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Is peanut-lupine antibody cross-reactivity associated to cross-allergenicity in peanut-sensitized mouse strains?

Peanut hypersensitivity is one of the most common food allergies and one of the most common causes of death by food anaphylaxis in children and adults. Cross-reactivity of peanut-specific antibody (Ab) with other legumes is frequently demonstrated but it still remains to be demonstrated whether these responses could lead to clinical signs of cross-allergenicity. We sought to evaluate peanut-specific serum IgE and IgG1 antibody (Ab) responses and anaphylactic reaction in mice strains and to assess both cross-reactivity and cross-allergenicity of peanut and lupine. Four mice strains (i.e., C3H, BALB/c, CBA and SJL) were sensitized to peanut by intraperitoneal (ip) injection of crude peanut protein extract with alum. Other groups were given oral peanut extract without adjuvant. Peanut-specific antibodies (Abs) and anaphylactic responses to peanut challenge were examined. The C3H, CBA (H-2(k)) and BALB/c (H2-(d)) mice exhibited high levels of peanut-specific serum IgE, IgG1 Ab responses after the intra-peritoneal sensitization. Only the two strains of mice in the H-2(k) background developed anaphylactic symptoms upon intra-peritoneal challenge with crude peanut protein extract. While cross-reactivity of peanut and lupine was confirmed by ELISA, no clinical symptom of cross-allergenicity was seen after challenge with lupine. Mice that were given oral peanut showed only increase in peanut-specific IgG2a, but no IgE or IgG1 Abs and failed to develop anaphylactic reactions following injection of either peanut or lupine protein.

These results show that mice of different genetic backgrounds can be sensitized to peanut by ip injection to develop anti-peanut Abs that cross react with lupine. In addition, cross-allergenicity may not directly correlate with the presence of cross-reactive Abs since no clinical symptoms of cross-allergenicity was seen after ip challenge with lupine.

Does atorvastatin attenuate sympathetic hyperinnervation together with the augmentation of M2 macrophages in rats postmyocardial infarction?

Inflammation after myocardial infarction (MI) causes cardiac nerve sprouting and consequent ventricular arrhythmias (VAs). Macrophages, as major immune cells, are involved in the entire inflammation response process and serve as a link between inflammation and sympathetic hyperinnervation by regulating nerve growth factor (NGF) expression. Accumulating evidence shows that statins possess antiarrhythmogenic properties, and the aim of this study was to explore the mechanism by which atorvastatin ameliorates cardiac sympathetic nerve sprouting via regulating macrophage polarization. Rat models of MI were created by ligating the left coronary artery. MI-operated rats received either atorvastatin or phosphate-buffered saline for 7 days. Immunohistochemical analyses and immunofluorescence staining were used to analyze macrophage infiltration after MI and to detect the distribution and density of growth-associated protein-43 (GAP-43) and tyrosine hydroxylase (TH) in nerve fibers in peri-infarct zones after MI. The polarity of the macrophages that were obtained from the rat peritoneal cavity was examined via flow cytometry. The protein levels of NGF were detected via Western blot analysis, and the concentrations of NGF in the supernatants were determined via enzyme-linked immunosorbent assay. The mRNA levels of NGF, inducible nitric oxide synthase (iNOS), Arginase-1 (Arg1), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) were examined by quantitative real-time polymerase chain reaction. The association between VAs and MI was evaluated using programmed electrophysiological stimulation. Macrophages were successfully induced to the M1 (classically activated) and M2 (alternatively activated) phenotypes by stimulation with lipopolysaccharide and interferon-γ (LPS + IFN-γ) and interleukin-4 (IL-4), respectively. Atorvastatin markedly downregulated IL-1β, TNF-α, iNOS, and NGF and upregulated Arg1, shifting the macrophage phenotype from M1 to M2. Moreover, atorvastatin significantly reduced TH levels and the density of GAP-43-positive nerve fibers and decreased inducible VAs.

Atorvastatin effectively ameliorated cardiac sympathetic nerve remodeling and prevented VAs after MI by significantly downregulating the expression of NGF, IL-1β, and TNF-α via together with the augmentation of M2 macrophage.

Does a prostacyclin analog beraprost sodium attenuate radiocontrast media-induced LLC-PK1 cells injury?

We previously reported that the apoptotic injury in a porcine renal tubular cell line LLC-PK1 cells induced by radiographic contrast media is attenuated by dibutyl cyclic adenosine monophosphate (cAMP) in a manner dependent on protein kinase A (PKA). The present study was designed to determine whether the elevation of endogenous cAMP with beraprost sodium, a prostacyclin analog, reduces the contrast material-induced renal tubular injury. The cell injury was induced by the exposure to ioversol for 30 minutes followed by further incubation for 24 hours in the absence of the contrast medium, and assessed by propidium iodide uptake and WST-8 assay. Apoptosis was determined by annexin V stain and DNA electrophoresis. Caspase activity was assessed by the enzymatic degradation of specific substrate peptides. Bax and bcl-2 mRNA expression were determined by reverse transcription-polymerase chain reaction (RT-PCR). The phosphorylation of cAMP-responsive element binding protein (CREB) was measured by an immunofluorescent method. Beraprost sodium (10 to 1000 nmol/L) attenuated concentration dependently the ioversol-induced decrease in cell viability, in which the protective effect of beraprost sodium was dependent on the elevation of cellular cAMP content. The phosphorylation of CREB was enhanced by beraprost sodium in PKA-dependent manner. In addition, beraprost sodium reversed the ioversol-induced increase in bax mRNA with a concomitant decrease in bcl-2 mRNA and subsequent activation of caspase-3 and -9, thereby resulting in the inhibition of the nuclear damage.

Beraprost sodium reversed the contrast media-induced renal tubular cells in culture by activating cAMP/protein kinase A-dependent phosphorylation of CREB and subsequent enhancement of bcl-2 expression.

Does supercritical CO2 decaffeination of unroasted coffee beans produce melanoidins with distinct NF-κB inhibitory activity?

The supercritical CO(2)-decaffeination process causes unroasted coffee beans to turn brown. Therefore, we suspected that the decaffeinated beans contained melanoidins. Decaffeinated unroasted coffee extract absorbed light at 405 nm with a specific extinction coefficient, K(mix 405 nm), of 0.02. Membrane dialysis (molecular weight cut-off, 12 to 14 kDa) increased the K(mix 405 nm) value 15 fold. Gel filtration chromatography showed that the high-MW fraction (MW > 12 kDa) had an elution profile closer to that of melanoidins of medium-roast coffee than to the corresponding fraction of unroasted coffee, indicating the presence of melanoidins in decaffeinated unroasted beans. Using murine myoblast C2C12 cells with a stably transfected nuclear factor-κB (NF-κB) luciferase reporter gene, we found that the high-MW fraction of decaffeinated unroasted beans had an NF-κB inhibitory activity of IC(50) = 499 μg/mL, more potent than that of regular-roast coffee (IC(50) = 766 μg/mL). Our results indicate that melanoidins form during the supercritical CO(2)-decaffeination process and possess biological properties distinct from those formed during the regular roasting process.

We discovered the roasting effect of decaffeination process, reporting the discovery of melanoidins in green (unroasted) decaf coffee beans. Our results indicated that melanoidins form during the supercritical CO2-decaffeination process and possess biological properties distinct from those formed during the regular roasting process. Our results offer new insights into the formation of bioactive coffee components during coffee decaffeination process.

Does cell-to-cell contact induce mesenchymal stem cell to differentiate into cardiomyocyte and smooth muscle cell?

Recent evidences have suggested that stem cell can differentiate into cardiomyocyte and smooth muscle cell (SMC) in vivo or in vitro. But the mechanism on how stem cell differentiates is still unknown. We investigated whether intercellular interaction or soluble chemical factors would induce mesenchymal stem cells (MSCs) to acquire the phenotypical characteristics of cardiomyocytes or SMC. MSCs were isolated from rat bone marrow with density gradient centrifugation and amplified in vitro. Flow cytometry was used to monitor the expression of surface antigen profile. After labeled by GFP (green fluorescent protein) transfection, rat MSCs were used to culture with adult rat cardiomyocytes and rat aortic SMCs in direct co-culture, indirect co-culture and conditioned culture, respectively. One week later, immunofluorescence staining against alpha-actin, desmin, and cardiac troponin T (cTnT) for cardiomyocyte, smooth muscle calponin and SM-alpha-actin for SMC were performed. Immunofluorescence staining was positive against alpha-actin, desmin, and cTnT on MSCs in co-culture group with adult cardiomyocytes, positive against smooth muscle calponin and SM-alpha-actin on MSCs in co-culture group with SMCs. In contrast, no alpha-actin, desmin, and cTnT expression was observed in the indirect co-culture group and conditioned culture group; no smooth muscle calponin and SM-alpha-actin in the indirect co-culture group and conditioned culture group.

Direct cell-to-cell contact between MSC and adult cardiomyocyte or SMC, but not the soluble signaling molecules is obligatory in the differentiation of MSC into cardiomyocytes or SMC.

Is ala/Thr60 variant of the Leydig insulin-like hormone associated with cryptorchidism in the Japanese population?

Leydig insulin-like hormone (Insl3), a member of the insulin-like superfamily, is specifically expressed in Leydig cells of fetal and postnatal murine testis. Recently, the absence of the Insl3 gene has been reported to result in bilateral cryptorchidism in male mice and it has been suggested that mutations of the INSL3 gene may cause cryptorchidism in humans. We sequenced the INSL3 gene from five Japanese patients with sporadic bilateral cryptorchidism. Patients' genome DNA was prepared from blood leukocytes. Two exons of the INSL3 gene were amplified by polymerase chain reaction and were sequenced directly. A restriction fragment length polymorphism assay was performed on 70 control samples for analysis of polymorphism. Three of five cases had a heterozygous single-base change, a G to A transition at position 178 of the INSL3 gene, which predicts an alanine (GCC) to threonine (ACC) change at codon 60 (designated A60T). However, the A60T mutation was also found in the normal Japanese population at an allele frequency of 26%, which suggests that this mutation is a common polymorphism and is not associated with the occurrence of cryptorchidism.

No mutation has been found in the INSL3 gene from Japanese patients with idiopathic cryptorchidism. We did find the A60T polymorphism, which was not associated with the occurrence of cryptorchidism.

Does EuroSCORE II perform better than its original versions?

The European system for cardiac operation risk evaluation (EuroSCORE) is widely used for predicting in-hospital mortality after cardiac surgery. A new score (EuroSCORE II) has been recently developed to update the previously released versions. This study was undertaken to validate EuroSCORE II, to compare its performance with the original EuroSCOREs and to evaluate the effects of the removal of those factors that were included in the score even if they were statistically non-significant. Data on 12,325 consecutive patients who underwent major cardiac surgery in a 6-year period were retrieved from three prospective institutional databases. Discriminatory power was assessed using the c-index and comparison among the scores' performances was performed with Delong, bootstrap, and Venkatraman methods. Calibration was evaluated with calibration curves and associated statistics. The in-hospital mortality rate was 2.2%. The discriminatory power was high and similar in all algorithms (area under the curve 0.82, 95% CI: 0.79-0.84 for additive EuroSCORE; 0.82, 95% CI: 0.79-0.84 for logistic EuroSCORE; 0.82, 95% CI: 0.80-0.85 for EuroSCORE II). The EuroSCORE II had a fair calibration till 30%-predicted values and over-predicted beyond. The removal of non-significant factors from EuroSCORE II did not affect performance, being both the calibration and discrimination comparable.

This validation study demonstrated that EuroSCORE II is a good predictor of perioperative mortality. It showed an optimal calibration until 30%-predicted mortality. Nonetheless, it does not seem to significantly improve the performance of older versions in the higher tertiles of risk. Moreover, it could be simplified, as the removal from the algorithm of non-significant factors does not alter its performance.

Does nicotine metabolite ratio predict efficacy of transdermal nicotine for smoking cessation?

Nicotine is metabolized to cotinine, and cotinine is metabolized to 3'-hydroxycotinine (3-HC) by the liver enzyme cytochrome P450 (CYP) 2A6. More rapid metabolism of nicotine may result in lower nicotine blood levels from nicotine replacement products and poorer smoking cessation outcomes. This study evaluated the utility of the 3-HC/cotinine ratio as a predictor of the efficacy of nicotine replacement therapy as an aid for smoking cessation. By use of an open-label design, 480 treatment-seeking smokers were randomly assigned to 8 weeks of transdermal nicotine or nicotine nasal spray use, plus behavioral group counseling. Assessments included demographics, smoking history, body mass index, and plasma nicotine, cotinine, and 3-HC concentrations, as well as CYP2A6 genotypes. Smoking cessation was biochemically verified at the end of treatment and at 6-month follow-up. The rate of nicotine metabolism, as indicated by pretreatment 3-HC/cotinine ratio derived from cigarette smoking, predicted the effectiveness of transdermal nicotine at both time points. The odds of abstinence were reduced by almost 30% with each increasing quartile of metabolite ratio (odds ratio, 0.72 [95% confidence interval, 0.57-0.90]; P=.005). Higher metabolite ratios also predicted lower nicotine concentrations (beta=-1.72, t(179)=-3.31, P<.001), as well as more severe cravings for cigarettes after 1 week of treatment (beta=0.32, t(190)=2.91, P=.004). The metabolite ratio did not predict cessation with use of nicotine nasal spray (odds ratio, 1.05 [95% confidence interval, 0.83-1.33]; P=.68).

The nicotine metabolite ratio might be useful in screening smokers to determine likely success with a standard dose of transdermal nicotine.

Do genetic variations of IL17F and IL23A show associations with Behçet 's disease and Vogt-Koyanagi-Harada syndrome?

