarding SK3, do I have to have seen the other two films in order to "get" this one? Well, it's a direct sequel to the first one, so I'd recommend seeing that, as it's what's being "battle[d] for redemption" in the third one. The second one is a prequel to the first film, and is in no way, shape, or form worth seeing. Not even for the story. Not necessary. I do wonder if anyone is quite so annoyed by half assed one liners as I am. Because if I'm not the only one, why is Avengers so well received? Stringing together clichéd lines with cg effects and cheap laughs should not make a movie.But I might be on my own with this one. I'm avoiding pitch perfect, as I know what I like and what I don't, and that film definitely falls into the latter category, intrigued by cloud atlas though. Got to see Les Miserables with the Missus. They muffed a few things but the power of the story and music are incredible. No one should miss this movie. The wife didn't stop sobbing all day just thinking about it. I can't say I agree with someone putting Hunger Games ahead of The Hobbit. I'm no film connoisseur and even I know the main premise of it is ripped off from Battle Royal. Not saying it was terrible or anything its nice to see a teen film series that shows a young female lead thats not 100% dependent on others like a certain vampire series.... and the action scenes we're surprisingly brutal for the rating of the film. But the main premise of the film is clearly stolen from royal. Plus the hobbit was fantastic, at least as good as fellowship, I found the dwarf architectural design and traditional clothing a little jarring at first as its so alien, especially in the new frame rate, but once I got used to it, it was another emersive area of middle earth for me to settle into. One thing I was particularly happy with was to see that it still resonates with me when LOTR came out I was a young teen so was nice that I could still enjoy it without the nostalgia factor The best film of this year that I saw? Looper. Brilliant, interesting storyline with a sci-fi world that managed to appear futuristic without feeling ridiculous (Total Recall, anyone?). Film that I enjoyed the most? The Avengers. I was always a mark for this movie, seeing as I'm mainly a comic book nerd, but it managed to be one of the few films that I've been excited for that not only met my expectations, but actually went above them to provide a truly memorable experience. Didn't see any really bad movies this year. Prometheus was only OK, I guess. My favorite movie of this year was Snow White and the Huntsman. Notice how I didn't say 'best'. I loved it, though, mostly because of its very proud display of all that it could have been. Each new locale that the movie visited felt like it wanted to be a movie in and of itself, and just thinking about all of that potential has given me more joy than any other film in 2012. How is it that The Dark Knight Rises has 'stronger characters' than The Avengers? Come to think of it, how is it that The Dark Knight Rises has stronger characters than a shampoo commercial? Is that assessment based entirely around Michael Caine's performance and no other factors? I could maybe see it if that's the case. Here are my picks for 2012. Best Picture: Django UnchainedHonorable Mention: ParaNorman, Snow White and the Huntsman, The Hobbit: An Unexpected Journey, The Avengers, Ted Overall a pretty good year for movies, definitely. I had more good times than bad times, and The Dark Knight Rises being the nadir of the whole thing means that it could have been so much worse. Rises was pretty godawful, but it wasn't a huge disaster. Good thing we can disagree. I disagree with most of your Top picks. I'm not saying Hunger Games is good or bad but what makes it any more special than Twilight? And I thought you were joking about the first couple of picks but oh well.

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This month Montana will legalize the harvest of roadkilled deer, elk, antelope, or moose struck in car accidents. The buzzards aren’t happy, but people should be. Currently about two-thirds of states prohibit harvesting roadkill. Allowing people to harvest what they accidentally kill could help salvage millions of pounds of otherwise wasted meat. It provides a leaner alternative to factory-raised meat. It is also highly ethical since the wild animals were killed accidentally and never subjected to potentially brutal factory farming practices like extreme confinement, debeaking without anesthesia and painful transportation in jam-packet trucks. [pullquote] It will help to further reinforce the reality that meat does not come shrink-wrapped in grocery stores, but from a live animal that gave up its life. Allowing the meat to be consumed, rather than rot on roadways, also demonstrates a greater respect for animal life. Even PETA agrees that road kill is superior to factory meat, stating on its website: “Eating roadkill is healthier for the consumer than meat laden with antibiotics … It is also more humane in that animals killed on the road … did not suffer the trauma and misery” of factory farming. Who knew that road kill could unify hunters and PETA! According to the National Highway Safety administration there are about 1 million deer-related car accidents each year. Other estimates place it at closer to 1.5 million. An average deer will yield about 50 pounds of meat. That is potentially 50 million pounds of venison that could be eaten. An estimated 18 or so states permit some form of roadkill harvest with laws varying widely, from mere reporting requirements to more cumbersome permit application procedures. This means most of the nation including states like Texas, Washington, Tennessee and California generally prohibit salvaging roadkill. In terms of health concerns, according to Food Safety News, roadkill is similar to eating animals killed by hunting. So long as the meat is fresh, and cooked properly to kill bacteria, there are no additional risks. Wild meat has less fat and is generally healthier than hormone injected factory farm raised meat. The roadkill meat can also go to food banks to help feed the hungry. In 2010, for example, hunters provided nearly 2.8 million pounds of game meat to shelters, food banks and church kitchens, providing 11 million meals to the less fortunate. Some states already have programs that utilize roadkill to feed the hungry. All laws are subject to abuse and fraud. Givens the extensive damage to cars, risk to life, and increased insurance premiums that accompany collisions, it is unlikely that people will intentionally ram into animals just to harvest the meat. Reporting requirements, investigations where necessary, will further reduce the instances of potential abuse by motorist poachers. Bottom line: Every state should allow roadkill harvest. The meat is healthier than the antibiotics, hormones, growth stimulants and fat in our chicken nuggets and burgers. The harvest is highly ethical since the animals never suffered in captivity. Waste will be reduced. Animal life will be accorded the respect it deserves. It will compel people to appreciate that the animals give their lives to sustain us and don't come shrink-wrapped from grocery stores.

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Luke and Q Luke and Q are an African-American R&B duo from New Orleans, United States best known for their 2006 hit single, "My Turn". The members include Luke Boyd (now known as Luke James) and Quinten Spears. The duo met in eighth-grade when Luke overheard Q singing Ginuwine's, "Same Ol' G". As Luke recalls, "I was, like, You can sing, dog, but you may want to flip it," who then belted his version. They joined another childhood friend and formed a trio called Upskale, but the group disbanded almost as quickly as it began. The duo then began performing at local events, eventually singing background for R&B superstar, Tyrese. Through Tyrese, they met production duo Damon Thomas and Harvey Mason Jr. of The Underdogs for Underdog Entertainment, and were introduced to Clive Davis who subsequently signed them to the J Records imprint in 2004. They released their debut song called, "My Turn". Spears has been inactive in the mainstream music business since. Discography Singles 2006: "My Turn" References External links Luke and Q on Myspace Category:American musical duos Category:American contemporary R&B musical groups Category:Contemporary R&B duos

468 F.2d 1000 72-2 USTC P 9688 Jacob OPPEWAL et al., Petitioners-Appellants,v.COMMISSIONER OF INTERNAL REVENUE, Respondent-Appellee. No. 72-1126. United States Court of Appeals,First Circuit. Heard Sept. 6, 1972.Decided Oct. 3, 1972. Jacob Oppewal, pro se. Harold C. Wilkenfeld, Atty., Tax Div., Dept. of Justice, with whom Scott P. Crampton, Asst. Atty. Gen., Meyer Rothwacks, Gary R. Allen, and Joseph M. McManus, Attys., Tax Division, Department of Justice, were on brief, for appellee. Before COFFIN, Chief Judge, McENTEE, Circuit Judge, and HAMLEY*, Senior Circuit Judge. HAMLEY, Circuit Judge. 1 In their federal income tax return for 1968, Jacob and Leona G. Oppewal, of Whitinsville, Massachusetts, deducted as a charitable contribution, the sum of nine hundred dollars they had paid that year to the Whitinsville Society for Christian Instruction (Society). The Commissioner of Internal Revenue disallowed this to the extent of six hundred and forty dollars on the ground that this sum, representing the cost to the Society of educating taxpayers' two children that year, constituted a non-deductible personal expense under section 262 of the Internal Revenue Code of 1954 (Code), 26 U.S.C. Sec. 262. 2 On this ground the Commissioner declared a 1968 tax deficiency in the amount of $153.22. Taxpayers petitioned the Tax Court to redetermine the asserted deficiency. The Tax Court adhered to the Commissioner's determination, its findings of fact and opinion being reported at 30 T.C.M. 1177. Taxpayers appeal to this Court. 3 The controlling facts are not in dispute. The Society was organized in 1924 ". . . for the purpose of advancing the cause of education . . ." It was, during the taxable year 1968, an organization exempt from tax under section 501 of the Code, 26 U.S.C. Sec. 501. The membership of the Society consisted of three hundred and twenty contributing individuals and families, some ninety of which had children attending the school. The Society solicited and received gifts from members, non-members, churches and other organizations. 4 Members who were parents with children in the Society's school were solicited in the same manner as were non-parents. Money received was not put into donor or students accounts but was put into a general operating fund. No child obtained the opportunity to attend the Society's school by reason of any contribution made, nor was any child barred for lack of a contribution. Approximately forty percent of the total Society receipts was collected from the contributions of parents who had children in the school, though a record or tally was not kept. Funds were solicited on the basis of what a person could afford to give rather than on a per capita basis. 5 Funds normally came to the Society on a regular established basis rather than as a result of a drive or newly-initiated program. Solicitations were made from time to time on a personal basis. The academic training received at the Society's school was no better or worse than that available at the public schools in taxpayers' community, consisting of about ten thousand people. 6 During the 1967 to 1968 school year, one hundred and ninety-three children attended the school operated by the Society. The cost of operating the school amounted to about three hundred and thirty-eight dollars per student. During the taxable year 1968, two of the taxpayers' children attended this school for a period of thirty-nine weeks. 7 Section 170 of the Code, 26 U.S.C. Sec. 170, allows as a deduction any charitable contribution payment of which is made during the taxable year. Section 170(c) defines a "charitable contribution" as including a "contribution or gift to or for the use of . . . [a] corporation . . . or . . . fund . . . organized and operated exclusively for religious . . . or educational purposes." 8 The Tax Court, relying upon its decision in Harold DeJong, 36 T.C. 896 (1961), affirmed sub nom. DeJong v. C. I.R., 309 F.2d 373 (9th Cir. 1962), held that to the extent of six hundred and forty dollars, the taxpayers' payment to the Society was not a "contribution or gift" to the Society, because it proceeded primarily from the incentive of anticipated benefit to the taxpayers "beyond the satisfaction which flows from the performance of a generous act . . ." Said the Tax Court: 9 ". . . we are persuaded that the contributions made by petitioner to the Society in 1968 were substantially induced by the benefits anticipated by him from the enrollment of his two children in the Society's school and were, to a substantial extent, in the nature of tuition."1 30 T.C.M. at 1178. 10 In their brief in this court, taxpayers apparently accepted the subjective test applied by the Tax Court involving taxpayers' motive and intent in making the payment to the Society. However, they argued that, under the facts, it was not reasonable to conclude that the payment was induced by the educational benefits taxpayers expected to derive for their children. In oral argument taxpayers urged that objective rather than subjective standards should be applied. The objective test taxpayers would apply is whether there was certainty as to the beneficiaries of their payment. S. E. Thomason v. C.I.R., 2 T.C. 441 (1943). They assert that this test was not met because no part of the payment was earmarked for the education of taxpayers' children. 11 We have heretofore expressed our dissatisfaction with such subjective tests as the taxpayer's motives in making a purported charitable contribution. In Crosby Valve & Gage Company v. C. I.R., 380 F.2d 146 (1st Cir.), cert. denied, 389 U.S. 976, 88 S.Ct. 477, 19 L. Ed.2d 468 (1967), we said: 12 "While agreeing with the holding of the Tax Court, we think it necessary to register our disagreement with the [Tax Court] majority's emphasis upon a purely charitable motive as a prerequisite for a deductible charitable contribution. Were the deductibility of a contribution under section 170(c) of the Internal Revenue Code of 1954 to depend on 'detached and disinterested generosity', an important area of tax law would become a mare's nest of uncertainty woven of judicial value judgments irrelevant to eleemosynary reality." 380 F.2d at 146. 13 But while we believe it important to solve this tax problem through the application of an objective test, we do not regard taxpayers' proposed objective inquiry-was the payment earmarked for the education of taxpayers' children-appropriate. Classification of a payment as a gift, or as payment of family expenses, should depend upon something more fundamental than the particular bookkeeping methods used by the educational institution. 14 The more fundamental objective test is-however the payment was designated, and whatever motives the taxpayers had in making it, was it, to any substantial extent, offset by the cost of services rendered to taxpayers in the nature of tuition? If so, the payment, to the extent of the offset, should be regarded as tuition for, in substance, it served the same function as tuition.2 15 Applying that test here, the conclusion is inescapable that six hundred and forty dollars of taxpayers' payment to the Society was non-deductible tuition. The taxpayers' two children obtained a year of education in the Society's religiously-oriented school, as desired by taxpayers. The cost to the Society of providing that service was at least six hundred and forty dollars. In effect, if not in form or by design, taxpayers paid this cost. 16 Affirmed. * Of the Ninth Circuit, sitting by designation 1 Section 262 of the Code provides that "(e)xcept as otherwise expressly provided in this chapter, no deduction shall be allowed for personal, living, or family expenses." Tuition paid for the education of the children of a taxpayer is a family expense, not a charitable contribution to the educating institution. Channing v. 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Despite the many neuroimaging studies that suggest gray matter volume reductions in schizophrenia, there is no compelling postmortem evidence to suggest neuronal loss, nor is there a distinctive or specific signature of gray matter abnormalities in schizophrenia. Instead, there appears to be no observed focal lesion that characterizes gray matter pathology in schizophrenia. This state of affairs has led to a movement away from conceptualizing schizophrenia as resulting from a single lesion, to conceptualizing schizophrenia as arising from abnormal communication between brain regions. Abnormal connectivity between brain regions is not a new concept, as Bleuler (1911) postulated such abnormalities. What is new, however, is an appreciation that brain regions that are not spatially proximal may be connected functionally into neural networks and that to understand altered neural connectivity we need to have a better understanding of the connections between brain regions which are made possible by white matter, the main infrastructure in the brain that makes possible long distance communication among neurons. Accordingly, a focus on white matter connections in the brain has become increasingly of interest in schizophrenia, particularly in the areas of imaging studies, postmortem studies, and new animal models of schizophrenia. This special issue focuses on schizophrenia and oligodendrocytes, the latter a class of neuroglia that give rise to myelin. The primary aim of this special issue is to understand and to clarify further white matter pathology in schizophrenia and how it may contribute to disconnectivity among brain regions, which results in the observed cognitive, behavioral, and clinical symptoms in this disorder. The first paper begins with a description of the theory that schizophrenia reflects a disorder of connectivity. The proposition put forth is that schizophrenia is, at least in part, the result of abnormal communication between brain regions that may be spatially distant but nonetheless functionally connected. As white matter is the main infrastructure for brain connectivity, this paper focuses on the use of a magnetic resonance imaging (MRI) technique called diffusion tensor imaging (DTI) to gain insight into the role of white matter abnormalities in schizophrenia. This paper also focuses on the functional implications of white matter abnormalities, particularly with regard to the possible role of myelin in modulating the transmission velocity of neural discharges. The paper ends with a speculative hypothesis about the relationship between gray matter and white matter abnormalities in schizophrenia. The second paper also focuses on altered neuronal connectivity. Here, however, the focus is on altered connectivity and impaired myelination in a postmortem study of schizophrenia and normal controls, where electron microscopy is used to study myelinated fibers and oligodendrocytes. This morphometric study examines myelinated fibers in the prefrontal cortex in both gray and white matter. Six types of abnormal fibers and ultrastructural alterations are described in the schizophrenia sample. The study reveals increased pathological fibers in gray matter in both young and elderly patients that are associated with positive symptoms, while in elderly patients, the frequency of pathological fibers in white matter is increased and these changes are associated with more negative symptoms. The key implication here is that altered myelinated fibers in white matter in schizophrenia likely follow alterations of myelinated fibers in gray matter that occur earlier in the course of illness. The third paper is a postmortem study on age-related changes in the number of oligodendrocytes in prefrontal cortex in schizophrenia, bipolar disorder and major depression. The study reveals an age-related increase in numerical density of oligodendrocytes in layer VI and adjacent white matter of BA9 and 10 that is seen only in normal controls and not in schizophrenia or in mood disorders. The absence of the normal age-related increase in oligodendrocytes in patients suggests that this aspect of normal brain development is dysregulated in schizophrenia and in mood disorders, and confirms postmortem and imaging data, further highlighting the fact that the absence of a normal age-related increase in oligodendrocytes is a key shared feature in these psychiatric populations. The fourth paper reviews white matter abnormalities in schizophrenia, including imaging and postmortem findings on the disconnectivity theory of schizophrenia, a major theme that underlies the three previous papers in this special issue. This paper then reviews some of the white matter diseases that commonly lead to psychotic symptoms and then reviews transgenic and mutant mouse models of schizophrenia. More specifically, genetic mouse models such as Plp1 transgenic mice and mutant mice heterogeneous for either NRG1 or its receptor erbB4, as well as Nogo-A- deficient mice are reviewed. This is followed by a focus on the myelin toxicity model of mice fed cuprizone. In the early stage (the second and third week) of the cuprizone feeding, mice show higher dopamine levels and lower norepinephrine levels in their prefrontal cortex, along with behavioral changes indicative of increased CNS activity. In the late stage (weeks 4–6), when demyelination and oligodendrocyte loss are obvious, mice display cognitive deficits as well as deficits in social interactions that are reminiscent of social withdrawal seen in patients with schizophrenia. Importantly, this cuprizone-feeding mouse may be used as an in vivo platform for psychopharmacological studies testing the effects of antipsychotic drugs on white matter alterations and associated behavioral changes, in addition to providing a new animal model of schizophrenia. The fifth and final paper of this special issue focuses on abnormal behavior and microstructural changes in juvenile mice that are repeatedly exposed to nonneurotoxic levels of amphetamine. These mice are compared with nonamphetamine treated mice. Oligodendrocyte numbers and three proteins expressed in mature oligodendrocytes are investigated in these young male mice at sacrifice. Treated mice showed higher locomotion and impaired spatial working memory, along with lowered Noga-A and GST-pi proteins, and lower MBP proteins, in frontal cortex and hippocampus and fewer mature oligodendrocytes in frontal cortex and corpus callosum, as well as lower MBP staining in frontal cortex, corpus callosum, and hippocampus, than untreated mice. These differences suggest that in wild-type mice late developing white matter is vulnerable to amphetamine and may lead to compromised white matter with increased dopamine in specific brain regions. Not only do these findings explain well the increased locomotion and impaired spatial working memory observed in amphetamine-treated mice, but also may serve as a principle for the amphetamine-treated mouse being used as a novel animal model of schizophrenia. To summarize, compared to the focus on gray matter and neurons, research on white matter and myelin/glial has been sparse in schizophrenia. In this special issue, neuroimaging findings are presented that suggest that myelin abnormalities in schizophrenia may underlie the white matter abnormalities observed using diffusion tensor imaging techniques. Data from postmortem studies also confirm the presence of oligodendrocyte and myelin abnormalities in schizophrenia as well as mood disorders. Oligodendrocytes provide protection and improve communication between brain regions and the oligodendroglia-producing myelin is approaching maturity in the same time frame as the onset of symptoms. Further, normal age-related white matter increases and related gray matter changes that likely predate the white matter changes are dysregulated in schizophrenia and may contribute to the clinical symptomatology of the disease. Finally, the recently developed animal models are not only of help in understanding connectivity abnormalities in schizophrenia but also provide novel platforms to test novel therapeutic approaches for schizophrenic patients. Treatments specifically targeting white matter, for example, may improve on the efficacy of those treatments currently in use.