To investigate the associations of IL17A, IL17F, IL23A, and IL23R copy number variants (CNVs) with Vogt-Koyanagi-Harada (VKH) syndrome and Behçet's disease (BD) and the possible mechanisms involved. Two-stage case-control and functional studies. A total of 1159 VKH patients, 1036 BD patients, and 2050 controls were enrolled. TaqMan real-time polymerase chain reaction assay was used for genotyping of copy number variant. Cell proliferation was measured by colorimetric assay. Association of CNVs in IL17A, IL17F, IL23A, and IL23R with BD and VKH syndrome and the functional roles of IL17F CNVs. Increased frequencies of more than 2 copies of IL17F and IL23A were found in BD patients as compared with controls (IL17F: P=4.17×10(-8); odds ratio [OR], 2.2; IL23A: P=2.86×10(-11); OR, 2.8, respectively). A similar result was found for VKH syndrome (IL17F: P=2.84×10(-13); OR, 2.7; IL23A: P=4.46×10(-17); OR, 3.4, respectively). Interestingly, the association of IL17F and IL23A with BD was found only in male patients (IL17F: P=1.06×10(-6); OR, 2.3; IL23A, P=3.81×10(-8); OR, 2.8, respectively), but not in female patients. No association of CNVs in IL17A and IL23R was found for BD and VKH syndrome. IL17F protein levels were correlated positively with gene copy numbers (P=3.43×10(-7)). Individuals with high IL17F copies showed enhanced peripheral blood mononuclear cells (PBMC) proliferation (P=5.67×10(-3)).

High gene copy numbers of IL17F and IL23A were associated with BD and VKH syndrome. Enhanced IL17F protein production and PBMC proliferation were associated with high IL17F copy numbers.

Is minimally invasive staging of endometrial cancer feasible and safe in elderly women?

To compare the surgical outcome of elderly and younger patients undergoing laparoscopic or robotic surgical staging of endometrial cancer. Retrospective analysis (Canadian Task Force classification II-2). University-affiliated hospital. One hundred twenty-nine patients comprised the study group. Sixty patients were aged 65 years or older (elderly group), and 69 patients were younger than 65 years (younger group). Abdominal, laparoscopic, or robotic hysterectomy. Among the 109 patients who underwent laparoscopic or robotic staging, there were no differences in estimated blood loss, lymph node count, surgical time, complications, rate of blood transfusion, conversion to laparotomy, and mean postoperative stay between elderly and younger patients.

Minimally invasive surgical staging for endometrial cancer is both feasible and safe in the elderly population and offers similar outcomes as in younger patients.

Are exercise-induced abnormal blood pressure responses related to subendocardial ischemia in hypertrophic cardiomyopathy?

We examined by thallium-201 scintigraphy whether exercise-induced abnormal blood pressure response (BPR) is related to myocardial ischemia. Hemodynamic instabilities during exercise in patients with hypertrophic cardiomyopathy (HCM) are considered to be caused by abnormal reflex control of vascular resistance. In 105 patients with HCM, exercise thallium scintigraphy was performed by means of a multistage, symptom-limited bicycle ergometer exercise test. Eighty-eight patients had normal BPR (> or = 25 mm Hg from baseline to peak exercise), and 17 had abnormal BPR (<25 mm Hg). Clinical characteristics including age, the prevalence of obstruction, New York Heart Association functional class and echocardiographic measurements were similar between the two groups. Left ventricular end-diastolic pressure was significantly higher in patients with abnormal BPR than in those with normal BPR (18+/-8 vs. 14+/-5 mm Hg, p < 0.05). Exercise-induced perfusion abnormalities including fixed and reversible perfusion defects, and left ventricular cavity dilatation (LVCD) were identified in 72 (69%) of 105 study patients. Left ventricular cavity dilatation indicates subendocardial hypoperfusion and is a marker of diffuse subendocardial ischemia. The prevalence of fixed or reversible perfusion defects was similar between the two groups. Patients with abnormal BPR had the higher prevalence of LVCD as compared to those with normal BPR (47.1 vs. 10%, p < 0.0002). Multiple logistic regression analysis revealed that LVCD was independently associated with abnormal BPR (odds ratio 3.76, 95% confidence interval 1.61 to 8.76).

Exercise-induced abnormal BPRs in patients with HCM are related to subendocardial ischemia during exercise.

Is free to total prostate-specific antigen ( PSA ) ratio superior to total-PSA in differentiating benign prostate hypertrophy from prostate cancer?

Serum prostate-specific antigen (PSA) exists in different molecular forms, and their respective concentration has been proposed as a useful tool to improve discrimination between benign prostatic hypertrophy (BPH) and prostate cancer (PC). The relevance of the free to total PSA ratio was prospectively studied in a selected urology clinic population of 420 patients. Total serum PSA ranged from 2.1 to 30 ng/ml; 154 had PC and 266 had BPH. Receiver operating characteristic (ROC) curves were constructed for the total population (total-PSA range from 2.1 to 30 ng/ml) and for the diagnostic gray zone of 2.1-10 ng/ml. For the two groups, the free to total PSA ratio had a higher specificity than total-PSA for all sensitivity levels. Cut-off values were found to, vary with prostate weight.

Although free to total PSA ratio demonstrated better performances than total-PSA, its use in screening appears problematic, due to the low prevalence of prostate cancer.

Do severe postoperative complications adversely affect long-term survival after R1 resection for pancreatic head adenocarcinoma?

Survival after pancreatic head adenocarcinoma surgery is determined by tumor characteristics, resection margins, and adjuvant chemotherapy. Few studies have analyzed the long-term impact of postoperative morbidity. The aim of the present study was to assess the impact of postoperative complications on long-term survival after pancreaticoduodenectomy for cancer. Of 294 consecutive pancreatectomies performed between January 2000 and July 2011, a total of 101 pancreatic head resections for pancreatic ductal adenocarcinoma were retrospectively analyzed. Postoperative complications were classified on a five-grade validated scale and were correlated with long-term survival. Grade IIIb to IVb complications were defined as severe. Postoperative mortality and morbidity were 5 and 57 %, respectively. Severe postoperative complications occurred in 16 patients (16 %). Median overall survival was 1.4 years. Significant prognostic factors of survival were the N-stage of the tumor (median survival 3.4 years for N0 vs. 1.3 years for N1, p = 0.018) and R status of the resection (median survival 1.6 years for R0 vs. 1.2 years for R1, p = 0.038). Median survival after severe postoperative complications was decreased from 1.9 to 1.2 years (p = 0.06). Median survival for N0 or N1 tumor or after R0 resection was not influenced by the occurrence and severity of complications, but patients with a R1 resection and severe complications showed a worsened median survival of 0.6 vs. 2.0 years without severe complications (p = 0.0005).

Postoperative severe morbidity per se had no impact on long-term survival except in patients with R1 tumor resection. These results suggest that severe complications after R1 resection predict poor outcome.

Is dehydration the most common indication for readmission after diverting ileostomy creation?

Early readmission after discharge from the hospital is an undesirable outcome. Ileostomies are commonly used to prevent symptomatic anastomotic complications in colorectal resections. The aim of this study was to identify factors predictive of readmission after colectomy/proctectomy and diverting loop ileostomy. This study is a retrospective review. Patients were included who underwent colon and rectal resections with ileostomy at our institution. Sex, age, type of disease, comorbidities, elective vs urgent procedure, type of ileostomy, operative method, steroid use, ASA score, and the use of diuretics were evaluated as potential factors for readmission. The primary outcomes measured were the need for readmission and the presence of dehydration (ostomy output ≥1500 mL over 24 hours and a blood urea nitrogen/creatinine level ≥20, or physical findings of dehydration). Six hundred three loop ileostomies were created mostly in white (95.3%), male (55.6%) patients undergoing colon or rectal resections. IBD was the most common indication at 50.9%, with rectal cancer at 16.1%, and other at 31.0%. The 60-day readmission rate was 16.9% (n = 102) with the most common cause dehydration (n = 44, 43.1%). Regression analysis demonstrated that the laparoscopic approach (p = 0.02), lack of epidural anesthesia (p = 0.004), preoperative use of steroids (p = 0.04), and postoperative use of diuretics (p = 0.0001) were highly predictive for readmission. Furthermore, regression analysis for readmission for dehydration identified the use of postoperative diuretics as the sole risk factor (p = 0.0001).

This study is limited by the retrospective analysis of data, and it does not capture patients that were treated at home or in clinic.

Does peripheral neuropathy induce cutaneous hypersensitivity in chronically spinalized rats?

The present study was aimed at the issue of whether peripheral nerve injury-induced chronic pain is maintained by supraspinal structures governing descending facilitation to the spinal dorsal horn, or whether altered peripheral nociceptive mechanisms sustain central hyperexcitability and, in turn, neuropathic pain. We examined this question by determining the contribution of peripheral/spinal mechanisms, isolated from supraspinal influence(s), in cutaneous hypersensitivity in an animal model of peripheral neuropathy. Adult rats were spinalized at T8-T9; 8 days later, peripheral neuropathy was induced by implanting a 2-mm polyethylene cuff around the left sciatic nerve. Hind paw withdrawal responses to mechanical or thermal plantar stimulation were evaluated using von Frey filaments or a heat lamp, respectively. Spinalized rats without cuff implantation exhibited a moderate decrease in mechanical withdrawal threshold on ~day 10 (P < 0.05) and in thermal withdrawal threshold on ~day 18 (P < 0.05). However, cuff-implanted spinalized rats developed a more rapid and significant decrease in mechanical (~day 4; P < 0.001) and thermal (~day 10; P < 0.05) withdrawal thresholds that remained significantly decreased through the duration of the study.

Our findings demonstrate an aberrant peripheral/spinal mechanism that induces and maintains thermal and to a greater degree tactile cutaneous hypersensitivity in the cuff model of neuropathic pain, and raise the prospect that altered peripheral/spinal nociceptive mechanisms in humans with peripheral neuropathy may have a pathologically relevant role in both inducing and sustaining neuropathic pain.

External benchmarking of trauma center performance: have we forgotten our elders?

The elderly injured have been identified as a population with unique needs compared with nonelderly trauma patients. We sought to determine whether trauma center (TC) performance is consistent across age groups and to assess whether aggregate evaluations of TC performance capture quality of care among the elderly. The recently launched Trauma Quality Improvement Program utilizes external benchmarking of TC outcomes to identify centers with above-average performance, with the goal of disseminating best practices. If variation exists in TC performance across age groups, such variation might significantly impact on the success of external benchmarking programs in improving quality of care. Study data were derived from the National Trauma Databank (2007), limited to level I and II centers and adults with moderate to severe injuries (injury severity score>9). Separate logistic regression models were constructed to produce TC risk-adjusted mortality for both the young and the elderly (age>65 years). Observed-to-expected mortality ratios were used to identify centers with above or below average performance overall, among the young and among the elderly. We identified 87,754 patients across 132 facilities; 25% were elderly. After adjustment for case mix, 9 centers were identified as above-average performers in the elderly population. Only 2 of these centers were also above-average performers among young patients. Overall, concordance for center performance across age strata evidenced poor agreement (κ, 0.23). In addition, aggregate assessment of center performance did not reliably identify high-performing centers for elderly patients.

The use of outcome-based benchmarking harbors significant potential for trauma quality improvement. Evaluations of aggregate TC performance may not adequately reflect the care provided to the elderly injured. Elderly trauma patients may warrant special attention in the context of ongoing quality improvement programs.

Does l-3-n-butylphthalide promote Neurogenesis and Neuroplasticity in Cerebral Ischemic Rats?

This study investigated whether anticerebral ischemia new drug, l-3-n-butylphthalide (l-NBP), improved behavioral recovery and enhanced hippocampal neurogenesis after cerebral ischemia in rats. The middle cerebral artery of rats was blocked for 2 h. The daily oral administrations of 30 mg/kg l-NBP or vehicle were begun from the second day until the rats were sacrificed. L-NBP treatment markedly increased 5-bromo-2'-deoxyuridine (BrdU)-positive cells in the hippocampal dentate gyrus (DG) of injured hemisphere on day 28 after ischemia. The amount of newborn cells and newly mature neurons was also increased. The expressions of growth-associated protein-43 and synaptophysin were significantly elevated in l-NBP-treated rats. However, l-NBP markedly reduced the percentage of BrdU(+) /GFAP(+) cells. Additionally, the levels of catalytical subunit of protein kinase A (PKA), protein kinase B (Akt), and cAMP response element-binding protein (CREB) were significantly increased, and the activation of the signal transducer and activation of transcription 3 (STAT3) and the expressions of cleaved caspase-3 and Bax were obviously inhibited by l-NBP. Consequently, l-NBP attenuated the behavioral dysfunction.

It first demonstrates that l-NBP may improve the behavioral outcome of cerebral ischemia by promoting neurogenesis and neuroplasticity. Activation of CREB and Akt and inhibition of STAT3 signaling might be involved in.

Do an allometric model to estimate fluid requirements in children following burn injury?

To evaluate the ability of an allometric 3/4 Power Model combined with the Galveston Formula (Galveston-3/4 PM Formula) to predict fluid resuscitation requirements in children suffering burn injuries in comparison with the frequently used Parkland Formula and Galveston Formula using the Du Bois formula for surface area estimation (Galveston-DB Formula). To demonstrate that the Galveston-3/4 PM Formula is clinically equivalent to the Galveston-DB Formula for the estimation of fluid requirements in pediatric burn injury cases. Fluid resuscitation requirements differ in children suffering burn injuries when compared to adults. The Parkland Formula works well for normal weight adults but underestimates fluid requirements when indiscriminately applied to pediatric burn patients. The Galveston-DB Formula accounts for the change in body composition with age by using a body surface area (BSA) model but requires the measurement of height. The allometric model, using an exponent of 3/4, accounts for the dependence of a physiological variable on body mass without requiring height measurement and can be applied to estimate fluid requirements after burn injury in children. Comparisons were performed between the hourly calculated fluid requirements for the first 8 h following 20%, 40%, and 60% BSA burns using the Parkland Formula, the Galveston-DB Formula and Galveston-3/4 PM Formula for children 2-23 kg. In children less than 23 kg, the fluid requirements predicted by the Galveston-3/4 PM Formula are well correlated with those predicted by the Galveston-DB Formula (R2 = 0.997, P < 0.0001) and are much better than of the predictions made with the Parkland Formula, especially for children <10 kg.

For the purposes of clinical estimation of fluid requirements, the Galveston-3/4 PM Formula is indistinguishable from the Galveston-DB Formula in children 23 kg or less.

Is amoxicillin/clavulanic acid ineffective at preventing otitis media in children with presumed viral upper respiratory infection : a randomized , double-blind equivalence , placebo-controlled trial?

To assess the equivalence of amoxicillin/clavulanic acid and placebo in the prevention of acute otitis media in children at high risk of acute otitis media who develop upper respiratory tract infection. This was a multicentre, equivalence, randomized, double-blind trial of two parallel groups comparing 5 days of amoxicillin/clavulanic acid 75 mg kg-1 day-1 (i.e. 25 mg kg-1 every 8 h) and placebo. The main outcome measure was acute otitis media occurring within 8-12 days of initiating treatment. Two hundred and three infants, aged 3 months-3 years with upper respiratory tract infection over 36 h and a history of recurrent acute otitis media were included over 8.5 months. Two children were lost to follow-up. Patient characteristics were similar in both groups. In the intention to treat analysis the frequency of acute otitis media was 16.2% (16/99) in the placebo group and 9.6% (10/104) in the amoxicillin/clavulanic acid group (P = 0.288). The difference between acute otitis media rates was 6.6% (one-sided 95% confidence interval of 14.3%). The occurrence of side-effects was similar in the amoxicillin/clavulanic acid and placebo groups.

The difference in effectiveness between antibiotic and placebo was not greater than 14.3%, and we calculated that 94 children would need to be exposed to antibiotics to avoid six cases of acute otitis media. In view of the risk of development of resistance due to frequent exposure to antibiotics, our study supports the need for reduction in the administration of antibiotics in upper respiratory tract infection even in children at high risk of acute otitis media.

Natural head position and lower incisor irregularity: Is there a relationship?

The aim of this study was to assess the relationship between dynamic measurements of natural head position (NHP) and lower incisor irregularity to identify potential gender differences. A total of 103 plaster models and dynamic NHP measurements were taken from 51 male (mean age: 14.20 ± 2.51 years) and 52 female (mean age: 15.02 ± 2.67 years) subjects. The dynamic NHP data were gathered by using an inclinometer device and a portable data logger. Lower incisor irregularity was measured with Little's irregularity index. The Mann-Whitney U and Kruskal-Wallis rank tests were used at a significance level of p<0.05. To evaluate the correlation between NHP and lower incisor irregularity, Spearman correlation coefficients (r) were calculated. There were significant gender differences in the sagittal measurements of NHP (p = 0.031) and incisor irregularity (p = 0.023). A Kruskal-Wallis test revealed no significant difference in NHP measurements between subjects presenting different levels of irregularity. Females displayed no significant correlation between incisor irregularity and any NHP measurement. However, in the males high correlation coefficients between incisor irregularity and sagittal NHP measurements (r = 0.369; p = 0.008) were noted.

Significant correlations between lower incisor irregularity and sagittal NHP measurements in males were observed. Females had a more forwardly inclined NHP than males. Moreover, male subjects displayed greater incisor irregularity than female subjects.

Do consistent relationships between sensory properties of savory snack foods and calories influence food intake in rats?

Determine the influence of experience with consistent or inconsistent relationships between the sensory properties of snack foods and their caloric consequences on the control of food intake or body weight in rats. Rats received plain and BBQ flavored potato chips as a dietary supplement, along with ad lib rat chow. For some rats the potato chips were a consistent source of high fat and high calories (regular potato chips). For other rats, the chips provided high fat and high calories on some occasions (regular potato chips) and provided no digestible fat and fewer calories at other times (light potato chips manufactured with a fat substitute). Thus, animals in the first group were given experiences that the sensory properties of potato chips were strong predictors of high calories, while animals in the second group were given experiences that the sensory properties of potato chips were not predictors of high calories. Juvenile and adult male Sprague-Dawley rats. Following exposure to varying potato chip-calorie contingencies, intake of a novel, high-fat snack food and subsequent chow intake were assessed. Body weight gain and body composition as measured by DEXA were also measured. In juvenile animals, exposure to a consistent relationship between potato chips and calories resulted in reduced chow intake, both when no chips were provided and following consumption of a novel high-fat, high-calorie snack chip. Long-term experience with these contingencies did not affect body weight gain or body composition in juveniles. In adult rats, exposure to an inconsistent relationship between potato chips and calories resulted in increased consumption of a novel high-fat, high-calorie snack chip premeal along with impaired compensation for the calories contained in the premeal.

Consumption of foods in which the sensory properties are poor predictors of caloric consequences may alter subsequent food intake.

Do children with benign rolandic epilepsy have a higher prevalence of migraine than those with other partial epilepsies or nonepilepsy controls?

Prior studies have given conflicting data concerning the association of benign rolandic epilepsy of childhood (BREC) and migraine but were limited by lack of sensitive, diagnostic criteria for childhood migraine. By using revised International Headache Society (IHS-R) criteria, we compared the prevalence of migraine in children with BREC with that of those (a) with cryptogenic/symptomatic partial epilepsy and (b) without epilepsy. Three cohorts of children, gender and age matched (within 1 year) were identified: (a) BREC, (b) cryptogenic/symptomatic partial epilepsy, and (c) no history of seizures. Parents were queried in a standardized interview about migraine and migraine equivalents in their child, and in either biologic parent. Migraine was defined by using the IHS-R (for children) and IHS criteria (for parents). Children with headache were divided into definite (meeting IHS-R criteria), probable (recurrent, throbbing headaches with nausea, vomiting, photophobia or phonophobia, not meeting IHS-R criteria), possible (recurrent headaches with throbbing character or associated nausea/vomiting), or nonmigraine groups. chi(2) analysis was used to determine whether the cohort with BREC had a higher prevalence of definite, definite or probable, or definite, probable, or possible migraines or migraine equivalents than the other two cohorts. Each cohort consisted of 53 children (mean age, 9.8-9.9 years, M/F ratio, 35:18). Those with BREC had higher rates of definite and probable (p = 0.05), of definite, probable, and possible migraine (p = 0.05), and of migraine equivalents excluding motion sickness (p<0.005) than did those without seizures; however, they did not differ significantly from the cryptogenic/symptomatic partial epilepsy cohort.

Partial epilepsy, regardless of etiology, is associated with higher rates of migraine in children. The pathophysiologic link between epilepsy and migraine is unknown.

Are bilateral weighted radiographs required for accurate classification of acromioclavicular separation : an observational study of 59 cases?

Misinterpretation of the Rockwood classification system for acromioclavicular joint (ACJ) separations has resulted in a trend towards using unilateral radiographs for grading. Further, the use of weighted views to 'unmask' a grade III injury has fallen out of favour. Recent evidence suggests that many radiographic grade III injuries represent only a partial injury to the stabilising ligaments. This study aimed to determine (1) whether accurate classification is possible on unilateral radiographs and (2) the efficacy of weighted bilateral radiographs in unmasking higher-grade injuries. Complete bilateral non-weighted and weighted sets of radiographs for patients presenting with an acromioclavicular separation over a 10-year period were analysed retrospectively, and they were graded I-VI according to Rockwood's criteria. Comparison was made between grading based on (1) a single antero-posterior (AP) view of the injured side, (2) bilateral non-weighted views and (3) bilateral weighted views. Radiographic measurements for cases that changed grade after weighted views were statistically compared to see if this could have been predicted beforehand. Fifty-nine sets of radiographs on 59 patients (48 male, mean age of 33 years) were included. Compared with unilateral radiographs, non-weighted bilateral comparison films resulted in a grade change for 44 patients (74.5%). Twenty-eight of 56 patients initially graded as I, II or III were upgraded to grade V and two of three initial grade V patients were downgraded to grade III. The addition of a weighted view further upgraded 10 patients to grade V. No grade II injury was changed to grade III and no injury of any severity was downgraded by a weighted view. Grade III injuries upgraded on weighted views had a significantly greater baseline median percentage coracoclavicular distance increase than those that were not upgraded (80.7% vs. 55.4%, p=0.015). However, no cut-off point for this value could be identified to predict an upgrade.

The accurate classification of ACJ separation requires weighted bilateral comparative views. Attempts to predict grade on a single AP radiograph result in a gross underestimation of severity. The value of bilateral weighted views is to 'unmask' a grade V injury, and it is recommended as a first-line investigation.

Investigation of often-reported ten percent hysteroscopy fluid overfill: is this accurate?

A key component of hysteroscopic complications, such as fluid overload and severe dilutional hyponatremia, is the failure to anticipate and quickly recognize fluid deficits. The purpose of this study was to measure the volume and mass of irrigation fluid bags to assess the overfill of 3 common types of hysteroscopy irrigation fluids, 0.9% normal saline solution, 3% sorbitol, and 1.5% glycine, to challenge the often-quoted standard of assumption that overfill may be as high as 10% of the bag's volume. Ten cases of irrigation fluid were tested. The volume and weight of drained fluid from 18 bags of 0.9% normal saline solution 2000 mL, 12 bags 3% sorbitol 3000 mL, 8 bags of 1.5% glycine 3000 mL, and 4 bags of 0.9% normal saline solution 5000 mL were measured. Institutional review board exemption was obtained. Ten cases of irrigation fluid were obtained. The volume and weight of drained fluid from 18 bags of 0.9% normal saline solution 2000 mL, 12 bags of 3% sorbitol 3000 mL, 8 bags of 1.5% glycine 3000 mL, and 4 bags of 0.9% normal saline solution 5000 mL were measured. By volume, varying by the type of fluid tested, the maximum observed overfill was between 3.3% to 5.0%. For confirmation, each bag was also weighed and found to have a maximum overfill between 2.8% to 5.6%, varying with the volume and type of fluid measured. These findings were then compared with the manufacturer-provided overfill range of 1.5% to 6.0%. No underfill was observed.

Contrary to assertions over the last 25 years that overfill is 10% or higher as a rule, it appears more reasonable to assume that the degree of overfill is contingent on the type and volume of fluid used and is more likely closer to 2.8% to 5.6%. Therefore an accurate collecting system and weight measurement is more precise.

Is the initial diagnostic impression of "noncardiac chest pain" adequate to exclude cardiac disease?

In patients presenting to the emergency department (ED) with an initial diagnostic impression of noncardiac chest pain, we determine the 30-day incidence of adverse cardiac events and characteristics associated with those events. The multicenter, prospectively collected Internet Tracking Registry for Acute Coronary Syndromes (i*tr ACS ) registry of patients with chest pain enrolled from June 1, 1999, to August 1, 2001, was reviewed. We included patients if the physician's initial diagnostic impression was noncardiac chest pain after the medical history, physical examination, and initial 12-lead ECG. ED records, inpatient records, and follow-up results were reviewed for evidence of an adverse cardiac event defined as ST-segment or non-ST-segment elevation myocardial infarction, unstable angina, revascularization, or cardiac death within 30 days. Of 17,737 patients enrolled in i*tr ACS , 2,992 had an initial emergency physician impression of noncardiac chest pain. Of these, 85 (2.8%) patients had definite evidence for an adverse cardiac event. The adverse cardiac event group was older (61.2 versus 47.9 years), more likely to be men (58.6% versus 38.7%), and had a higher Acute Cardiac Ischemia-Time Insensitive Predictive Instrument score (26.1 versus 15.6). Factors associated with adverse cardiac events included hypercholesterolemia, diabetes, history of coronary artery disease, and history of congestive heart failure.

When the initial impression is noncardiac chest pain, high-risk features such as traditional cardiovascular risk factors or a history of coronary artery disease are associated with adverse cardiac events. In the absence of well-defined criteria, treating physicians should consider further evaluation before diagnosing patients with noncardiac chest pain if these features are present.

Is participation in cardiac rehabilitation programs associated with better quality of life and return to work after coronary artery bypass operations?

To explore the putative effect of cardiac rehabilitation programs on the 'health-related quality of life' and 'return to work' in pre-retirement patients one year after coronary artery bypass grafting. Of the 2,085 patients aged 45-64 who survived one year after CABG and were Israeli residents, 145 (6.9%) had participated in rehabilitation programs. Of these, 124 (83%) who answered QOL questionnaires were individually matched with 248 controls by gender, age within 5 years, and the time the questionnaire was answered. All patients had full clinical follow-up including a pre-operative interview. The Short Form-36 QOL questionnaire as well as a specific questionnaire were mailed to surviving patients one year after surgery. Study outcomes included the scores on eight scales and two summary components of the SF-36, as well as 'return to work' and 'satisfaction with medical services' from the specific questionnaire. Analysis was done for matched samples. Cardiac rehabilitation participants had significantly higher SF-36 scores in general health, physical functioning, and social functioning. They had borderline significant higher scores in the physical summary component of the SF-36. The specific questionnaire revealed significantly better overall functioning, higher satisfaction with medical care, and higher rate of return to work. While participants in cardiac rehabilitation and their controls were similar in their socio-demographic and clinical profiles, participating patients tended to be more physically active and more fully employed than their controls.

Rehabilitation participants had a self-perception of better HRQOL, most significantly in social functioning. Our findings of more frequent return to work and higher satisfaction with medical care should induce a policy to encourage participation in cardiac rehabilitation programs after CABG.

Is social phobia related to lack of social skills?