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1. Introduction {#sec1-ijerph-12-09692} =============== South Africa is a water stressed country due to the low average received rainfall (465 mm), which is below the global average of 860 mm \[[@B1-ijerph-12-09692]\]. Demand for this important scarce resource is expected to increase due to rapid industrial development, increasing human population, *per capita* consumption increases and the resulting impact of human activities on the environment \[[@B2-ijerph-12-09692],[@B3-ijerph-12-09692]\]. High water demand and consumption also leads to increases in the volume of wastewater generated \[[@B4-ijerph-12-09692]\]. The availability of good quality water is of paramount importance bringing to the fore the consequence of contamination of water bodies with pathogenic microorganisms \[[@B5-ijerph-12-09692]\]. Water bodies such as rivers are subject to dramatic changes in microbial and physicochemical qualities as a result of a variety of anthropogenic activities in the watershed. These changes are caused by discharges of municipal raw waters or treated effluent at a specific point source into the receiving surface waters \[[@B6-ijerph-12-09692],[@B7-ijerph-12-09692]\]. Point-source pollution problems will not only increase treatment costs considerably, but may also introduce a wide range of pathogens and harmful chemicals into surface waters that may be supplied to many rural and urban communities, thus resulting in incidences of waterborne diseases with far reaching socio-economic implications \[[@B8-ijerph-12-09692],[@B9-ijerph-12-09692]\]. Although a vast majority of microorganisms present in wastewater are not pathogenic \[[@B10-ijerph-12-09692]\], some pathogenic bacteria possibly originating from discharge of inadequately treated wastewater effluent have been implicated in the outbreak of waterborne diseases over the years \[[@B11-ijerph-12-09692]\]. *Salmonella* spp. are important Gram-negative bacilli, which infect both humans and animals causing a wide range of diseases such as gastroenteritis, typhoid fever, osteomyelitis, septicemia and meningitis. This genus comprises of over 2000 recognized serotypes and is divided into two species namely *S. bongori* and *S. enterica*. *Salmonella enterica* consist of six subspecies, namely *enterica, arizonae, salamae, diarizonae, houtenae and indica* \[[@B12-ijerph-12-09692],[@B13-ijerph-12-09692]\]. It is estimated that 93.8 million cases of gastroenteritis due to *Salmonella* spp. occur globally each year, with some 155,000 deaths \[[@B14-ijerph-12-09692]\]. This high number of infections emphasizes the importance of this intracellular pathogen and indicates a considerable worldwide disease burden. Mortality rate of *Salmonella* infections is a health problem mainly in developing countries \[[@B15-ijerph-12-09692]\], while morbidity due to acute *Salmonella* infection can also have socio-economic impacts in developed countries \[[@B16-ijerph-12-09692]\]. Added to this disease burden are the complications arising from the inefficacy and failures of antimicrobial chemotherapies applied in clinical practice to remedy these diseases. Bacterial resistance to antibiotics has increased globally in recent years and poses threats to human health \[[@B17-ijerph-12-09692]\]. Antimicrobial resistance in *Salmonella* has been associated with an increase in the number of adverse events following infection such as higher levels of hospitalization, longer illness, and higher risk of invasive illness as well as treatment failures \[[@B18-ijerph-12-09692]\]. The World Health Organization (WHO), the European Commission and the United States Centre for Disease Control and prevention (CDC) have recognized the importance of studying the emergence of resistance genes as well as the need for control strategies \[[@B19-ijerph-12-09692]\]. In most countries, the microbial quality of final treated effluent is estimated based on the level of indicator organisms present \[[@B20-ijerph-12-09692],[@B21-ijerph-12-09692]\]. However, several studies have shown that the presence of indicator organisms does not always correlate with the presence of pathogens \[[@B22-ijerph-12-09692],[@B23-ijerph-12-09692]\]. This study therefore aimed to determine the prevalence of antibiotic resistant and virulent *Salmonella* spp. in treated effluents and receiving surface water of two wastewater treatment plants located in Durban, South Africa. 2. Materials and Methods {#sec2-ijerph-12-09692} ======================== 2.1. Description of Sampling Site and Sample Collection {#sec2dot1-ijerph-12-09692} ------------------------------------------------------- Two wastewater treatment plants, namely the Northern Wastewater Treatment Works (NWWTW) and the New Germany Wastewater Treatment Plant (NGWTP) located in Durban, South Africa were sampled and studied. Details of these treatment plants can be found elsewhere \[[@B24-ijerph-12-09692]\]. The NWWTW and NGWTP discharge their final effluents into the Umgeni and Aller rivers, respectively. Water samples were collected monthly from both wastewater treatment plants at the clarifier before chlorination (B.C), discharge point after chlorination (D.P), 500 meters upstream (U.S) and 500 meters downstream (D.S) of the discharge point between March 2012 and February 2013. Samples were collected in 5 L plastic containers disinfected 24 h prior to collection by rinsing with deionized water and soaking in 70% ethanol. During collection of samples, the containers were rinsed with the sampled water before filling (at a depth of approximately one metre at each sampling point) to three-quarter of the container leaving space to allow for proper mixing. The collected samples were placed in ice packs, transported to the laboratory at the Department of Microbiology, University of KwaZulu-Natal (Westville) and processed within 24 h of collection. 2.2. Physico-Chemical Analysis of Water Samples {#sec2dot2-ijerph-12-09692} ----------------------------------------------- Temperature of the water samples was measured on site with a mercury thermometer; the pH was determined using a pH meter (Beckman, CA, USA) while turbidity was measured with a turbidimeter (21000P, HACH, CO, USA). Biochemical oxygen demand (BOD~5~) was determined using the LDC 101 probe with an HQ40d multimeter (HACH) after incubation for a period of 5 days according to the manufacturer's instructions. Chemical oxygen demand (COD) was measured with a Nova 60 spectroquant (Merck, Darmstadt, Germany) according to the manufacturer's instructions. Conductivity and total dissolved solids (TDS) were measured using the CDC 401 probe and HQ40d multimeter (HACH Colorado, USA). Nitrate (NO~3~) and Phosphate (PO~4~) concentrations were determined at Aquatico Laboratories (Pretoria, South Africa) using 50 mL of water samples. 2.3. Microbial Profile of Treated Wastewater Effluent and Receiving River {#sec2dot3-ijerph-12-09692} ------------------------------------------------------------------------- In order to avoid overestimation, *Salmonella* present in the water samples was quantified via a standard membrane filtration technique as previously described \[[@B3-ijerph-12-09692]\]. Ten-fold serial dilutions of the water samples were made and membrane filter (Pall Corporation, MI, USA) of 0.45 µm pore size and 47 mm diameter was used to concentrate 50 mL of appropriately diluted water sample. The membrane filter was then placed on the surface of *Salmonella-Shigella* (SS) agar and incubated aerobically at 37 °C for 18 to 24 h to enumerate *Salmonella* spp. Colourless colonies with or without black center depending on the production of hydrogen sulphide were enumerated as presumptive *Salmonella* spp. \[[@B25-ijerph-12-09692]\]. The organisms are referred to as presumptive *Salmonella* because certain bacteria also exhibit the same characteristics as *Salmonella* on SS agar. 2.4. Enrichment and Isolation of Salmonella spp. from Water Samples {#sec2dot4-ijerph-12-09692} ------------------------------------------------------------------- Isolation of *Salmonella* spp. from the water samples was done by a previously described enrichment method \[[@B26-ijerph-12-09692]\] with modifications. Thoroughly mixed water samples (25 mL) were added to 250 mL of sterile buffered peptone water and incubated at 37 °C for 18 