The aim of the current study was to investigate whether social phobics differ from clinically anxious and non-clinical comparison groups with regard to either the duration of skill-related behaviours displayed during a social conversation or their behavioural adequacy as perceived by observers. A between-group design was used for the first part of the study. The duration of specified skill-related behaviours of a group of socially phobic participants was compared with the behaviours of clinically anxious and non-clinical participants. A within-group design was used for the second part of the study. Observers' ratings of the adequacy of the behaviour of a subgroup of the participants from the three groups were evaluated. Fifty-four participants, 18 socially phobic, 18 clinically anxious but not socially phobic and 18 who formed a non-clinical comparison group, took part in a 9-min conversation with a confederate of the experimenters. Participants were screened by diagnostic interview. Each conversation was videotaped, and from these recordings, the percentage time spent talking, spent in silence, smiling, maintaining eye contact while talking and while listening and while making manipulative gestures was extracted. A subset of 30 participants, 10 from each of the three groups, provided stimulus material for the second part of the study. The sixth minute of each of these 30 participants' recorded conversations was shown to 21 observers who rated the performance for adequacy of gestures, adequacy of eye contact, adequacy of smiling, clarity of speech, fluency of speech and overall adequacy of performance. Observers were also asked to categorize the participants based on the observed behaviour into one of three diagnostic groups: socially phobic, clinically anxious or non-clinical. Social phobics exhibited significantly less eye contact while talking in comparison with the non-clinical participants and significantly more manipulative gestures than either the clinically anxious or non-clinical participants. There were no other between-group differences in duration of skill-related behaviours. With regard to ratings of perceived behavioural adequacy for gestures, speech fluency and overall performance, the social phobic group was rated as significantly less adequate than both comparison groups. With regard to ratings of perceived adequacy of eye contact and speech clarity, while the anxious groups did not differ from each other, both were rated as significantly less adequate than the non-clinical comparison group. There was also a marked within-group variation in perceived adequacy of behaviour and wide variation in ratings provided by observers.

Possible competing explanations for the results are discussed; these include the skills deficit hypothesis and the possibility that group differences in both duration of skill-related behaviour and observed behavioural adequacy reflect awkward, anxious behaviours and/or safety behaviours.

Is mobilization of endothelial progenitor cells from bone marrow impaired in a piglet model of acute respiratory distress syndrome?

To characterize the endothelial progenitor cell mobilization in the models of moderate and severe lung injury, we hypothesized that there were differences in endothelial progenitor cell levels and mobilizing cytokines between moderate and severe lung injury. Prospective, randomized, and controlled experimental study. University research laboratory center. Fifteen healthy piglets. Piglets were randomly allocated to control, moderate lung injury (acute lung injury), and severe lung injury (acute respiratory distress syndrome) groups. Lung injury was established by intravenous infusion of oleic acid. Animals were mechanically ventilated for 24-48 hours, and then animals were weaned from ventilation and cared for until day 7. Endothelial progenitor cells were quantified by flow cytometry. After 24 hours, the number of endothelial progenitor cells in peripheral blood increased in the acute lung injury group but was not altered in the acute respiratory distress syndrome group compared to the control group. The number of CD34KDR, KDRCD133, and CD34KDRCD133 cells was higher in the acute lung injury group than in the acute respiratory distress syndrome group. In bone marrow, the number of CD34KDR and KDRCD133 cells was greater in acute respiratory distress syndrome animals but not altered in acute lung injury animals at 24 hours. Furthermore, plasma stromal cell-derived factor-1 and vascular endothelial growth factor concentrations were higher in acute lung injury than in acute respiratory distress syndrome at 24 hours. Matrix metalloproteinase-9 and soluble kit ligand levels in bone marrow were reduced in acute respiratory distress syndrome compared with acute lung injury. Lung CD34, KDR, and lung stromal cell-derived factor-1 messenger RNA expression were higher in the acute lung injury group than in the acute respiratory distress syndrome group. Furthermore, the expression of CD34, KDR, and CD133 messenger RNA in lung tissue was correlated with stromal cell-derived factor-1 in the lung.

There was a rapid release of endothelial progenitor cells from bone marrow into circulation in moderate acute lung injury, and endothelial progenitor cell mobilization was impaired in acute respiratory distress syndrome.

Is ineffective motility a marker for gastroesophageal reflux disease?

Previous studies have suggested that ineffective esophageal motility (IEM) may be a marker for gastroesophageal reflux disease (GERD), particularly supraesophageal reflux disease. We evaluated the relationship between esophageal acid exposure and esophageal body motility in patients undergoing both esophageal manometry and 24-h pH metry in the absence of antisecretory therapy. We conducted a retrospective database review of 84 patients (mean age 47 yr, 46% male) evaluated in our GI physiology laboratory. The indication for testing was recorded and characterized as esophageal or supraesophageal. Abnormal esophageal acid exposure was defined as a distal esophageal pH <4 for more than 4.2% of the total monitoring time (>6.3% upright, >1.2% supine) or a proximal esophageal acid exposure time of greater than 1.1% total (>1.3% upright, 0% supine). IEM was defined as more than two of 10 ineffective peristaltic waves. Seventy-two patients had esophageal-presenting symptoms, and 12 had supraesophageal symptoms. The prevalence of abnormal esophageal acid exposure was similar in patients with esophageal and supraesophageal symptoms (69% vs 92%, p = 0.17). Abnormal motility was identified in 26 patients (31%). IEM was the most common motility disturbance (77%, 20 patients). The frequency of motility disorders was similar in patients with and without abnormal esophageal acid exposure (30% vs 35%, p = 0.79), in patients with esophageal or supraesophageal symptoms (32% vs 25%, p = 0.75, for all patients; 30% vs 27%, p = 1.00, for patients with abnormal esophageal acid exposure), and among upright, supine, and combined refluxers (33%, 9%, and 35%, p = 0.26).

IEM does not stand alone as a significant marker for the presence of GERD in general or supraesophageal reflux disease in particular.

Does increased expression of sialyl-Lewis A correlate with poor survival in upper urinary tract urothelial cancer patients?

We have previously shown that both sialyl-Lewis A (sLe(a)) and sialyl-Lewis X (sLe(x)) antigens are involved in E-selectin-mediated adhesion of some urothelial cancer cells to the endothelium in vitro. The present study was undertaken to determine whether these antigens could serve as prognostic parameters. We immunohistochemically examined the expression of sLe(a) and sLe(x) in 90 human upper urinary tract urothelial tumours. Fifty-eight (64%) tumours were found to express sLe(a), while sixty-one (68%) expressed sLe(x). The expression between sLe(a) and sLe(x) was correlated positively (p < 0.0001). Neither sLe(a) nor sLe(x) expression was correlated with any other pathological parameter. The cause-specific survival rate was significantly worse in sLe(a)-positive patients than in sLe(a)-negative patients (5-year survival rates 64% and 93%, respectively: p = 0.023), while the survival rates were not statistically different between sLe(x)-positive and sLe(x)-negative patients. By Cox's multivariate analysis, sLe(a) expression as well as lymph node involvement and venous invasion was an independent risk value to predict survival (p < 0.05). In 24 high-risk patients having lymph node metastasis or having both venous invasion and sLe(a) expression, the 5-year survival rate was 29%, while it was 93% in the other 66 patients (p < 0.0001).

These results indicate that increased expression of sLe(a) antigen as well as lymph node metastasis and venous invasion may be associated with shorter cause-specific survival in upper urinary tract urothelial cancer. The combination of these factors offers a better prediction of prognosis.

Do short-term alcohol and drug treatment outcomes predict long-term outcome?

Although addiction is recognized as a chronic, relapsing condition, few treatment studies, and none in a commercially insured managed care population, have measured long-term outcomes. We examined the relationship of 6-month treatment outcomes to abstinence 5 years post-treatment, and whether the predictors of abstinence at 5 years were different for those who were, and were not, abstinent at 6 months. The sample (N=784) is from an outpatient (day hospital and traditional outpatient) managed care chemical dependency program. Subjects were interviewed at baseline, 6 months, and 5 years. Logistic regression analysis was used to assess which individual, treatment and extra-treatment characteristics predicted alcohol and drug abstinence at 5 years. Abstinence at 6 months was an important predictor of abstinence at 5 years. Among those abstinent at 6 months, predictors of abstinence at 5 years were older age, being female, 12-step meeting attendance, and recovery-oriented social networks. Among those not abstinent at 6 months, being alcohol dependent rather than drug dependent, 12-step meeting attendance, treatment readmission, and recovery-oriented social networks predicted abstinence at 5 years.

Our findings demonstrate a clear association between short-term and long-term treatment success. In addition, these results strongly support the importance of recovery-oriented social networks for those with good short-term outcomes, and the beneficial impact of readmission for those not initially successful in treatment.

Constitutive activation and accelerated maturation of peripheral blood T cells in healthy adults in Burkina Faso compared to Germany: the case of malaria?

It is not exactly known how frequent exposure to Plasmodium falciparum shapes the peripheral blood T-cell population in healthy West Africans. The frequency of peripheral blood CD4(+) lymphocytes responding to Plasmodium falciparum merozoite surface protein 1 (PfMSP-1) by production of interferon-gamma (IFN-γ), interleukin-2 (IL-2) or tumor necrosis factor-alpha (TNF-α) was determined using a commercially available flow cytometric activation assay (FastImmune) in 17 healthy adults in Nouna, Burkina Faso. T-cell activation and maturation in peripheral blood of healthy adults in Burkina Faso (n=40) and Germany (n=20) were compared using immunophenotyping and three-colour flow cytometry. Significant numbers of PfMSP-1 -specific CD4(+) lymphocytes producing IFN-γ, IL-2 and/or TNF-α were detected in 14 healthy adults in Nouna. Cytokine profiles showed predominant production of IFN-γ and TNF-α. Compared to Germans, Burkinabé showed markedly lower proportions of CCR7+ CD45RA+ naive CD4(+) cells and slightly higher frequencies of CD95(+)CD4(+) T-cells and of CD38(+) CD8(+) T-cells. The median antibody-binding capacity of CD95(dim) CD4(+) T-cells in Burkinabé was more than twice the value observed in Germans (263 vs. 108 binding sites per cell, p<0.0001).

We hypothesize that an IFN-γ-induced increase in the expression level of CD95 on CD4(+) lymphocytes may lower the activation threshold of resting naive CD4(+) T-cells in healthy adults living in Burkina Faso. Bystander activation of these cells deserves further study as a molecular mechanism linking strong IFN-γ responses against Plasmodium falciparum to decreased susceptibility to parasitemia observed in specific ethnic groups in West Africa.

Do extracellular acidification and hyperkalemia induce changes in HERG inhibition by ibutilide?

A high incidence of proarrhythmia has been reported with ibutilide, especially in patients with underlying heart diseases. Our previous studies have shown that extracellular acidosis and hyperkalemia attenuate the HERG-inhibitory effect of proarrhythmic drugs, e.g. quinidine, but have little impact on the less-proarrhythmic drug amiodarone. We hypothesized that ibutilide would behave like quinidine in the presence of extracellular acidosis and hyperkalemia. HERG was expressed on Xenopus oocytes, and the two-electrode voltage clamp technique was employed. Our results showed that ibutilide was a potent HERG inhibitor. When extracellular solution contained 5 mM KCl and pH was 7.4, the IC(50) of ibutilide was 0.9 +/- 0.1 microM. The inhibitory effect of ibutilide was attenuated when extracellular pH decreased to 6.2. There was a significant difference in current inhibition by ibutilide at pH 7.4 versus pH 6.2 (p < 0.01). When the extracellular potassium concentration was increased from 5 to 10 mM, ibutilide produced less current inhibition, and the IC(50) was increased to 2.0 +/- 0.1 microM.

Extracellular acidosis and hyperkalemia attenuate the HERG-inhibitory effect of ibutilide. The differences in HERG inhibition between acidic and hyperkalemic regions compared to normal regions in the myocardium may result in heterogeneity in repolarization, which may contribute to the proarrhythmic toxicity of ibutilide.

Does regional media coverage influence the public 's negative attitudes to policy implementation success in Sweden?

One central aspect of health literacy is knowledge of patients' rights. Being an important source of information about health and health care, the media may influence health literacy and act as a policy implementer. To investigate whether regional news media coverage in Sweden is linked to (i) the public's awareness and knowledge of a patient's rights policy, the waiting-time guarantee and (ii) the public's attitudes to how the guarantee's time limits are met, that is, implementation success. Three types of data are used. First, a national telephone survey of the public's awareness, knowledge and attitudes; second, media coverage information from digital media monitoring; and third, official waiting-time statistics. Bivariate and multivariate regression analyses are performed with the 21 Swedish county councils/regions as a base. In the county councils/regions, non-awareness ranged from 1 to 15% and knowledge from 47 to 67%. There are relatively large differences between population groups. The amount of regional media coverage shows no significant correlation to the level of awareness and knowledge. There is, however, a significant correlation to both positive and negative attitudes; the latter remains after controlling for actual waiting times.

At the national level, the media function as a policy implementer, being the primary source of information. At the regional level, the media are part of the political communication, reporting more extensively in county councils/regions where the population holds negative views towards the achievement in implementing the guarantee. We conclude that Swedish authorities should develop its communication strategies to bridge health literacy inequalities.

Is the outcome of caudal epidural injections affected by patient positioning?

A prospective, randomized controlled trial. To investigate the effect of the lateral decubitus position, after a caudal epidural injection, on outcome. Caudal epidural injections are used widely in the treatment of low back pain and radicular leg pain. Various measures have been used to improve the efficacy of these injections in previous studies. Our aim was to investigate the effect of the lateral decubitus position, after administering a caudal epidural injection, on outcome. Fifty-seven patients undergoing caudal epidural injection for low back pain associated with radicular leg pain were randomly allocated into 2 groups. Group 1 (treatment group) had 28 patients who were placed in the lateral decubitus position after injection. Group 2 (control group) had 29 patients who were laid supine after injection. Patients were assessed before injection using the Verbal Pain Score (VPS) and the Oswestry Disability Index (ODI). They were reassessed after 6 weeks using the same outcome measures. Both groups demonstrated improvement after injection. The degree of improvement in the VPS was significantly greater in group 1 compared with group 2 (P = 0.00007). The degree of improvement in the ODI was not statistically significant (P = 0.14).

Laying a patient on the side of their leg pain after a caudal epidural injection has a beneficial effect on the degree of pain relief. We recommend that this simple and safe maneuver be introduced routinely after administering a caudal epidural injection, to aid in the eventual outcome of a potentially difficult clinical problem.

Does waste water effluent contribute to the dissemination of CTX-M-15 in the natural environment?

Multidrug-resistant Enterobacteriaceae pose a significant threat to public health. We aimed to study the impact of sewage treatment effluent on antibiotic resistance reservoirs in a river. River sediment samples were taken from downstream and upstream of a waste water treatment plant (WWTP) in 2009 and 2011. Third-generation cephalosporin (3GC)-resistant Enterobacteriaceae were enumerated. PCR-based techniques were used to elucidate mechanisms of resistance, with a new two-step PCR-based assay developed to investigate bla(CTX-M-15) mobilization. Conjugation experiments and incompatibility replicon typing were used to investigate plasmid ecology. We report the first examples of bla(CTX-M-15) in UK river sediment; the prevalence of bla(CTX-M-15) was dramatically increased downstream of the WWTP. Ten novel genetic contexts for this gene were identified, carried in pathogens such as Escherichia coli ST131 as well as indigenous aquatic bacteria such as Aeromonas media. The bla(CTX-M-15) -gene was readily transferable to other Gram-negative bacteria. We also report the first finding of an imipenem-resistant E. coli in a UK river.

The high diversity and host range of novel genetic contexts proves that evolution of novel combinations of resistance genes is occurring at high frequency and has to date been significantly underestimated. We have identified a worrying reservoir of highly resistant enteric bacteria in the environment that poses a threat to human and animal health.

Do infants with colic have a normal sleep structure at 2 and 7 months of age?

To compare nighttime sleep structure between infants with colic and a control group. Sleep and cry times of 15 infants with colic and 16 infants in a control group were recorded with the use of a daily diary at the ages of 5 weeks and 6 months. The diary was kept at home for a 1-week period. Overnight polygraphic sleep recordings in a sleep laboratory were performed when the infants were 2 months of age and were repeated for 11 infants with colic and 14 infants in a control group at 7 months of age. Daily sleep time was shorter in infants with colic compared with the control group at 5 weeks of age (P =.001). Polygraphic data showed a similar sleep structure between the study groups at 2 and 7 months of age. Infants with colic had somewhat more obstructive apneas during rapid eye movement sleep at the age of 2 months (P =.04), and they had fewer awakenings at the age of 7 months than the control group (P =.003).

Infants with colic had normal sleep polygraphic finding at 2 and 7 months of age including sleep structure, movements, and breathing. Despite the shorter reported daily sleep times, the polygraphic data did not suggest infantile colic to be associated with a sleep disorder.

Does extracellular ATP inhibit twitching motility-mediated biofilm expansion by Pseudomonas aeruginosa?

Pseudomonas aeruginosa is an opportunistic pathogen that exploits damaged epithelia to cause infection. Type IV pili (tfp) are polarly located filamentous structures which are the major adhesins for attachment of P. aeruginosa to epithelial cells. The extension and retraction of tfp powers a mode of surface translocation termed twitching motility that is involved in biofilm development and also mediates the active expansion of biofilms across surfaces. Extracellular adenosine triphosphate (eATP) is a key "danger" signalling molecule that is released by damaged epithelial cells to alert the immune system to the potential presence of pathogens. As P. aeruginosa has a propensity for infecting damaged epithelial tissues we have explored the influence of eATP on tfp biogenesis and twitching motility-mediated biofilm expansion by P. aeruginosa. In this study we have found that eATP inhibits P. aeruginosa twitching motility-mediated expansion of interstitial biofilms at levels that are not inhibitory to growth. We have determined that eATP does not inhibit expression of the tfp major subunit, PilA, but reduces the levels of surface assembled tfp. We have also determined that the active twitching zone of expanding P. aeruginosa interstitial biofilms contain large quantities of eATP which may serve as a signalling molecule to co-ordinate cell movements in the expanding biofilm. The inhibition of twitching motility-mediated interstitial biofilm expansion requires eATP hydrolysis and does not appear to be mediated by the Chp chemosensory system.

Endogenous eATP produced by P. aeruginosa serves as a signalling molecule to co-ordinate complex multicellular behaviours of this pathogen. Given the propensity for P. aeruginosa to infect damaged epithelial tissues, our observations suggest that eATP released by damaged cells may provide a cue to reduce twitching motility of P. aeruginosa in order to establish infection at the site of damage. Furthermore, eATP produced by P. aeruginosa biofilms and by damaged epithelial cells may play a role in P. aeruginosa pathogenesis by inducing inflammatory damage and fibrosis. Our findings have significant implications in the development and pathogenesis of P. aeruginosa biofilm infections.

Are measures of perception of bronchoconstriction and clinical and functional data interrelated in asthma?

Sensitivity and absolute perceptual magnitude characterize the perception of bronchoconstriction (PB). To define whether clinical and functional characteristics and level of bronchial hyperresponsiveness (BHR) correlate with these two PB indexes during bronchial challenge in asthma. PB on both the Borg scale and the visual-analogue scale (VAS) was assessed in 45 consecutive asthmatics during a methacholine-induced decrease in forced expiratory volume in 1 s (FEV(1)) and specifically quantified as Borg and VAS slope, as a measure of sensitivity, whereas scores at a 20% FEV(1) decrease (PB(20)) were assessed as a measure of absolute perceptual magnitude. Clinical score and BHR were also assessed. PB(20) related to slope on both the Borg scale and the VAS (p < 0.0001). PB(20) and slope related neither to clinical score nor to baseline functional data on both scales. The relationship between the level of BHR and PB(20) on either scale was of questionable clinical significance (r(2) = 7%).

Irrespective of the scale employed, our data indicate the need for directly assessing PB rather than deriving it from clinical and functional data and level of BHR.

Does cholecystokinin stimulation lead to increased oxytocin secretion in women?

To find out if cholecystokinin (CCK) stimulates the secretion of oxytocin in humans, and if there are any differences in secretion between healthy women and those with normal-transit constipation. Prospective open study. Teaching hospital, Sweden. 8 healthy female volunteers and 6 women with chronic refractory normal-transit constipation. Subjects were fasted before experiments. On one day they were given emulsified corn oil and another an intravenous injection of 1 Ivy dog unit (IDU) CCK/kg body weight. Blood samples were taken before each experiment at 10 minutes and at the time the experiments started. Blood samples were also taken after each experiment at 10, 20, 30, 45, 60, 90 and 120 minutes. Concentrations of CCK and oxytocin. Ingestion of corn oil significantly increased the plasma concentration of CCK in both groups (healthy women p = 0.03 and constipated women p = 0.008). Injection of CCK also led as expected to hypercholecystokininaemia in both groups (p = 0.008 and p = 0.03, respectively). The corn oil increased oxytocin secretion in both groups (p = 0.02 and 0.03, respectively) and exogenous CCK increased the secretion still further (p = 0.008 and 0.03, respectively).

Both corn oil and injection of CCK led to an increased CCK concentration in plasma. Oxytocin was secreted in response to endogenous as well as exogenous CCK stimulation. There was no difference between healthy and constipated women in either parameter analysed.

Are clinical and serum-based markers associated with death within 1 year of de novo implant in primary prevention ICD recipients?

Implantable cardioverter-defibrillator (ICD) implantation is contraindicated in those with <1-year life expectancy. The aim of this study was to develop a risk prediction score for 1-year mortality in patients with primary prevention ICDs and to determine the incremental improvement in discrimination when serum-based biomarkers are added to traditional clinical variables. We analyzed data from the Prospective Observational Study of Implantable Cardioverter-Defibrillators, a large prospective observational study of patients undergoing primary prevention ICD implantation who were extensively phenotyped for clinical and serum-based biomarkers. We identified variables predicting 1-year mortality and synthesized them into a comprehensive risk scoring construct using backward selection. Of 1189 patients deemed by their treating physicians as having a reasonable 1-year life expectancy, 62 (5.2%) patients died within 1 year of ICD implantation. The risk score, composed of 6 clinical factors (age ≥75 years, New York Heart Association class III/IV, atrial fibrillation, estimated glomerular filtration rate <30 mL/min/1.73 m(2), diabetes, and use of diuretics), had good discrimination (area under the curve 0.77) for 1-year mortality. Addition of 3 biomarkers (tumor necrosis factor α receptor II, pro-brain natriuretic peptide, and cardiac troponin T) further improved model discrimination to 0.82. Patients with 0-1, 2-3, 4-6, or 7-9 risk factors had 1-year mortality rates of 0.8%, 2.7%, 16.1%, and 46.2%, respectively.

Individuals with more comorbidities and elevation of specific serum biomarkers were at increased risk of all-cause mortality despite being deemed as having a reasonable 1-year life expectancy. A simple risk score composed of readily available clinical data and serum biomarkers may better identify patients at high risk of early mortality and improve patient selection and counseling for primary prevention ICD therapy.

Is There a Correlation between the Number of Follicular Flushings, Oocyte/Embryo Quality and Pregnancy Rate in Assisted Reproductive Technology Cycles?

To determine the mean number of follicular flushings needed to retrieve the maximal number of oocytes and to investigate the correlation between the number of flushings and oocyte/embryo quality as well as reproductive outcome. This prospective study included 154 oocyte retrieval cycles in 138 patients undergoing assisted reproductive technology treatment. During oocyte retrieval, aspirate and flushes were collected in four separate collections (A, F1, F2, F3) and inspected for the presence of an oocyte-cumulus complex (OCC). Outcome variables included oocyte recovery rate, percentage of mature oocytes, fertilization rate, day 2/day 3/day 5 embryo quality and clinical pregnancy rate/live birth rate. Out of 1,495 OCCs collected, 91% were obtained in collections A or F1. Significantly more mature oocytes were obtained in collection A (p = 0.03). The fertilization rate was similar for oocytes obtained in the four separate collections (p = 0.50). The proportion of good-quality day 2/day 3/day 5 embryos was similar (p = 0.93, p = 0.85 and p = 0.14, respectively). Clinical pregnancy rate and live birth rate were not significantly affected by the first two flushes. No live birth emanated from oocytes obtained from the third flush onwards.

More than 90% of all recovered OCCs were obtained after follicular aspiration followed by follicular flushing up to two times (collections A and F1). Follicular flushing may increase the oocyte recovery rate and does not have a negative influence on fertilization rate, day 2/day 3/day 5 embryo quality or pregnancy rate.

Does acute tryptophan depletion attenuate conscious appraisal of social emotional signals in healthy female volunteers?

Acute tryptophan depletion (ATD) decreases levels of central serotonin. ATD thus enables the cognitive effects of serotonin to be studied, with implications for the understanding of psychiatric conditions, including depression. To determine the role of serotonin in conscious (explicit) and unconscious/incidental processing of emotional information. A randomized, double-blind, cross-over design was used with 15 healthy female participants. Subjective mood was recorded at baseline and after 4 h, when participants performed an explicit emotional face processing task, and a task eliciting unconscious processing of emotionally aversive and neutral images presented subliminally using backward masking. ATD was associated with a robust reduction in plasma tryptophan at 4 h but had no effect on mood or autonomic physiology. ATD was associated with significantly lower attractiveness ratings for happy faces and attenuation of intensity/arousal ratings of angry faces. ATD also reduced overall reaction times on the unconscious perception task, but there was no interaction with emotional content of masked stimuli. ATD did not affect breakthrough perception (accuracy in identification) of masked images.

ATD attenuates the attractiveness of positive faces and the negative intensity of threatening faces, suggesting that serotonin contributes specifically to the appraisal of the social salience of both positive and negative salient social emotional cues. We found no evidence that serotonin affects unconscious processing of negative emotional stimuli. These novel findings implicate serotonin in conscious aspects of active social and behavioural engagement and extend knowledge regarding the effects of ATD on emotional perception.

Is [ Lateral trunk control in a sitting test associated with mobility and Instrumental Activities of Daily Living among community-dwelling elderly people ]?

We hypothesized that during walking, mediolateral instability with gait disorders could be reflected in the characteristics of lateral trunk control while sitting. We investigated the association between lateral trunk control while sitting, mobility and Instrumental Activities of Daily Living (IADL) among community-dwelling elderly people. A cross-sectional analysis was carried out of the data of 33 men and 102 women in a community-dwelling elderly population (average age, 73.2±6.0 years). A Seated Side-Tapping test was developed to assess lateral trunk control while sitting. We used the Lawton IADL scale, the 5-m normal walking test and the Timed "Up & Go" test (TUG) to measure mobility. The mean duration of the Seated Side-Tapping test was 5.0±0.9 seconds, ranging from 3.1 to 8.2 seconds. The test was normally distributed (p=0.200). The Seated Side-Tapping test was significantly associated with TUG, after controlling for age (r=0.46). The group with disability in at least 1 IADL item was significantly slower in the Seated Side-Tapping test, especially in men (5.7 sec for men, 5.5 sec for women vs. the independent group, 4.8 sec for both). Only the Seated Side-Tapping test remained significantly associated with IADL disability after logistic regression analysis.

These results indicate that lateral trunk control in sitting is associated with mobility and disability in at least 1 item of the IADL. These results support the potential use of the Seated Side-Tapping test to safely carry out risk assessments for frail elderly patients.

Does spatial variation of insecticide resistance in the dengue vector Aedes aegypti present unique vector control challenges?

Dengue is a major public health problem in Mexico, where the use of chemical insecticides to control the principal dengue vector, Aedes aegypti, is widespread. Resistance to insecticides has been reported in multiple sites, and the frequency of kdr mutations associated with pyrethroid resistance has increased rapidly in recent years. In the present study, we characterized patterns of insecticide resistance in Ae. aegypti populations in five small towns surrounding the city of Merida, Mexico. A cross-sectional, entomological survey was performed between June and August 2013 in 250 houses in each of the five towns. Indoor resting adult mosquitoes were collected in all houses and four ovitraps were placed in each study block. CDC bottle bioassays were conducted using F0-F2 individuals reared from the ovitraps and kdr allele (Ile1016 and Cys1534) frequencies were determined. High, but varying, levels of resistance to chorpyrifos-ethyl was detected in all study towns, complete susceptibility to bendiocarb in all except one town, and variations in resistance to deltamethrin between towns, ranging from 63-88% mortality. Significant associations were detected between deltamethrin resistance and the presence of both kdr alleles. Phenotypic resistance was highly predictive of the presence of both alleles, however, not all mosquitoes containing a mutant allele were phenotypically resistant. An analysis of genotypic differentiation (exact G test) between the five towns based on the adult female Ae. aegypti collected from inside houses showed highly significant differences (p < 0.0001) between genotypes for both loci. When this was further analyzed to look for fine scale differences at the block level within towns, genotypic differentiation was significant for both loci in San Lorenzo (Ile1016, p = 0.018 and Cys1534, p = 0.007) and for Ile1016 in Acanceh (p = 0.013) and Conkal (p = 0.031).