to 24 h with shaking at 230 rpm. Thereafter, 1 mL of the pre-enrichment solution was appropriately diluted in 9 mL of sterile Rappaport-Vassiliadis soy broth (RVS) (Oxoid, Basingstoke, UK) depending on the turbidity of the water sample used in the pre-enrichment and incubated at 42 °C for 24 to 48 h with shaking at 230 rpm. One hundred microliters (100 µL) of the appropriately diluted RVS broth was spread-plated on *Salmonella* chromogenic agar (Oxoid) in duplicate and incubated aerobically at 37 °C for 18 to 24 h. Colonies with purple color were isolated and purified on fresh nutrient agar plates and subjected to further identification using biochemical tests and molecular method. ### 2.4.1. Biochemical Screening of Presumptive *Salmonella* spp. {#sec2dot4dot1-ijerph-12-09692} Triple sugar iron (TSI), Simmons citrate, lysine iron agar (LIA) and urea agar (Oxoid) slants were prepared according to the manufacturer's instructions and inoculated with a 24 h nutrient agar-grown culture of the isolates incubated at 37 °C. The surface of the agar slant was inoculated using a sterile inoculating loop while a stab was made at the center of the slant using a sterile inoculating needle. The tubes were then incubated under aerobic conditions at 37 °C for 24 to 48 h. Tubes exhibiting alkaline slant and acidic butt with H~2~S production on TSI slants, purple color in butt of LIA tube, blue color development on slant of citrate agar and no color change on urea indicated positive results for *Salmonella* spp. Isolates showing biochemical reaction consistent with *Salmonella* spp. were further confirmed using molecular based method. ### 2.4.2. Molecular Confirmation of Presumptive *Salmonella* spp. {#sec2dot4dot2-ijerph-12-09692} Template DNA was prepared from 24 h freshly grown cultures of the isolates on nutrient agar using the boiling method as previously described \[[@B27-ijerph-12-09692]\] with modifications. Well isolated colonies (3 to 5) were suspended in 70 µL of sterile deionized water, boiled in a water bath at 100 °C for 10 min and cooled on ice for a further 5 min. Thereafter, the suspension was centrifuged at 13,000 rpm in a micro-centrifuge (Eppendorf 5415D, Hamburg, Germany) for 5 min. The supernatant (50 µl) was carefully transferred to a sterile eppendorf tube and used as a template in the PCR assay. *Salmonella* spp. was confirmed by the amplification of the *inv*A gene using the primers F-5′-TGC-CTA-CAA-GCA-TGA-AAT-GG-3′ and R-5′-AAA-CTG-GAC-CAC-GGT-TGA-CAA-3′ \[[@B28-ijerph-12-09692]\]. The PCR mixture contained: 1 × PCR reaction buffer, 1 mM of MgCl~2~, 200 µM of each dNTPs, 0.5 µM of each primer, 2 U of *Taq* polymerase (Supertherm, Inqaba, Pretoria, South Africa) and 2 µL of template DNA in a final volume of 25 μL. Amplification was performed in a thermocycler (Bio-Rad T100, Singapore) with a temperature regime of 2 min at 94 °C for initial denaturation followed by 35 cycles of 94 °C for 1 min, annealing at 58 °C for 1 min, extension at 72 °C for 1 min with a final extension step at 72 °C for 5 min. PCR mixture with 2 µL of molecular grade water was used as negative control while *S. typhimurium* ATCC 13317 was used as a positive control. The PCR products were examined by electrophoresis in a 1.5% (w/v) agarose gel at 60 V for 90 min in 1% TAE buffer and visualized via UV illumination (Syngene Cambridge, UK) after staining in 1 mg/mL ethidium bromide for 15 min. ### 2.4.3. Virulence Gene Detection {#sec2dot4dot3-ijerph-12-09692} The isolates were evaluated for the presence of virulence genes in *Salmonella* pathogenicity island (SPI) 2 through 5 using the primers shown in [Table 1](#ijerph-12-09692-t001){ref-type="table"} \[[@B29-ijerph-12-09692],[@B30-ijerph-12-09692]\]. The presence of *mis*L and *orf*L virulence genes was confirmed using duplex PCR method developed in this study, while *spi*C and *pip*D were detected in separate single PCR assay. The reaction was done in a 25 μL reaction volume consisting of 1 × buffer, 1 mM MgCl~2~, 0.2 mM dNTPs, 0.5 μM of each primer, 2 μL of template DNA and 2.5 U of *Taq* polymerase (Supertherm). Amplification was carried out in a thermocycler (Bio-Rad T100) using a temperature program consisting of initial denaturation of 94 °C for 2 min, followed by 35 cycles of 94 °C for 1 min, 1 min at the respective annealing temperature of various primers ([Table 1](#ijerph-12-09692-t001){ref-type="table"}), 72 °C for 1 min with a final extension at 72 °C for 5 min. PCR mixture with 2 µL of molecular grade water was used as negative control while *S. typhimurium* ATCC 13317 was used as a positive control. The amplicons were examined by electrophoresis in a 1.5% agarose gel at 60 V for 90 min, stained in 1 mg/mL ethidium bromide for 5 min and viewed under UV light (Syngene Cambridge, UK). ### 2.4.4. Antibiotic Resistance Determination {#sec2dot4dot4-ijerph-12-09692} Antibiotic susceptibility of the isolates was determined using the Kirby-Bauer disk diffusion method. The isolates were screened against a predetermined and commercially available panel of 20 antibiotics (Oxoid), belonging to six classes. Fresh culture were grown overnight in Mueller-Hinton broth and standardized to 0.5 McFarland by diluting with sterile Mueller-Hinton broth until an optical density of 0.08 to 0.1 was obtained on a spectrophotometer (Shimadzu UV-1800, Kyoto, Japan) at 625 nm. The standardized culture of the isolates were inoculated onto Mueller-Hinton agar using sterile swabs for confluence growth and allowed to dry for 10 min. Thereafter, appropriate antibiotic disks were placed at equidistance on the surface of the agar plates with a sterile forceps and incubated at 37 °C for 24 h. The diameter of the zone of inhibition was measured to the nearest millimeter and interpreted using charts recommended by the CLSI \[[@B31-ijerph-12-09692]\]. 2.5. Statistical Analysis {#sec2dot5-ijerph-12-09692} ------------------------- Mean values of results and standard deviation were calculated using Microsoft Excel 2010 edition. Pearson's correlation was determined using the SPSS 21.0 for windows program (SPSS, Inc. WA, USA) after normalization of skewed datasets. Correlations were considered statistically significant at *p* values of \<0.05. ijerph-12-09692-t001_Table 1 ###### Primer used for the detection of virulence genes in *Salmonella* spp. recovered from treated wastewater effluent and receiving surface waters. Gene Target Oligonucleotide Sequence (5ʹ-3ʹ) Amplicon Size (bp) Annealing Temperatures (°C) ------------------------ ---------------------------------- -------------------- ----------------------------- *spi*C f-CCTGGATAATGACTATTGAT 309 54 r-AGTTTATGGTGATTGCGTAT *pip*D f-CGGCGATTCATGACTTTGAT 400 56 r-CGTTATCATTCGGATCGTAA *mis*L f-GTCGGCGAATGCCGCGAATA 550 60 r-GCGCTGTTAACGCTAATAGT *orf*L f-GGAGTATCGATAAAGATGTT 350 60 r-CGTTATCATTCGGATCGTAA 3. Results {#sec3-ijerph-12-09692} ========== 3.1. Physicochemical Profile of Water Samples {#sec3dot1-ijerph-12-09692} --------------------------------------------- The physicochemical parameters of treated effluent at the NWWTP and NGWTP and the receiving surface waters are shown in [Table 2](#ijerph-12-09692-t002){ref-type="table"} and [Table 3](#ijerph-12-09692-t003){ref-type="table"}. Temperature was stable across all sampled points in each month but seasonal variations were observed. The lowest temperature recorded was 12 °C, while the highest temperature of 27 °C. The pH was stable at all sampled points in each month but varied throughout the duration of the study. It ranged from 6.41 (at the U.S in September) to 7.88 (at the D.P in February) at the NWWTW while at the NGWTP, the pH ranged from 6.30 (at the U.S in July) to 8.00 (at the D.S in February). Turbidity values recorded varied across all sampled point and month with no significant decrease in turbidity obtained at the D.P compared to B.C point ([Table 2](#ijerph-12-09692-t002){ref-type="table"}). At the NGWTP, recorded values ranged from 1.42 NTU at D.P in April to 40.40 NTU at U.S in August. The COD values varied highly throughout the study period ranging from 22.33 to 313 mg/L at U.S at the NGWTP. Some levels of decrease at the D.P was observed only in the months of April (36.5%), May (21.11%) and June (55.6%) but increased in the other months. Recorded COD values also varied highly at the NWWTW and ranged from \<10 