The results from this study suggest that 3 years after switching chemical groups, deltamethrin resistance and a high frequency of kdr alleles persisted in Ae. aegypti populations. The spatial variation that was detected in both resistance phenotypes and genotypes has practical implications, both for vector control operations as well as insecticide resistance management strategies.

Is negative chronotropic response to low-dose atropine associated with parasympathetic nerve-mediated cardiovascular response in postoperative patients with congenital heart disease?

To investigate the negative chronotropic response (NCR) to low-dose atropine in postoperative patients with congenital heart disease (CHD). Low-dose atropine causes a NCR through the central nervous system muscarinic receptor and is attenuated in adult heart failure patients. It has never been evaluated in CHD patients. NCR corrected for basal heart rate (HR) (minimal HR/basal HR=cNCR) was determined after low-dose atropine (3 microg/kg) administration in 124 postoperative CHD patients (97 biventricular repair and 27 Fontan patients) and 11 controls and was compared with the cardiac autonomic nervous and functional status. The cNCR in simple CHD (post atrial or ventricular septal defect closure), complex biventricular CHD, and Fontan patients were 0.92+/-0.04, 0.94+/-0.04 and 0.96+/-0.04, respectively, and higher than in controls (0.87+/-0.03, p<0.001). In the complex CHD patients, higher cNCR was mainly associated with the lower pharmacologically determined cardiac parasympathetic nervous tone (PST), HR variability, high atrial natriuretic peptide, and lower right ventricular ejection fraction (p<0.0001). In Fontan patients, the lower beta sensitivity of the sinus node and the PST mainly determined the higher cNCR (p<0.01) and the cNCR did not correlate with either hemodynamics or exercise capacity.

NCR is attenuated in proportion to the impaired cardiac parasympathetic nervous system and hemodynamics in postoperative complex biventricular CHD patients. In addition to PST, beta sensitivity of the sinus node significantly influences the NCR in Fontan patients.

Does chronic intermittent hypobaric hypoxia ameliorate diabetic nephropathy through enhancing HIF1 signaling in rats?

Our previous study demonstrated that chronic intermittent hypobaric hypoxia (CIHH) had anti-diabetes effect. The present study was to explore the renal protective effect of CIHH in diabetic rats. Sprague-Dawley rats were randomly divided into three groups: diabetes mellitus group (DM, induced by high-fat diet combined with low-dose streptozotocin), diabetes plus CIHH treatment group (DM+CIHH, simulated 5000-m altitude, 6h per day for 28days, after diabetes model confirmed) and control group (CON). Systolic arterial blood pressure (SAP), blood biochemicals, urinary albumin, and histopathology of kidney were determined. The superoxide dismutase (SOD) activity, malondialdehyde (MDA) level, protein levels of hypoxia induced factors (HIFs) and transforming growth factor β1 (TGF-β1) in kidney were assayed. The increased SAP, urinary albumin, hyperplasia of glomerular, fibrosis in mesangial and glomerular, and abnormal lipid metabolism in diabetic rats were ameliorated by CIHH treatment. And decreased superoxide dismutase (SOD) activity and increased malondialdehyde (MDA) level in diabetic kidney were reversed in CIHH-treated DM rats. In addition up-regulated TGF-β1 and down-regulated HIF1α in diabetic kidney returned back to normal level in CIHH-treated DM rats.

These data demonstrated for the first time that CIHH had protective effects against the early stage damage of diabetic nephropathy through activating HIF1 signaling, improving anti-oxidation and inhibiting TGF-β1 signaling in diabetic rats.

Is cryoablative response of prostate cancer cells influenced by androgen receptor expression?

To investigate in prostate cancer cells the consequences of androgen-insensitivity (AI) development on the cellular and molecular responses to freezing, as a challenge in prostate cancer treatment occurs when the androgen-sensitive (AS) phenotype switches to an AI phenotype, the latter of which is often refractory to many therapies. PC-3 (AI) and LNCaP (AS) were each genetically altered to express the opposite phenotype and subjected to an in vitro freezing model. Viability, caspase inhibitor and Western blot studies were used to determine the basis of the differential responses of AI and AS cells. LNCaP high-passage cells, formed by repeated passage of LNCaP (AS) cells, were AI and showed a phenotypic shift to freeze resistance matching the freeze response of PC-3 cells (AI). While stably transfected androgen receptor (AR)-transfected cells (PC-3 AR) had a freezing sensitivity similar to that of the LNCaP (AS) cell line. Importantly, AI cell lines survived and recovered from freezing exposure to temperatures as low as -40 degrees C whereas AS cell lines did not. Caspase inhibition studies and related fluorescent probes showed an elevated level of apoptotic involvement in both AS cell lines after freezing compared with their AI counterparts. Western blot analysis showed that AR expression was modified after exposure to freezing.

This study suggests that AS cancers may be far more sensitive to a freezing insult and this might be linked to elevated apoptosis and caspase activity. As such, cryoablation may prove most effective in cancer cells that have not yet progressed to a more resistant AI phenotype, but both generic variants can be fully ablated at sufficiently low temperatures.

Is packed red blood cell transfusion an independent risk factor for necrotizing enterocolitis in premature infants?

To determine whether a temporal association exists between antecedent packed red blood cell (PRBC) transfusions and necrotizing enterocolitis (NEC) in premature infants. This case-control study included inborn infants from a single center who developed NEC during a 2-year period. For every NEC infant, two matched controls from the same period were chosen based on gestational age and birth weight. Transfusion-related NEC was defined as antecedent PRBC transfusion within 48 h prior to the onset of any symptoms attributable to NEC. Bivariate analyses were used to compare baseline characteristics of all infants. To determine the raw odds ratio for the presence of exposure (transfusion) versus outcome (NEC), the hospital course (ages 6 to 63 days) of all study infants was divided into 48-h epochs; occurrence of transfusion and NEC was noted within each epoch. Generalized estimating equations were used to estimate the adjusted odds for developing NEC within an epoch with and without antecedent transfusion, controlling chronological age within infant as well as for gestational age, gender, feeding status in prior 48-h epoch and indicators of disease severity. There were 3652 48-h epochs and 557 transfusions among 49 NEC infants and 97 controls; 17 infants had transfusion-related NEC, yielding a raw odds ratio of 3.01 (P<0.001). The adjusted odds ratios were 2.97 (P=0.003) for transfusion and 2.76 (P=0.05) for feeding status in the prior 48-h epoch. Infants who were being fed in the 48-h period prior to transfusion were more than eight times more likely to develop NEC than infants who were neither fed nor transfused.

Antecedent PRBC transfusion appears to be an independent risk factor for developing NEC during the subsequent 48-h period.

Are micturating cystourethrograms necessary for all cases of antenatally diagnosed hydronephrosis?

Since 1995 we have, at our centre, adopted a selective approach to performing micturating cystourethrograms (MCUGs) on patients with antenatally diagnosed hydronephrosis. This study reviews the outcome of this policy. We carry out MCUGs only if any of the following features are present on ultrasound: bilateral hydronephrosis, ureteric dilatation, renal scarring, bladder wall thickness greater than 5mm, or presence of a duplex system or ureterocele. Patients with simple unilateral hydronephrosis are excluded, and are managed with 6 months' trimethoprim prophylaxis and ultrasound surveillance with a minimum of 3 years' follow up. Fifty-five patients were referred with an antenatal diagnosis of hydronephrosis between 1999 and 2002; 26 (47%) did not have an MCUG. Of these, five had increasing hydronephrosis and required surgery for pelvi-ureteric junction obstruction, and three had a multicystic dysplastic kidney on postnatal scanning. In the remaining 18 patients, the hydronephrosis resolved spontaneously, with no renal scars or asymmetry. During follow up, none of these patients had a urinary tract infection.

We believe that vesico-ureteric reflux in most antenatally diagnosed hydronephrotic kidneys is physiological rather than pathological, and resolves with time without causing long-term renal damage. This is a separate entity from, rather than a precursor of, the pathological symptomatic refluxing kidney in older, mainly female children. Taking a more conservative approach to the postnatal investigation of antenatally diagnosed hydronephrotic kidneys has not resulted in any missed damaged kidneys, but has reduced the number of invasive investigations performed. A careful protocol and detailed postnatal ultrasonography are important to prevent missed pathological cases.

Is cardiac troponin T elevation at dialysis initiation associated with all-cause and cardiovascular mortality on dialysis in patients without diabetic nephropathy?

It is not known whether asymptomatic cardiac troponin T (cTnT) elevation is associated with all-cause or cardiovascular mortality in non-diabetic and advanced chronic kidney disease (CKD) patients. We measured cTnT in 248 consecutive patients at 1-2 weeks before dialysis initiation between March 2005 and August 2010 and followed them prospectively. A Cox proportional hazard model was used to investigate the relationship between cTnT and all-cause and cardiovascular mortality on dialysis. The median age of the patients was 67 years (male 59.3 %), and the prevalence of diabetic nephropathy (DN) was 38.3 %. Asymptomatic cTnT elevation (>0.01 ng/mL) was observed in 196 (79 %) and 111 (73 %) patients among the overall patients and among patients without DN, respectively. A total of 51 patients died during a median follow-up period of 31.6 months. The cTnT level was associated with all-cause [hazard ratio (HR) 1.453; 95 % confidence interval (CI) 1.093-1.931; P = 0.010] and cardiovascular mortality [HR 1.973; 95 % CI 1.127-3.454; P = 0.017] on dialysis after extensive adjustment in the overall patient population. Patients without DN showed similar associations as those for the overall patient population (all-cause mortality: HR 1.566; 95 % CI 1.048-2.339; P = 0.029 and cardiovascular mortality: HR 2.657; 95 % CI 1.115-6.328; P = 0.027).

Asymptomatic cTnT elevation might be strongly associated with all-cause and cardiovascular mortality in patients without DN, as well as in the overall advanced CKD patients. We suggest that cardiovascular risk in patients with pre-dialysis CKD should be stratified according to cTnT levels.

Does fetal striatal grafting slow motor and cognitive decline of Huntington 's disease?

To assess the clinical effect of caudate-putaminal transplantation of fetal striatal tissue in Huntington's disease (HD). We carried out a follow-up study on 10 HD transplanted patients and 16 HD not-transplanted patients. All patients were evaluated with the Unified HD Rating Scale (UHDRS) whose change in motor, cognitive, behavioural and functional capacity total scores were considered as outcome measures. Grafted patients also received morphological and molecular neuroimaging. Patients were followed-up from disease onset for a total of 309.3 person-years (minimum 5.3, median 11.2 years, maximum 21.6 years). UHDRS scores have been available since 2004 (median time of 5.7 years since onset, minimum zero, maximum 17.2 years). Median post-transplantation follow-up was 4.3 years, minimum 2.8, maximum 5.1 years. Adjusted post-transplantation motor score deterioration rate was reduced compared to the pretransplantation period, and to that of not-transplanted patients by 0.9 unit/years (95% CI 0.2 to 1.6). Cognitive score deterioration was reduced of 2.7 unit/years (95% CI 0.1 to 5.3). For grafted patients the 2-year post-transplantation [(18)F]fluorodeoxyglucose positron emission tomography (PET) showed striatal/cortical metabolic increase compared to the presurgical evaluation; 4-year post-transplantation PET values were slightly decreased, but remained higher than preoperatively. [(123)I]iodobenzamide single photon emission CT demonstrated an increase in striatal D2-receptor density during postgrafting follow-up.

Grafted patients experienced a milder clinical course with less pronounced motor/cognitive decline and associated brain metabolism improvement. Life-time follow-up may ultimately clarify whether transplantation permanently modifies the natural course of the disease, allowing longer sojourn time at less severe clinical stage, and improvement of overall survival.

Informed consent in pediatric surgery: Do parents understand the risks?

To investigate parent understanding of the risks of pediatric ear, nose, and throat surgery after counseling with and without the use of informational aids. Prospective, randomized trial. Academic tertiary care center. Parents of children undergoing ear, nose, and throat surgery. Parents were randomized to receive standard informed consent with or without detailed informational aids. Parents completed identical questionnaires testing their general procedure knowledge and their recall of 9 specific surgical risks both immediately after counseling and on the day of surgery. Thirty-four parents enrolled in and completed the study (18 in the control group and 16 in the test group). The mean time from informed consent to surgery was 6.3 days (range, 1-22 days). Parents in the test group scored significantly higher on identifying the 9 risks on both the preoperative questionnaire (mean score, 6.00 vs 4.44; P = .007, 2-tailed t test) and the postoperative questionnaire (6.25 vs 4.17; P<.001). There was a negative correlation (inverse relationship) between parent education score and risk recall, with parents with lower education levels scoring higher on both the preoperative (Pearson r = -0.36; P = .04) and the postoperative (r = -0.35; P = .04) surveys. The maternal parent recalled risks significantly better than the paternal parent, with surgical risk recall scores of 5.46 out of 9 vs 3.67 out of 9 (P = .02, 2-tailed t test).

Parents of children undergoing ear, nose, and throat surgery recall far less than 100% of counseled risks. The use of detailed surgical risk counseling improves measured parental understanding of surgical risk. Parental educational level and maternal vs paternal parent may affect risk counseling recall.

Is non-cladding optical fiber available for detecting blood or liquids?

Serious accidents during hemodialysis such as an undetected large amount of blood loss are often caused by venous needle dislodgement. A special plastic optical fiber with a low refractive index was developed for monitoring leakage in oil pipelines and in other industrial fields. To apply optical fiber as a bleeding sensor, we studied optical effects of soaking the fiber with liquids and blood in light-loss experimental settings. The non-cladding optical fiber that was used was the fluoropolymer, PFA fiber, JUNFLON™, 1 mm in diameter and 2 m in length. Light intensity was studied with an ordinary basic circuit with a light emitting source (880 nm) and photodiode set at both terminals of the fiber under certain conditions: bending the fiber, soaking with various mediums, or fixing the fiber with surgical tape. The soaking mediums were reverse osmosis (RO) water, physiological saline, glucose, porcine plasma, and porcine blood. The light intensities regressed to a decaying exponential function with the soaked length. The light intensity was not decreased at bending from 20 to 1 cm in diameter. The more the soaked length increased in all mediums, the more the light intensity decreased exponentially. The means of five estimated exponential decay constants were 0.050±0.006 standard deviation in RO water, 0.485±0.016 in physiological saline, 0.404±0.022 in 5% glucose, 0.503±0.038 in blood (Hct 40%), and 0.573±0.067 in plasma. The light intensity decreased from 5 V to about 1.5 V above 5 cm in the soaked length in mediums except for RO water and fixing with surgical tape.

We confirmed that light intensity significantly and exponentially decreased with the increased length of the soaked fiber. This phenomena could ideally, clinically be applied to a bleed sensor.

Is microglial activation involved in morphine tolerance mediated by toll-like receptor 4?

Morphine is a powerful analgesic but its effect is often diminished owing to the development of tolerance. It has been suggested that morphine activates microglia through its action on the toll-like receptor 4 (TLR4) in the spinal cord, leading to suppression of the morphine effect. However, it has not been examined whether the development of morphine tolerance is affected by the deletion and mutation of the TLR4 gene. Mice were treated with morphine (60 mg/kg) or vehicle once daily for five consecutive days to induce morphine tolerance, which was assessed by the tail-flick test before and after the treatment period. The effect of the microglial inhibitor minocycline, and the effect of TLR4 mutation (C3H/HeJ mouse) and deletion (TLR4-knockout mouse) on the development of morphine tolerance were tested. The expression of the microglial activation marker, CD11b, in the spinal cords of TLR4-knockout and wild-type mice after morphine treatment for 5 days was assessed by reverse-transcription polymerase chain reaction. Minocycline attenuated the development of morphine tolerance in mice. Mutation or deletion of the TLR4 gene did not significantly affect the development of morphine tolerance. CD11b mRNA expression was increased after morphine treatment both in TLR4-knockout and wild-type mice.

Microglial activation caused by a mechanism independent of TLR4 is involved in the development of morphine tolerance. Further studies are necessary to clarify the cellular mechanisms of morphine-induced microglial activation.

Do gATA3 mutations define a unique subtype of luminal-like breast cancer with improved survival?

The GATA3 gene (GATA-binding protein 3) is one of the most frequently mutated genes in breast cancer. The objective of the current study was to determine the clinicopathologic characteristics of patients with breast cancer harboring GATA3 mutations. The authors examined the somatic mutation status of GATA3 and performed survival analysis in The Cancer Genome Atlas (TCGA) cohort (n=934) and the Fudan University Shanghai Cancer Center (FUSCC) cohort (n=308). Patient characteristics, including age; menopausal status; tumor laterality; tumor size; lymph node status; tumor grade; molecular subtypes; adjuvant radiotherapy, chemotherapy, and endocrine therapy; and prognosis, together with PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha) and TP53 (tumor protein p53) mutation status, were collected. GATA3 mutations were detected in 8.8% of patients (82 of 934 patients) in the TCGA cohort and 14.9% of patients (46 of 308 patients) in the FUSCC cohort. GATA3 mutations were found to be significantly associated with luminal-like breast cancer (P=.002 in the TCGA cohort and P<.001 in the FUSCC cohort), and were highly mutually exclusive to PIK3CA mutations (P=.001 in the TCGA cohort and P=.003 in the FUSCC cohort) and TP53 mutations (P<.001 in both cohorts). Furthermore, GATA3 mutations were correlated with improved overall survival in the entire population (P=.025 in the TCGA cohort and P = .043 in the FUSCC cohort) as well as in patients with luminal-like disease who received adjuvant endocrine therapy.

GATA3 mutations mainly occur in patients with luminal-like breast cancer and have identifiable clinicopathologic and genetic characteristics, highlighting a subgroup of patients with breast cancer in whom limited therapy may be appropriate.

Anal physiology testing in fecal incontinence: is it of any value?

The prognostic value of postoperative manometry in fecal incontinence is still controversial. The aims of this study were to establish if Fecal Incontinence Severity Index (FISI) and Fecal Incontinence Quality of Life Scale (FIQL) scores correlate with anal manometry and endoanal ultrasound findings and to define if there is any prognostic value in performing anal manometry after patients are surgically treated for fecal incontinence. Fifty-three patients, all women, were identified. All patients underwent a surgical procedure and were analyzed pre- and postoperatively. Fecal incontinence was assessed using the FISI and FIQL. Patients who did not have these score were excluded. Manometry and ultrasound findings before treatment and manometry findings after treatment were compared with surgical patient's incontinence scores. Anal canal length was noted, and its association with the pre- and postoperative manometry finding and incontinence scores were compared. No correlation of pre- and postoperative resting and squeeze pressures with incontinence scores was found. Ultrasound findings had no correlation with manometry results and incontinence scores. Anal canal length correlated with both pre- and postoperative manometry findings but not with incontinence scores.

Preoperative anal manometry and endoanal ultrasound help in guiding treatment options in patients with fecal incontinence. A decrease in FISI and increase in FIQL scores after a sphincter repair quantifies improvement after incontinence surgery, while changes in anal manometry pressures readings do not.

Do aCTN3 R577X genotypes associate with Class II and deepbite malocclusions?

α-Actinins are myofibril anchor proteins that influence the contractile properties of skeletal muscles. ACTN2 is expressed in slow type I and fast type II fibers, whereas ACTN3 is expressed only in fast fibers. ACTN3 homozygosity for the 577X stop codon (ie, changing 577RR to 577XX, the R577X polymorphism) results in the absence of α-actinin-3 in about 18% of Europeans, diminishes fast contractile ability, enhances endurance performance, and reduces bone mass or bone mineral density. We have examined ACTN3 expression and genetic variation in the masseter muscle of orthognathic surgery patients to determine the genotype associations with malocclusion. Clinical information, masseter muscle biopsies, and saliva samples were obtained from 60 subjects. Genotyping for ACTN3 single nucleotide polymorphisms, real-time polymerase chain reaction quantitation of muscle gene message, and muscle morphometric fiber type properties were compared to determine statistical differences between genotype and phenotype. Muscle mRNA expression level was significantly different for ACTN3 single nucleotide polymorphism genotypes (P <0.01). The frequency of ACTN3 genotypes was significantly different for the sagittal and vertical classifications of malocclusion, with the clearest association being elevated 577XX genotype in skeletal Class II malocclusion (P = 0.003). This genotype also resulted in significantly smaller diameters of fast type II fibers in masseter muscles (P = 0.002).

ACTN3 577XX is overrepresented in subjects with skeletal Class II malocclusion, suggesting a biologic influence during bone growth. ACTN3 577XX is underrepresented in subjects with deepbite malocclusion, suggesting that muscle differences contribute to variations in vertical facial dimensions.

Is technology assisted guided self-help successful in treating female adolescents with bulimia nervosa?

This study aims to evaluate the long-term outcome of new technology assisted guided self-help in adolescents with bulimia nervosa (BN). One hundred and twenty-six patients with BN (29 adolescents and 97 adults) were randomly allocated to a cognitive behavioural therapy-based self-help program delivered by the Internet or bibliotherapy, both accompanied by e-mail guidance. Outcomes were assessed at baseline, month 4, 7 and 18 including remission rates and eating disorder associated psychopathology. In all, 44% of adolescents vs. 38.7% of adults were in remission at month 7, and 55% of adolescents vs. 62.5% of adults were in remission at follow-up. Objective binge eating and compensatory behaviour improved significantly over time in both groups, with the highest decrease during the first 4 months. A significant decrease over time and no group differences have been found in almost all EDI-2 subscales.

E-mail guided self-help (delivered via the Internet or bibliotherapy) is equally effective for adolescents as for adults with BN, and can be recommended as an initial step of treatment for this younger age group.

Is a modified breathing exercise program for asthma easy to perform and effective?

Breathing exercises are used by some asthmatic patients, yet are often difficult to perform and time-consuming. This study evaluated a simple, modified breathing exercise program regarding ease to perform and effectiveness as an adjunctive therapy. Subjects age 18 to 65 with a current diagnosis of persistent asthma were enrolled. A program that incorporated three different breathing exercises (yoga pranayama techniques, diaphragmatic breathing and pursed lip breathing) was taught to subjects. The program was designed to be completed in less than 10 minutes per day. Subjects completed the Asthma Control Test (ACT) and mini-Asthma Quality of Life Questionnaire (AQLQ) at baseline and at 1-month follow-up. They also completed a survey that asked them to rate the effectiveness and difficulty of the exercises, and whether they would recommend them in the future. A total of 74 subjects were enrolled in this study. The intervention improved breathing for 52.9% of the subjects, while 67.6% felt that their daily activity was improved and 66.1% noted that the exercises allowed decreased use of a rescue inhaler. Most subjects (80.9%) recommended breathing exercises as a complementary therapy for asthma and 79.4% of the subjects stated the exercises took less than 10 minutes per day total. Overall, ACT scores improved significantly (p = 0.002) with a statistically non-significant improvement in AQLQ scores.

A simple program of breathing exercises was found to be effective and could be completed in less than 10 minutes per day. Furthermore, there was a statistically significant improvement in ACT scores post-exercise.

Is it worth screening for vancomycin-resistant Enterococcus faecium colonization?

The screening of critically ill patients at high risk of vancomycin resistant enterococci (VRE) colonization, to detect and isolate colonized patients, is recommended to prevent and control the transmission of VRE. Screening asymptomatic carriers brings financial burden for institutions. In this study, we performed risk analysis for VRE colonization and determined the financial burden of screening in a middle-income country, Turkey. We retrospectively analyzed the VRE surveillance data from a pediatric hospital between 2010 and 2014. A case-control study was conducted to identify the risk factors of colonization. Total cost of VRE screening and additional costs for a VRE colonized patient (including active surveillance cultures and contact isolation) were calculated. During the 4-year period, 6,372 patients were screened for perirectal VRE colonization. The rate of culture-positive specimens among all patients screened was 239 (3.75%). The rate of VRE infection was 0.04% (n = 3) among all patients screened. Length of hospital stay, malignancy, and being transferred from another institution were independently associated risk factors for colonization. Annual estimated costs for the laboratory were projected as $19,074 (76,295/4) for all patients screened. Cost of contact isolation for each patient colonized in a ward and an intensive care unit was $270 and $718, respectively.

In developing countries, institutions should identify their own high-risk patients; screening priorities should be based on prevalence of infection and hospital financial resources.

Is gallstone disease associated with rectal cancer : a meta-analysis?

Increased levels of secondary bile acids after cholecystectomy and cholelithiasis are believed to increase the risk of colorectal cancer, and several studies have suggested that the risk of colorectal cancer may be the greatest proximally. Numerous conflicting studies have been published and it remains unclear whether the risk is apparent in the rectum. This meta-analysis aims to determine the risk of developing rectal cancer following gallstone disease or cholecystectomy. The prospective protocol included a literature search of PubMed, MEDLINE, EMBASE, and Current Contents (1950-2011). Selection criteria were developed to sort for studies exploring the relationship between cholelithiasis, cholecystectomy, and rectal cancer in an adult population. A random-effects model was used to generate pooled odds ratios (OR) and 95% confidence intervals (CI). Publication bias and heterogeneity were assessed. Of the 2358 studies identified, 42 were suitable for final analysis. There were 1,547,506 subjects in total, 14,226 diagnosed with rectal cancer, and 496,552 with gallstones or cholecystectomy. There was a statistically significant risk of rectal cancer following cholelithiasis (OR = 1.33; 95% CI = 1.02-1.73), though no risk was apparent following cholecystectomy (OR = 1.14; 95% CI = 0.92-1.41).

Cholelithiasis increases the risk of rectal cancer. No association exists between cholecystectomy and rectal cancer.

Do second-order stimuli always increase overall response rates in second-order schedules of reinforcement in the rat?

Second-order schedules of reinforcement have been used extensively to model reward-seeking and drug-seeking behaviour. Second-order stimuli within second-order schedules have been shown to enhance response rates during operant responding for natural reinforcers and drug reinforcers. This has led some to view second-order schedules of drug reinforcement as a model maintained of drug-seeking in addicts by drug-associated stimuli. However, the functional role of the second-order stimulus within second-order schedules is complex. We investigated the role of second-order stimuli within a second-order schedule of reinforcement [FI 4 min (FR10: S)] maintained by sweetened water reinforcement. Eight rats were trained to press a bar on a second-order schedule of reinforcement and tested in the presence and absence of the second-order stimulus. In contrast to most previous work, overall bar-pressing rates were significantly increased when the second-order stimulus was omitted (second-order stimulus omission: 0.17 Hz (+/-0.04, 95% CI); second-order stimulus present: 0.13 Hz (+/-0.04, 95% CI)). However, second-order stimuli also changed the pattern of responding whereby rats would make a bout of bar presses prior to the presentation of the second-order stimulus and then pause briefly after the second-order stimulus. In the absence of second-order stimuli, responding was uniformly high. Control measures, such as the ability of the second-order stimulus to evoke checking for the primary reinforcers, indicated that the second-order stimulus was associated with the primary reinforcer.

These results demonstrated that although second-order stimuli maintained responding and caused the rat to check for primary reinforcement, overall response rates were increased when the second-order stimuli were omitted. This has implications for interpreting the results of studies where overall response rates within second-order schedules have been the only measure used to assess the effects of potential anti-addiction drugs. Future studies could be improved by performing a second-order stimulus omission test analysing both the overall response rates and the temporal organization of responding with respect to the second-order stimulus.

Are bCR-ABL transcripts detected in cord blood or the peripheral blood of the newborn child whose mother developed chronic myeloid leukemia while pregnant?