mg/L in March and May (at the D.P) to 312.44 mg/L in July (at the D.S). Some level of decrease in COD was recorded at the D.P in May, August, October and December but increased in other months especially in March and May from \<10 mg/L to 309 mg/L. Stable BOD~5~ values were observed across each sampled point in each month ranging from 1.03 mg/L to 9.42 mg/L at the NWWTW. The values of BOD~5~ at the D.S were higher than values recorded at other sampled points and ranged from 3.58 mg/L to 7.74 mg/L. At the NGWTP, the BOD~5~ was stable across all sampled points in each month but varied throughout the study period ranging from 2.20 mg/L to 11.04 mg/L. ijerph-12-09692-t002_Table 2 ###### Physicochemical parameters of wastewater effluent from Northern wastewater treatment works and the receiving river from March 2012 to February 2013. MONTH Sampled Point Parameter (Mean ± SD) ------- --------------- ----------------------- ------------- --------------- ---------------- ------------- ------ ------- U.S 26.00 ± 0.00 7.25 ± 0.09 16.67 ± 0.38 161.33 ± 4.37 5.62 ± 1.01 1.06 0.05 MAR B.C 26.00 ± 0.00 7.11 ± 0.05 7.91 ± 0.33 104.78 ± 13.73 2.23 ± 0.36 0.06 3.51 D.P 25.00 ± 0.00 7.36 ± 0.07 23.40 ± 12.13 \<10 ± 0.00 5.13 ± 0.18 0.06 3.47 D.S 26.00 ± 0.00 7.24 ± 0.06 15.27 ± 0.12 309.33 ± 0.58 5.62 ± 0.24 0.06 0.16 U.S 21.00 ± 0.00 7.43 ± 0.12 19.7 ± 0.00 304.33 ± 2.08 8.49 ± 0.47 0.86 0.23 B.C 22.00 ± 0.00 7.67 ± 0.06 56.53 ± 0.12 229.33 ± 9.71 3.30 ± 0.97 0.05 2.11 APR D.P 22.00 ± 0.00 7.40 ± 0.10 76.43 ± 0.29 311.11 ± 2.01 3.44 ± 0.67 0.05 2.10 D.S 21.00 ± 0.00 7.63 ± 0.06 14.80 ± 0.00 151.00 ± 0.00 6.33 ± 0.21 0.32 0.38 U.S 21.00 ± 0.00 6.91 ± 0.04 12.80 ± 0.00 20.22 ± 1.71 4.29 ± 0.79 1.03 1.08 B.C 22.00 ± 0.00 7.08 ± 0.03 19.60 ± 0.00 38.22 ± 11.55 1.03 ± 0.19 0.16 2.72 MAY D.P 21.00 ± 0.00 7.28 ± 0.02 13.80 ± 0.17 \<10 ± 0.00 3.25 ± 0.17 0.20 2.34 D.S 22.00 ± 0.00 7.13 ± 0.03 12.90 ± 0.00 309.11 ± 1.71 5.68 ± 0.30 1.72 0.82 U.S 13.00 ± 0.00 7.65 ± 0.01 9.57 ± 0.01 112.89 ± 3.02 9.42 ± 0.15 2.40 2.17 B.C 13.00 ± 0.00 7.37 ± 0.00 11.27 ± 0.31 300.00 ± 8.65 4.36 ± 0.16 5.98 12.38 JUN D.P 12.00 ± 0.00 7.35 ± 0.01 8.92 ± 0.06 110.00 ± 3.06 4.54 ± 0.12 3.92 9.02 D.S 14.00 ± 0.00 7.84 ± 0.01 14.37 ± 0.21 88.78 ± 2.41 7.74 ± 0.31 1.78 1.44 U.S 15.00 ± 0.00 7.54 ± 0.01 13.27 ± 0.15 311.00 ± 1.00 5.76 ± 1.03 0.79 0.03 B.C 16.00 ± 0.00 7.48 ± 0.01 19.33 ± 0.06 114.78 ± 11.65 2.56 ± 0.58 1.00 1.83 JUL D.P 15.00 ± 0.00 7.70 ± 0.00 23.07 ± 0.38 290.67 ± 0.88 3.12 ± 0.62 0.89 0.75 D.S 15.00 ± 0.00 7.87 ± 0.02 22.87 ± 0.12 312.44 ± 0.38 4.69 ± 0.23 1.35 0.09 U.S 20.00 ± 0.00 7.12 ± 0.03 28.73 ± 0.06 105.89 ± 3.86 2.80 ± 0.57 0.06 0.12 B.C 21.00 ± 0.00 6.85 ± 0.11 56.37 ± 0.35 310.11 ± 0.69 1.51 ± 1.09 0.00 0.00 AUG D.P 19.00 ± 0.00 7.09 ± 0.4 65.53 ± 0.57 182.78 ± 2.27 1.59 ± 0.84 0.06 1.99 D.S 20.00 ± 0.00 7.26 ± 0.02 20.77 ± 0.06 309.56 ± 2.14 3.98 ± 0.65 0.00 0.00 U.S 20.00 ± 0.00 6.41 ± 0.05 10.67 ± 0.06 55.56 ± 0.51 3.73 ± 0.53 0.17 0.10 B.C 22.00 ± 0.00 6.76 ± 0.02 20.73 ± 0.06 308.67 ± 0.88 1.51 ± 0.76 0.15 1.70 SEPT D.P 20.00 ± 0.00 6.82 ± 0.04 19.27 ± 0.21 308.44 ± 1.26 2.38 ± 1.10 0.08 1.90 D.S 20.00 ± 0.00 6.52 ± 0.02 11.50 ± 0.10 139.67 ± 1.73 3.92 ± 0.78 1.47 0.16 U.S 24.00 ± 0.00 7.02 ± 0.01 17.07 ± 0.12 195.22 ± 3.98 3.32 ± 0.78 1.02 0.37 B.C 22.00 ± 0.00 6.60 ± 0.06 30.53 ± 0.23 306.89 ± 1.84 3.23 ± 1.40 0.88 1.88 OCT D.P 23.00 ± 0.00 6.75 ± 0.05 28.50 ± 0.00 109.89 ± 2.80 3.88 ± 0.75 0.82 1.57 D.S 24.00 ± 0.00 6.91 ± 0.01 29.03 ± 0.06 148.00 ± 0.33 3.58 ± 0.98 1.30 0.42 U.S 21.00 ± 0.00 6.86 ± 0.01 21.33 ± 0.76 241.78 ± 21.56 4.24 ± 0.98 0.97 0.64 NOV B.C 22.00 ± 0.00 6.79 ± 0.01 39.13 ± 0.40 123.78 ± 6.91 3.56 ± 0.92 0.28 3.53 D.P 23.00 ± 0.00 6.68 ± 0.03 48.53 ± 0.55 287.22 ± 14.25 3.26 ± 0.88 0.31 3.69 D.S 23.00 ± 0.00 6.72 ± 0.05 14.10 ± 0.46 246.11 ± 14.84 3.87 ± 0.81 3.38 0.45 U.S 22.00 ± 0.00 6.85 ± 0.01 12.20 ± 0.26 274.33 ± 4.41 3.65 ± 0.78 0.39 0.03 B.C 25.00 ± 0.00 6.78 ± 0.03 36.13 ± 0.40 170.78 ± 3.79 3.29 ± 0.96 0.24 1.82 DEC D.P 21.00 ± 0.00 6.69 ± 0.01 31.77 ± 0.23 153.89 ± 0.19 3.52 ± 0.77 0.77 1.37 D.S 22.00 ± 0.00 6.64 ± 0.02 10.33 ± 0.41 205.33 ± 4.98 4.01 ± 0.79 1.95 0.35 U.S 24.00 ± 0.00 7.04 ± 0.01 11.40 ± 0.26 299.22 ± 1.07 3.76 ± 0.67 0.58 0.23 JAN B.C 24.00 ± 0.00 6.84 ± 0.01 12.67 ± 0.15 \<10 ± 0.00 3.68 ± 0.94 0.02 4.70 D.P 23.00 ± 0.00 6.87 ± 0.03 32.67 ± 0.81 303.67 ± 0.33 3.72 ± 0.95 0.02 4.69 D.S 24.00 ± 0.00 6.92 ± 0.02 8.72 ± 0.04 150.11 ± 3.56 3.74 ± 0.66 1.22 0.12 U.S 25.00 ± 0.00 7.41 ± 0.01 6.37 ± 0.02 308.89 ± 1.02 3.26 ± 0.39 0.22 0.03 FEB B.C 25.00 ± 0.00 7.80 ± 0.01 40.37 ± 0.21 295.67 ± 4.73 1.82 ± 1.12 0.01 3.51 D.P 25.00 ± 0.00 7.88 ± 0.01 44.07 ± 0.25 309.33 ± 0.58 2.81 ± 0.86 0.02 2.71 D.S 27.00 ± 0.00 7.77 ± 0.01 5.94 ± 0.10 254.78 ± 5.39 4.01 ± 0.80 0.16 0.53 ijerph-12-09692-t003_Table 3 ###### Physicochemical parameters of wastewater effluent from New Germany wastewater treatment works and the receiving river from March 2012 to February 2013. MONTH Sampled Point Parameter (Mean ± SD) ---------- --------------- ----------------------- ------------- -------------- ---------------- -------------- --------- --------- U.S 26.00 ± 0.00 7.52 ± 0.09 5.15 ± 0.05 313.89 ± 0.19 7.79 ± 0.83 1.06 0.05 MAR B.C 26.00 ± 0.00 7.12 ± 0.21 6.65 ± 0.23 153.67 ± 11.46 2.20 ± 0.13 1.81 0.90 D.P 26.00 ± 0.00 7.18 ± 0.12 5.71 ± 0.59 239.00 ± 10.00 3.12 ± 0.27 1.40 0.80 D.S 26.00 ± 0.00 7.51 ± 0.09 7.32 ± 0.33 141.33 ± 11.06 4.97 ± 0.59 2.14 0.05 U.S 18.00 ± 0.00 7.08 ± 0.02 8.23 ± 0.00 104.22 ± 2.83 8.49 ± 0.47 1.29 0.17 B.C 20.00 ± 0.00 7.04 ± 0.04 1.52 ± 0.00 202.78 ± 9.10 3.30 ± 0.97 2.81 1.09 APR D.P 19.00 ± 0.00 6.82 ± 0.01 1.42 ± 0.02 179.67 ± 1.20 3.44 ± 0.67 2.35 0.86 D.S 20.00 ± 0.00 7.05 ± 0.04 17.00 ± 0.00 114.00 ± 1.73 6.33 ± 0.21 0.82 0.28 U.S 16.00 ± 0.00 6.42 ± 0.05 3.18 ± 0.00 298.67 ± 0.33 11.04 ± 0.97 0.84 \<0.025 B.C 19.00 ± 0.00 6.91 ± 0.09 28.70 ± .00 312.22 ± 0.69 3.15 ± 0.25 \<0.057 0.05 MAY D.P 14.00 ± 0.00 7.02 ± 0.01 30.30 ± 0.00 246.33 ± 3.06 4.72 ± 0.16 \<0.057 \<0.025 D.S 19.00 ± 0.00 7.10 ± 0.00 17.80 ± 0.00 311.89 ± 2.22 9.67 ± 0.55 0.51 \<0.025 U.S 16.00 ± 0.00 7.93 ± 0.01 9.02 ± 0.12 22.33 ± 3.79 10.80 ± 0.41 2.38 0.66 B.C 18.00 ± 0.00 7.62 ± 0.01 9.63 ± 0.03 310.00 ± 1.73 4.19 ± 0.11 3.76 1.05 JUN D.P 14.00 ± 0.00 7.55 ± 0.01 10.63 ± 0.51 137.67 ± 9.87 5.03 ± 0.07 1.99 0.88 D.S 18.00 ± 0.00 7.83 ± 0.00 14.07 ± 0.12 73.33 ± 4.16 7.16 ± 1.57 2.15 0.33 U.S 14.00 ± 0.00 6.30 ± 0.01 2.44 ± 0.01 309.67 ± 2.19 5.27 ± 0.41 1.03 0.03 B.C 17.00 ± 0.00 6.53 ± 0.10 20.07 ± 0.12 193.67 ± 3.67 2.12 ± 0.17 0.47 0.03 JUL D.P 15.00 ± 0.00 6.89 ± 0.01 20.73 ± 0.15 308.67 ± 0.58 3.95 ± 0.34 0.57 0.09 D.S 17.00 ± 0.00 6.98 ± 0.02 16.10 ± 0.00 299.56 ± 4.62 9.42 ± 0.55 0.71 0.07 U.S 15.00 ± 0.00 7.12 ± 0.03 40.40 ± 0.36 207.56 ± 1.07 3.27 ± 1.02 2.29 0.59 B.C 17.00 ± 0.00 6.85 ± 0.11 19.73 ± 0.15 139.56 ± 1.02 3.62 ± 1.02 1.42 0.93 AUG D.P 12.00 ± 0.00 7.26 ± 0.02 16.80 ± 0.17 309.00 ± 1.33 4.68 ± 0.80 1.96 1.47 D.S 17.00 ± 0.00 7.09 ± 0.04 14.10 ± 0.10 311.78 ± 0.84 5.86 ± 1.57 0.80 1.89 U.S 20.00 ± 0.00 6.48 ± 0.02 15.83 ± 0.15 310.33 ± 0.58 4.06 ± 0.91 2.65 0.54 B.C 22.00 ± 0.00 6.75 ± 0.08 5.84 ± 0.01 98.22 ± 2.41 4.66 ± 0.84 8.22 0.58 **SEPT** D.P 20.00 ± 0.00 6.37 ± 0.03 16.33 ± 0.06 310.44 ± 1.17 4.49 ± 1.08 0.23 0.14 D.S 20.00 ± 0.00 6.59 ± 0.00 6.98 ± 0.02 189.89 ± 2.59 3.42 ± 0.47 0.72 0.17 U.S 17.00 ± 0.00 6.97 ± 0.02 3.68 ± 0.01 311.89 ± 0.84 4.46 ± 0.67 2.32 0.05 B.C 20.00 ± 0.00 6.91 ± 0.09 20.00 ± 0.10 35.67 ± 3.61 4.51 ± 0.84 1.23 1.12 **OCT** D.P 19.00 ± 0.00 6.85 ± 0.14 6.48 ± 0.04 54.11 ± 3.15 4.42 ± 0.72 1.64 1.26 D.S 20.00 ± 0.00 6.98 ± 0.03 5.10 ± 0.01 239.22 ± 4.81 4.79 ± 0.79 2.47 0.49 U.S 17.00 ± 0.00 7.12 ± 0.01 8.11 ± 0.06 306.78 ± 2.46 4.31 ± 0.78 2.11 0.02 **NOV** B.C 20.00 ± 0.00 6.82 ± 0.03 5.51 ± 0.08 69.11 ± 1.39 4.14 ± 0.61 0.50 3.22 D.P 18.00 ± 0.00 7.14 ± 0.04 29.43 ± 0.06 108.56 ± 3.24 4.22 ± 0.71 0.33 3.99 D.S 20.00 ± 0.00 7.16 ± 0.01 16.53 ± 0.23 257.56 ± 14.55 4.49 ± 0.81 1.27 1.70 U.S 20.00 ± 0.00 6.47 ± 0.01 32.10 ± 0.10 24.33 ± 2.08 3.38 ± 1.46 \<0.017 0.04 B.C 22.00 ± 0.00 6.55 ± 0.28 4.41 ± 0.30 93.33 ± 5.93 2.87 ± 0.81 0.11 0.13 **DEC** D.P 20.00 ± 0.00 6.45 ± 0.01 29.43 ± 0.06 300.44 ± 4.22 3.36 ± 0.92 \<0.017 0.14 D.S 22.00 ± 0.00 6.51 ± 0.04 28.10 ± 0.10 35.33 ± 3.84 4.17 ± 0.83 0.51 0.06 U.S 22.00 ± 0.00 6.71 ± 0.01 10.80 ± 0.00 80.33 ± 5.13 3.92 ± 0.69 1.21 \<0.08 **JAN** B.C 23.00 ± 0.00 6.59 ± 0.02 9.43 ± 0.04 111.11 ± 0.84 3.43 ± 1.09 \<0.017 0.01 D.P 22.00 ± 0.00 6.61 ± 0.02 9.29 ± 0.04 240.78 ± 3.75 3.75 ± 0.79 \<0.017 0.15 D.S 23.00 ± 0.00 6.73 ± 0.00 10.60 ± 0.00 44.56 ± 3.42 4.50 ± 1.27 0.84 0.13 U.S 21.00 ± 0.00 7.50 ± 0.05 8.83 ± 0.04 305.33 ± 0.58 3.04 ± 0.80 0.44 0.01 **FEB** B.C 24.00 ± 0.00 7.72 ± 0.03 3.93 ± 0.01 158.89 ± 4.72 3.97 ± 1.10 \<0.017 0.76 D.P 23.00 ± 0.00 7.87 ± 0.02 4.02 ± 0.03 283.44 ± 3.79 4.08 ± 1.04 \<0.017 1.16 D.S 24.00 ± 0.00 8.08 ± 0.00 5.80 ± 0.02 272.89 ± 14.82 4.85 ± 0.85 0.39 0.64 Values are averages of three replicates ± standard deviation. BOD values increased at the D.P during most of the study period but some level of decrease was recorded in September (17%) and October (9%). At the NWWTW, nitrate (NO~3~) values ranged from 0.00 mg/L at the B.C and D.S in August to 5.98 mg/L at the B.C in June, while phosphates (PO~4~) values ranged from 0 mg/L at the B.C and D.S in August to 12.38 mg/L. At the NGWTP, NO~3~ values ranged from \<0.06 mg/L (D.P, May) to 8.22 mg/L (B.C, September) while recorded PO~4~ values ranged from \<0.03 mg/L in May to 3.99 mg/L in November at the D.P ([Table 3](#ijerph-12-09692-t003){ref-type="table"}). 3.2. Enumeration of Presumptive Salmonella in Treated Wastewater Effluent and Receiving River {#sec3dot2-ijerph-12-09692} --------------------------------------------------------------------------------------------- The monthly variation of presumptive *Salmonella* spp. population in NWWTW and receiving Umgeni River are shown in [Figure 1](#ijerph-12-09692-f001){ref-type="fig"}a. At U.S, counts for presumptive *Salmonella* spp. ranged from 0.70 log·cfu/mL in October to 2.99 log·cfu/mL in March, while at the BC point, counts ranged from 1.05 to 3.51 log·cfu/mL in June and May, respectively. At the D.P there was no presumptive *Salmonella* recorded in October while the highest count of 3.29 log·cfu/mL was recorded in April. At the D.S, the counts ranged from 0.61 log·cfu/mL in February to 2.98 log·cfu/mL in April. The monthly variation of presumptive *Salmonella* spp. population in NGWTP is given in [Figure 1](#ijerph-12-09692-f001){ref-type="fig"}b. At the U.S, recorded values ranged from 1.04 log·cfu/mL in May to 3.02 log·cfu/mL in September. At the B.C point, the lowest count was recorded in August (0.98 log·cfu/mL) while the highest count was recorded in November (3.18 log·cfu/ml). No presumptive *Salmonella* was recorded at the D.P between June and August as well as December to February while the highest values was recorded in September (3.22 log·cfu/ml). At the D.S, the highest recorded value was obtained in the month of December (4.14 log·cfu/ml) while no *Salmonella* count was recorded in July, August, January and February. ###### (**a**) Monthly variation of presumptive *Salmonella* spp. at (**a**) NWWTP and (**b**) NGWTP and their receiving surface waters during the study period.   3.3. Statistical Analysis {#sec3dot3-ijerph-12-09692} ------------------------- [Table 4](#ijerph-12-09692-t004){ref-type="table"} and [Table 5](#ijerph-12-09692-t005){ref-type="table"} show the correlation matrices of selected physicochemical parameters with microbial counts. At the NWWTW ([Table 4](#ijerph-12-09692-t004){ref-type="table"}), there was a strong positive correlation between Presumptive *Salmonella* counts and turbidity (r = 0.501) as well as with PO~4~ (r = 0.431). NO~3~ also strongly correlated positively with temperature (0.430) and BOD~5~ (r = 0.462). However, a strong inverse correlation was seen between Turbidity and BOD5 (r = −0.378) as well as NO~3~ (r = 0.408). Presumptive *Salmonella* counts strongly correlated negatively with both BOD~5~ (r = −0.325) and NO~3~ (r = −0.349). At the NGWTP ([Table 5](#ijerph-12-09692-t005){ref-type="table"}), PO~4~ correlated significantly (*p* \> 0.05) with pH (r =0.302) but correlated strongly but negatively (*p* \< 0.01) with temperature (r = −0.430). ijerph-12-09692-t004_Table 4 ###### Correlation matrices of the physicochemical parameters at the NWWTW. Temp pH T COD BOD~5~ NO~3~ PO~4~ *Sal.* ^a^ ----------------- ------------ -------- ------------- -------- ------------ ----------- ------------ ------------ Temp (°C) 1 pH 0.140 1 T (ntu) 0.065 −0.028 1 COD (mg/L) −0.031 −0.225 −0.205 1 BOD~5~ (mg/L) 0.129 0.253 −0.378 \*\* 0.036 1 NO~3~ (mg/L) 0.430 \*\* 0.011 −0.408 \*\* 0.139 0.462 \*\* 1 PO~4~ (mg/L) 0.130 0.080 0.166 0.171 −0.169 0.118 1 *Sal.* (cfu/mL) −0.002 0.225 0.501 \*\* −0.078 −0.325 \* −0.349 \* 0.431 \*\* 1 **^a^**---*Salmonella* spp.; **\*\*** Correlation is significant at the 0.01 level (2-tailed); **\*** Correlation is significant at the 0.05 level (2-tailed). ijerph-12-09692-t005_Table 5 ###### Correlation matrices of physicochemical parameters at the NGWTP. Temp pH T COD BOD~5~ NO~3~ PO~4~ *Sal.* ^a^ ----------------- ----------------- -------------- -------- -------- -------- -------- -------- ------------ Temp (°C) 1 pH 0.094 1 T (ntu) −0.265 −0.148 1 COD (mg/L) 0.262 −0.010 −0.024 1 BOD~5~ (mg/L) −0.419 **\*\*** 0.264 −0.044 −0.195 1 NO~3~ (mg/L) −0.167 0.187 −0.240 0.046 0.151 1 PO~4~ (mg/L) −0.005 0.302 **\*** −0.158 0.213 −0.089 0.207 1 *Sal.* (cfu/mL) −0.002 −0.229 0.215 0.066 −0.246 −0.090 −0.161 1 **^a^**---*Salmonella* spp.; **\*\*** Correlation is significant at the 0.01 level (2-tailed); **\*** Correlation is significant at the 0.05 level (2-tailed). 3.4. Distribution of Confirmed Salmonella spp. Recovered from Treated Wastewater and Receiving Surface Waters {#sec3dot4-ijerph-12-09692} ------------------------------------------------------------------------------------------------------------- After enrichment and confirmation, a total of 200 of the recovered isolates were confirmed as *Salmonella* spp. The NGWTP and receiving surface water has the highest prevalence (93.5%) of *Salmonella* spp.. while only 6.5% of the isolates were recovered in treated effluent before chlorination at the NWWTW. At the NGWTP, 53 (26.5%) of the isolates were recovered in effluent sample before chlorination. Also, 55 (27.5%) isolates were recovered at the discharge point of the NGWTP with additional 27% and 12.5% recovered upstream and downstream of the receiving river of the treated final effluent from the NGTWP, respectively. 3.5. Virulence Gene Distribution and Antibiotic Resistance Profiles of the Positively Identified Salmonella spp. {#sec3dot5-ijerph-12-09692} ---------------------------------------------------------------------------------------------------------------- A representative gel of four virulence genes detected in *Salmonella* spp. is shown in [Figure 2](#ijerph-12-09692-f002){ref-type="fig"}. Analysis of the virulence gene signatures in recovered *Salmonella* spp. revealed that 93% harbored the *spi*C gene, 84% harbored the *misL* gene, and 87.5% harbored the *orf*L gene while 87% harbored *pip*D gene. Antibiotic resistance profile of the 200 confirmed isolates is shown in [Table 5](#ijerph-12-09692-t005){ref-type="table"}. {#ijerph-12-09692-f002} The isolates were highly susceptible to β-lactams, chloramphenicol, tetracycline, norfloxacin and trimethoprim-sulfamethoxazole (99% to 100%). Resistance was observed against sulfamethoxazole (100%), nalidixic acid (27%) and streptomycin (14%). Resistance to third generation β-lactams was also observed in 0.5% of the isolates. Intermediate resistance was observed against streptomycin (74%), nalidixic acid (44%) and fosfomycin (8.5%). Though some of the isolates recovered were susceptible to nalidixic acid (54%), high resistance to third generation quinolones such as ciprofloxacin, norflorxacin and nitrofurantoin were not recorded ([Table 6](#ijerph-12-09692-t006){ref-type="table"}). ijerph-12-09692-t006_Table 6 ###### Antibiotic resistance profile of *Salmonella* spp. recovered from wastewater treatment plants and receiving surface waters (n = 200). Class Antibiotics Content Resistant (%) Intermediate (%) Susceptible (%) ----------------- -------------------------------- --------------- --------------- ------------------ ----------------- β-lactams Cephalothin 30 μg 0 (0) 0 (0) 200 (100) Imipenem 10 μg 1 (0.5) 1 (0.5) 198 (99) Cefoxitin 30 μg 0 (0) 0 (0) 200 (100) Cefuroxime 30 μg 0 (0) 2 (1) 198 (99) Piperacillin 100 μg 1 (0.5) 3 (1.5) 196 (98) Ampicillin 10 μg 1 (0.5) 2 (1) 197 (98.5) Cefixime 5 μg 1 (0.5) 0 (0) 199 (99.5) Ceftazidime 30 μg 1 (0.5) 0 (0) 198 (99) Aztreonam 30 μg 0 (0) 0 (0) 200 (100) Aminoglycosides Gentamycin (10 μg) 0 (0) 0 (0) 200 (100) Amikacin 30 μg 0 (0) 1 (0.5) 199 (99.5) Streptomycin 10 μg 28 (14) 148 (74) 24 (12) Phenicols Chloramphenicol 30 μg 0 (0) 0 (0) 200 (100) Tetracyclines Tetracycline 30 μg 0 (0) 0 (0) 200 (100) Quinolones Ciprofloxacin 5 μg 0 (0) 1 (0.5) 199 (99.5) Norflaoxacin 10 μg 0 (0) 0 (0) 200 (100) Nalidixic acid 30 μg 54 (27) 89 (44.5) 57 (28.5) Nitrofurantoin 30 μg 0 (0) 17 (8.5) 183 (91.5) Sulfonamides Trimethorprim/Sulfamethoxazole 1.25/23.75 μg 0 (0) 0 (0) 200 (100) Sulfamethoxazole 5 μg 200 (100) 0 (0) 0 (0) 4. Discussion and Conclusions {#sec4-ijerph-12-09692} ============================= The impact of substandard quality effluent or untreated wastewater discharged into receiving water bodies can be detrimental to water availability and security. Wastewater management is the first barrier in a multi-barrier system to ensure safe drinking water, public health and environmental sustainability \[[@B32-ijerph-12-09692]\]. Physicochemical analysis of the wastewater gives an indication of the quality of effluents being discharge into the environment. The temperature regime varied depending on the season but was still within the acceptable limit of 25 °C \[[@B33-ijerph-12-09692]\] and did not pose any threat to the receiving watershed. The temperature is a very important parameter because of its effect on the chemical reaction and reaction rates, aquatic life and suitability of the water for beneficial uses. Increase in temperature can lead to the disruption of the ecosystem in the receiving watershed resulting in high mortality of aquatic life and encouraged growth of undesirable algae and wastewater fungi \[[@B34-ijerph-12-09692]\]. At both plants, the pH ranged between 6.41--8.08 and 6.37--7.88. The neutral to alkaline pH values recorded in this study are similar to previous reports \[[@B7-ijerph-12-09692]\]. Very low or high pH is known to be toxic to aquatic life and alters the solubility of chemicals in water \[[@B35-ijerph-12-09692]\]. The pH of most natural waters is in the range of 4−9 and the target limit set by the South African Department of Water Affairs is between 5.5 and 9.5 \[[@B33-ijerph-12-09692]\]. The pH of water can provide important information about many chemical and biological processes and provides indirect correlations to a number of different impairments in the wastewater treatment processes \[[@B36-ijerph-12-09692]\]. Changes in pH can also be indicative of industrial pollution, photosynthesis or the decomposition of organic matter by microorganisms \[[@B37-ijerph-12-09692]\]. Hence, the pH values recorded in this study fell within the acceptable range indicating that discharge of the treated wastewater may have no negative impact on the river water with respect to pH. The turbidity of the water samples in this study ranged between 1.42 NTU to 76.43 NTU and varied seasonally ([Table 2](#ijerph-12-09692-t002){ref-type="table"} and [Table 3](#ijerph-12-09692-t003){ref-type="table"}). Higher turbidity values recorded in some months at the D.P could be the result of storm runoff and inadequate treatment leading to the high variation in turbidity. This high variation has been reported in previous studies in the Eastern Cape province of South Africa \[[@B7-ijerph-12-09692]\]. The turbidity values recorded in this study was higher than the \<5 NTU set by the World Health Organization (WHO) for effluent discharge into the environment \[[@B7-ijerph-12-09692]\]. Turbidity is caused by small particles that may be organic or inorganic and can provide food and shelter for pathogens providing a possible explanation of its strong positive correlation with *Salmonella* count at the NWWTW ([Table 4](#ijerph-12-09692-t004){ref-type="table"}). If not removed, turbidity can promote the regrowth of pathogens in the final effluent of receiving water body into which the effluent is discharged \[[@B38-ijerph-12-09692]\]. Turbidity also limits the bactericidal effect of chlorine in the wastewater during disinfection \[[@B35-ijerph-12-09692]\] and may react with organic compounds in the water to form micro-contaminants such as trihalomethane (THM). THMs are carcinogenic and render the water unfit for recreational activities to consumers who may depend on the receiving water shed for such \[[@B9-ijerph-12-09692],[@B39-ijerph-12-09692]\]. Although the BOD~5~ values recorded was stable across each sampling point in each month, the values varied significantly in the course of the study ranging from 1.03 mg/L to 11.04 mg/L. There is no South African guideline for BOD~5~ levels in the final effluent of wastewater; however, the EU recommends a discharge limit of 3 to 6 mg/L for aquatic ecosystems \[[@B6-ijerph-12-09692]\]. Discharge of effluent high in BOD into water bodies would result in rapid depletion of dissolved oxygen leading to anoxic conditions, and consequent disruption of balance in the aquatic ecosystem \[[@B6-ijerph-12-09692]\]. On most occasions the recorded BOD~5~ values at the D.P were within the recommended EU limit. The COD of the water samples varied remarkably throughout the study period. High COD values were recorded at U.S while, the average recorded values (212 mg/L) at the D.P greatly exceeded the South African limit of 30 mg/L \[[@B33-ijerph-12-09692]\]. High levels of COD observed U.S could be attributed to runoff, agricultural activities and anthropogenic activities. Igbinosa and Okoh \[[@B7-ijerph-12-09692]\] reported a similar observation and attributed the increased COD to addition of organic and inorganic substances from the environment as well as organic contaminants entering the system from municipal sewage treatment plants or non-point sources of pollution. Higher average COD values varying from 512 to 698.11 mg/L were reported in a study on river quality in India and was attributed to the presence of inorganic chemicals in the wastewater of a nearby chemical industry \[[@B40-ijerph-12-09692]\]. Tertiary treatment of final sewage effluent with chlorine at the wastewater treatment plants (WWTPs) under investigation reduced the number of viable presumptive *Salmonella* spp. at the discharge point during the sampling period but failed to totally eliminate them ([Figure 1](#ijerph-12-09692-f001){ref-type="fig"}). Presumptive *Salmonella* spp. were also recovered downstream of the receiving rivers indicating the negative impact of the wastewater treatment plants on the microbial quality of the river. This indicates that treated wastewater effluents discharged from these treatment plants are a source of contamination of receiving surface waters with this potential pathogen. Upstream of the river at the NGWTP is an informal settlement with poor sanitation and inadequate sewage disposal system, which contaminates the river with human and animal wastes while the bank of the Umgeni River downstream, is littered with feces. Storm runoff from this informal settlement and riverbanks explains the high count of presumptive *Salmonella* spp. observed upstream. In contrast to findings from this study, Olaniran, Naidoo and Pillay \[[@B24-ijerph-12-09692]\] reported lower counts of *Salmonella* spp. from treated wastewater of same plants under investigation, which may be due to the short duration of the study or an improvement in the operational conditions of the treatment plants. Elsewhere, Momba, Osode and Sibewu \[[@B6-ijerph-12-09692]\] reported recovery of pathogenic microorganisms including *Salmonella* spp. in the final effluents of four WWTPs in the Eastern Cape province of South Africa and concluded that WWTPs serve as a point source of microbial pollution of water bodies. Recent reports have also suggested that WWTPs plants in South Africa are either dysfunctional or non-functional \[[@B41-ijerph-12-09692]\], inefficient in removing microbial pathogens from wastewater and produce wastewater effluent of unacceptable standard that does not meet discharge guidelines set by the department of water affairs, South Africa \[[@B42-ijerph-12-09692],[@B43-ijerph-12-09692],[@B44-ijerph-12-09692],[@B45-ijerph-12-09692]\]. The issue of treatment efficiency is of major importance if the reclaimed water is intended for recreational or potable reuse or is to be discharged into water bodies because disposal of inadequately treated wastewater into surface water recipient is one of the major sources of pathogens in the environment \[[@B42-ijerph-12-09692],[@B46-ijerph-12-09692]\]. Though *Salmonella* is isolated from water in lower numbers than indicator bacteria such as fecal coliform, fecal streptococci and enterococci; counts in the range of 15−1000 cfu/mL may pose public health risks \[[@B47-ijerph-12-09692]\]. Thus, the presence of this organism in the final effluent of wastewater and receiving surface water is a serious cause for concern especially where the contaminated water is depended on for irrigation and rural socio-economic activities. In this study, 200 *Salmonella* spp. were recovered and found to contain the *inv*A gene ([Figure 2](#ijerph-12-09692-f002){ref-type="fig"}), which led to their confirmation as *Salmonella* spp. in agreement with previous studies \[[@B48-ijerph-12-09692],[@B49-ijerph-12-09692]\]. The *inv*A gene is conserved in all *Salmonella* spp. and encodes for a protein in the inner and outer membrane which is essential for virulence and is thought to trigger the internalization required for invasion into deeper tissues \[[@B29-ijerph-12-09692],[@B30-ijerph-12-09692]\]. At the NWWTW, confirmed *Salmonella* spp. were only recovered at the B.C point (6.5%) but not at the D.P indicating the plant was efficient at removing *Salmonella* spp. from the wastewater during the sampling period. This is also corroborated by the fact that the NWWTW has a high Greendrop status rating \[[@B50-ijerph-12-09692]\]. The Greendrop is an index developed by the Department of Water Affairs in South Africa to rate wastewater treatment plants in the country based on the quality of their final effluent. In the Eastern Cape province of South Africa, Momba, Osode and Sibewu \[[@B6-ijerph-12-09692]\] observed the presence of *Salmonella* spp. in 50% of final wastewater effluent and 35% in the receiving river samples. Poor sanitation, lack of access to proper sewage disposal systems, malnutrition and poverty have been described as some of the leading factors contributing to the high prevalence of salmonellosis and other diarrheal diseases in developing countries \[[@B51-ijerph-12-09692],[@B52-ijerph-12-09692],[@B53-ijerph-12-09692]\]. Amplification of virulence genes ([Figure 2](#ijerph-12-09692-f002){ref-type="fig"}) revealed that the isolates contained one or more virulence genes present in the *Salmonella* pathogenicity island (SPI). This can pose serious health threats to consumers who depend on the river water for daily and recreational activities. Pathogenicity islands are found on genomes of pathogenic bacteria but are absent in non-pathogenic strains of the same or related species \[[@B54-ijerph-12-09692]\]. The presence of all four virulence genes were reported to be present in 87.2% of *Salmonella* isolated from patients with systemic infection \[[@B55-ijerph-12-09692]\] while 12.8% of *Salmonella* spp. isolated from stool samples lacked the *mis*L and *orf*L gene. The *spi*C gene is found in the SPI-2 and is essential for systemic pathogenesis because it encodes a type III secretion system (T3SS) that is activated after invasion \[[@B56-ijerph-12-09692],[@B57-ijerph-12-09692]\]. The T3SS system is used by the pathogen to deliver virulence factors to the host cell and interfere with or subvert normal host cell signaling pathways \[[@B58-ijerph-12-09692]\]. The *mis*L gene located on SPI-3 encodes an autotransporter protein involved in intestinal colonization and essential for survival in macrophages \[[@B55-ijerph-12-09692],[@B59-ijerph-12-09692]\]. The SPI-4 is a 25 kb pathogenicity island containing the *orf*L gene thought to encode a type 1 secretion system (an autotransporter protein) that mediate the secretion of toxins and is necessary for macrophage survival \[[@B55-ijerph-12-09692],[@B59-ijerph-12-09692]\], while the *pip*D gene encodes effector proteins for the T3SS transport protein \[[@B29-ijerph-12-09692],[@B30-ijerph-12-09692]\] and is mainly associated with enteropathogenesis \[[@B58-ijerph-12-09692]\]. The presence of these virulence genes in *Salmonella* spp. isolated from treated wastewater effluent and receiving surface water indicate the capabilities of these isolates in causing infections in susceptible hosts. Recently, there was report of an outbreak of acute gastroenteritis in KwaZulu-Natal, which was linked to food, contaminated with *Salmonella enterica serovar Enteritidis* resulting in the hospitalization of 216 people \[[@B60-ijerph-12-09692]\]. The report suggested a point source outbreak with a possibility of continued transmission. The true burden of *Salmonella* disease in Africa is unclear thus a comprehensive epidemiological study is needed to elucidate it. The *Salmonella* serotypes that most commonly cause invasive non-typhoidal *Salmonella* in Africa are *S. typhimurium* and *S. enteritidis*, which are usually associated with a broad host range and with enteric diseases \[[@B11-ijerph-12-09692]\]. Resistance against sulfamethoxazole (100%), nalidixic acid (27%) and streptomycin (14%) was observed among these isolates. Intermediate resistance was observed against streptomycin (74%), nalidixic acid (44%) and fosfomycin (8.5%). Resistance to nalidixic acid suggests possible resistance to third generation quinolones CLSI \[[@B31-ijerph-12-09692]\]. However, this was not observed in this study, as isolates resistant to nalidixic acid were also susceptible to the third generation quinolones tested. Previous studies have suggested that quinolones should not be used in the treatment of invasive Salmonellosis due to strains with decreased sensitivity to fluoroquinolones and possible risk of treatment failure \[[@B29-ijerph-12-09692],[@B61-ijerph-12-09692]\]. The results obtained in this study further emphasize the need for prudent use of fluoroquinolones and other commonly used antibiotics to prevent the emergence of resistant phenotypes \[[@B62-ijerph-12-09692]\]. Consistent with this study, *Salmonella* spp. were reported to be highly sensitive to third generation β-lactams \[[@B29-ijerph-12-09692],[@B59-ijerph-12-09692]\] but resistant to sulfamethoxazole, nalidixic acid and streptomycin. Data from the National Antimicrobial Resistance Monitoring Systems (NARMS) in the US from 1996 to 2004 showed increase in resistance of clinical isolates of *Salmonella* against antibiotics \[[@B63-ijerph-12-09692]\]. The upsurge in multidrug resistant strains of *Salmonella* over the past decade is threatening successful treatment of diseases caused by this organism especially in developing countries where disease burden is high \[[@B64-ijerph-12-09692]\]. In conclusion, this study revealed that treated effluents from WWTPs investigated are reservoirs of antibiotic resistant and virulent *Salmonella* spp. The isolates were susceptible to most third generation β lactams tested whilst exhibiting resistance to other antibiotics. The presence of virulence genes is indicative of their capabilities to cause infection in susceptible hosts. Though certain physicochemical parameters were within the stipulated guidelines, the overall quality of the treated effluent is still low. Thus, in order to protect the valuable surface water resources, urgent intervention is required by the regulatory authorities and workers in these treatment plants to optimize treatment efficiency. Constant surveillance of the treatment processes and final effluent, infrastructural upgrade of the wastewater treatment works and provision of adequate sanitation and sewage disposal systems to rural communities on the banks of rivers is recommended. This study was supported by the Medical Research Council and the National Research Foundation of South Africa. Financial support from the College of Agriculture, Engineering and Science, University of KwaZulu-Natal in the form of a bursary to the first author is also acknowledged. Ejovwokoghene C. Odjadjare was involved in the acquisition and analysis of data, statistical analysis and manuscript draft. Ademola O. Olaniran conceptualized and designed the research, was involved in the research coordination, interpretation of data and critically reviewed the draft manuscript. Both authors read and approved the final manuscript. The authors declare no conflict of interest.