The treatment of choice for the pregnant woman with CML has not been defined. Exposure to imatinib while pregnant may cause serious fetal malformations and interferon-alpha is sometimes associated with side effects. Furthermore, little is known of the possibility that BCR/ABL-positive cells might be passed to the fetus and the role of the treatment given to the pregnant mother. Detection of BCR-ABL transcripts in the peripheral blood of the mother, the newborn and the cord blood was performed by quantitative real time PCR and FISH. A patient with CML diagnosed at the beginning of pregnancy was treated with leukapheresis at 31 weeks of gestation until delivery without any untoward effects. Since no tyrosine kinase inhibitor was administered BCR-ABL transcripts contamination of the cord blood and peripheral blood of the newborn was a reasonable concern. In practice no transcripts were detected in the cord blood or in the peripheral blood of the newborn at birth, at 1 month or 3 at months of age despite the fact that throughout her pregnancy and on the day of delivery the mother had 90% BCR/ABL positive cells in her blood.

Leukapheresis does not eliminate the malignant clone; however the absence of BCR-ABL transcripts in the peripheral blood of the neonate and in the cord blood supports the view that transmission of CML to a fetus is improbable even if the mother's treatment during pregnancy is suboptimal.

Are distinct HIV-1 entry phenotypes associated with transmission , subtype specificity , and resistance to broadly neutralizing antibodies?

The efficiency of CD4/CCR5 mediated HIV-1 entry has important implications for pathogenesis and transmission. The HIV-1 receptor affinity profiling (Affinofile) system analyzes and quantifies the infectivity of HIV-1 envelopes (Envs) across a spectrum of CD4/CCR5 expression levels and distills these data into a set of Affinofile metrics. The Affinofile system has shed light on how differential CD4/CCR5 usage efficiencies contributes to an array of Env phenotypes associated with cellular tropism, viral pathogenesis, and CCR5 inhibitor resistance. To facilitate more rapid, convenient, and robust analysis of HIV-1 entry phenotypes, we engineered a reporter Affinofile system containing a Tat- and Rev-dependent Gaussia luciferase-eGFP-Reporter (GGR) that is compatible with the use of pseudotyped or replication competent viruses with or without a virally encoded reporter gene. This GGR Affinofile system enabled a higher throughput characterization of CD4/CCR5 usage efficiencies associated with differential Env phenotypes. We first validated our GGR Affinofile system on isogenic JR-CSF Env mutants that differ in their affinity for CD4 and/or CCR5. We established that their GGR Affinofile metrics reflected their differential entry phenotypes on primary PBMCs and CD4+ T-cell subsets. We then applied GGR Affinofile profiling to reveal distinct entry phenotypes associated with transmission, subtype specificity, and resistance to broadly neutralizing antibodies (BNAbs). First, we profiled a panel of reference subtype B transmitted/founder (T/F) and chronic Envs (n = 12) by analyzing the infectivity of each Env across 25 distinct combinations of CD4/CCR5 expression levels. Affinofile metrics revealed that at low CCR5 levels, our panel of subtype B T/F Envs was more dependent on high levels of CD4 for HIV-1 entry compared to chronic Envs. Next, we analyzed a reference panel of 28 acute/early subtype A-D Envs, and noted that subtype C Envs could be distinguished from the other subtypes based on their infectivity profiles and relevant Affinofile metrics. Lastly, mutations known to confer resistance to VRC01 or PG6/PG19 BNAbs, when engineered into subtypes A-D Envs, resulted in significantly decreased CD4/CCR5 usage efficiency.

GGR Affinofile profiling reveals pathophysiological phenotypes associated with varying HIV-1 entry efficiencies, and highlight the fitness costs associated with resistance to some broadly neutralizing antibodies.

Is masticatory dysfunction associated with worse functional ability : a population-based study?

Because of the ageing of populations, disability has become an emergent problem from the clinical, social, and economic perspectives. Nevertheless, the determinants of disability in older subjects are still unclear. We assessed the association between self-assessed masticatory dysfunction (MD) and functional ability in older subjects. We analysed data of all 350 subjects aged 75+ living in Tuscania (Italy). Functional ability was estimated using the Katz' activities of daily living (ADLs), and the Lawton and Brody instrumental activities of daily living (IADLs) scales. MD was reported by 145 (41%) participants. Disability in the ADLs and IADLs was found in 37 (25%) and 53 (37%) of participants with MD, respectively, but only in 11 (5%) and 30 (15%) of the other participants (p<0.001). MD was associated with disability in the ADLs [odds ratio (OR)=2.40, 95% confidence interval (CI)=1.05-5.51], and IADLs (OR=2.77, 95% CI=1.07-7.16) in logistic regression, after adjusting. The association of MD with disability was stronger among subjects aged 80+.

MD is independently associated with disability in community-dwelling elderly. Further studies are needed to evaluate the impact of early detection and correction of MD on the preservation of functional status in older populations.

Does high shear stress induce a strain increase in human coronary plaques over a 6-month period?

Atherosclerotic plaques develop in low shear stress regions. In the more advanced phase of the disease, plaques are exposed to altered shear stress levels, which could influence plaque composition. We investigated changes in plaque composition in human coronary arteries over a 6-month period and how these changes are related to shear stress. We took images of eight coronary arteries to obtain the 3D shape of the arteries. Lumen data were combined with computational fluid dynamics to obtain shear stress. Palpography was applied to measure strain at baseline and at 6-month follow-up. The change in strain from baseline to follow-up served as a marker for the change in plaque composition. We identified 17 plaques, and each plaque was divided into four regions: the upstream, throat, shoulder and downstream region. Shear stress and strain in the downstream region was significantly lower than in the other regions. There was no significant change in strain for the four different plaque regions. However, we observed that those plaque regions exposed to high shear stress showed a significant increase in strain.

Plaque regions exposed to high shear stress showed an increase in strain over time. This indicates that shear stress may modulate plaque composition in human coronary arteries.

Is association of juvenile idiopathic arthritis with PTPN22 rs2476601 specific to females in a Greek population?

Juvenile idiopathic arthritis (JIA) is an autoimmune disease characterized by persistent chronic arthritis. Disease risk is believed to be influenced by both genetic and environmental factors. It is well established that the PTPN22 single nucleotide polymorphism (SNP) rs2476601 is associated with JIA susceptibility. It was recently reported in an Australian study that this association is restricted to females and is not observed in males. A significant source of inconsistency amongst the literature on autoimmune disease susceptibility genes stems from an inability to replicate genetic findings across different racial or ethnic groups. We therefore attempted to generate further evidence of the female-specific association of rs2476601 in a homogeneous Greek population. We genotyped rs2476601 in 128 Caucasian JIA patients (70.3 % female) and 221 healthy controls (28.1 % female) from Northern Greece. Overall, PTPN22 was associated with increased risk of JIA in this Greek sample (OR = 2.3, 95 % CI 1.1 - 5.1, p = 0.038). Sex-stratified analyses showed that, once again, the risk association was restricted to females (Female: OR = 19.9, 95 % CI 1.2 - 342, p = 0.0016; Male: OR = 1.1, 95 % CI 0.3 - 3.1, p = 0.94) supporting the prior findings.

Our data demonstrates that this sex-specific pattern of association is broadly applicable to different populations, and provides further impetus to undertake mechanistic studies to understand the impact of sex on PTPN22 in JIA.

Does fas signaling induce raft coalescence that is blocked by cholesterol depletion in human RPE cells undergoing apoptosis?

To investigate whether the signaling events occurring in Fas-mediated apoptosis alter raft membrane formation in human RPE cells. Formation of lipid rafts in cultured human retinal pigment epithelial cells (ARPE-19) was studied by confocal microscopy, with fluorescein-labeled cholera toxin subunit B binding protein (BODIPY)-labeled ganglioside GM1 lipid after Fas-L induction of apoptosis. Apoptosis was assessed by fluorescein-labeled annexin V detection of phosphatidylserine externalization and quadrant analysis with flow cytometry. Membrane rafts were localized into membrane vesicles by passing BODIPY-labeled GM1 RPE cells through a 2-microm-pore polycarbonate membrane using an extruder device. The labeled fractions, containing vesicles enriched in GM1, were detected by flow cytometry and then analyzed for the presence of Fas protein. Differential punctate staining of membrane rafts was demonstrated in normal and FasL-induced apoptotic human ARPE-19 cells in culture by confocal microscopy, using cholera toxin B and GM1 labeling of extruded vesicles. The lipid raft-associated vesicles were derived by plasma membrane dissociation, via a newly developed whole-cell extrusion technique that produced 2-microm vesicles with both GM1 lipid and Fas protein abundance enriched in a subpopulation of the membrane-derived vesicles. The amount of Fas protein in the vesicles containing raft domains markedly increased in FasL-treated cells. Treatment of human ARPE 19 cells with methyl beta-cyclodextrin after FasL induction of apoptosis resulted in cellular cholesterol depletion and markedly reduced the incidence of Fas-receptor localization in GM1 rafts.

Human ARPE-19 cells in culture contain membrane rafts with apoptotic signaling effectors uniformly distributed in the native state. The cells stimulated to undergo apoptosis appear to use membrane rafts in the death-signaling process by mobilization of rafts to localized regions of the membrane that are now enriched with apoptotic signaling effectors. Fas signaling induces apoptotic raft formation that results in polar condensation, or capping, of the rafts in the late stages of apoptosis. A novel extrusion technique is described that allows localization and enrichment of rafts into membrane vesicles, which can be assayed by flow cytometry. Cholesterol depletion, after Fas ligand activation of apoptosis, reduced raft formation in cells induced to undergo apoptosis. Therapeutic implications for the treatment of retinal disorders are discussed.

Does the 'Liverpool Care Pathway' facilitate an improvement in quality of care for dying cancer patients?

The Liverpool Care Pathway for the Dying Patient (LCP) aims to transfer hospice principles of care for dying patients to other health-care sectors. This post-bereavement survey explored the LCP's effectiveness in improving quality of care for cancer patients. Postal self-completion questionnaires were sent to 778 next-of-kin to consecutive deceased patients who had died an 'expected' cancer death in a hospice and acute tertiary hospital. Following exclusions (n=53), 255 of the 725 next-of-kin agreed to participate (35.2% response rate). Overall hospice participants reported the best quality of care, and hospital participants, for whom care was not supported by the LCP, reported the worst quality of care. Multivariate analysis showed the hospice was an independent predictor for patients being treated with dignity (OR 8.46) and receiving adequate family support (OR 7.18) (P<0.0001). Care supported by the LCP and the hospital specialist palliative care team were both associated with good family support, but neither was an independent predictor.

From the bereaved relatives' perspective, within the hospital, the LCP is effective in improving specific aspects of care, such as symptom control for dying patients. Further improvement is required, however, to attain the hospice standard of care.

Is immune response to Bordetella pertussis associated with season and undernutrition in Senegalese children?

While vaccines elicit a protective response in most recipients, studies suggest that environmental and nutritional factors can influence the strength of the individual response to immunization and to subsequent natural infectious challenges. We conducted a longitudinal survey in Senegal to assess the individual response to B. pertussis, a respiratory disease against which Senegalese children are vaccinated before the age of one (Clinicaltrials.gov ID: NCT01545115). A cohort of 203 children aged 1-9 from four villages of the Senegal River Valley was followed-up for 14 months (October 2008-January 2010). During that period, four visits have been made to the villages to assess the immunological and nutritional status of these children and to determine risk factors involved in the modulation of their humoral immune response to B. pertussis toxin. A multivariate model has demonstrated that birth season and nutritional status appeared to modulate humoral response to pertussis toxin. Moreover, response to B. pertussis was dependent on age, village and time of visit.

These results are consistent with the hypothesis that environmental and nutritional factors modulate children's response to pertussis following natural infection or vaccination.

Do pCBs enhance collagen I expression from human peritoneal fibroblasts?

To test the effect of four polychlorinated biphenyl congeners (PCB-77, PCB-105, PCB 153, and PCB 180) on expression of three adhesion markers (transforming growth factor [TGF] beta1, vascular endothelial growth factor [VEGF], and type I collagen) in normal human peritoneal and adhesion fibroblasts. Cell culture study. University research laboratory. Primary cultures of normal peritoneal and adhesion fibroblasts were established from three patients. Fibroblasts were treated with PCB-77, PCB-105, PCB-153, or PCB-180 at 20 ppm for 24 hours. Total RNA was extracted from each treatment and subjected to real-time reverse transcriptase polymerase chain reaction (RT-PCR). MAIN OUTCOME AND MEASURE(S): The mRNA levels of type I collagen, VEGF, and TGF-beta1. Normal human peritoneal fibroblasts expressed type I collagen, VEGF, and TGF-beta1. Exposure of normal human fibroblasts to PCB-77, PCB-105, PCB-153, or PCB-180 did not affect mRNA levels of beta-actin, the housekeeping gene used to normalized RNA levels for the real-time RT-PCR, nor did it affect cell viability as assessed by trypan blue exclusion. The PCB treatments, compared with control, resulted in no significant change for TGF-beta1 or VEGF mRNA levels in normal peritoneal and adhesion fibroblasts. In marked contrast, type I collagen mRNA levels were markedly increased in response to the brief 24 hours' exposure to each PCB treatment in both cell types.

The finding that PCB-77, PCB-105, PCB-153, and PCB-180 increased the expression of type I collagen in human normal peritoneal and adhesion fibroblasts is the first demonstration of involvement of organochlorines in the pathogenesis of tissue fibrosis. This may implicate organochlorine exposure as an etiologic factor in a wide variety of previously unlinked human ailments characterized by fibrosis.

Do bile duct crystals contribute to sphincter of Oddi dysfunction?

Microlithiasis has been proposed as a cause of both occult gallbladder disease and of idiopathic pancreatitis. Theoretically, microlithiasis could also cause postcholecystectomy pain by causing temporary biliary obstruction and may be more common in patients with sphincter of Oddi dysfunction. The frequency of crystals in bile duct aspirates was assessed from patients with symptoms after cholecystectomy with and without elevated baseline sphincter of Oddi pressures. A prospective analysis was performed on all patients with recurrent biliary pain after cholecystectomy who presented for ERCP and manometry between January 1998 and June 2000. All patients had aspirates obtained from the common bile duct for crystal analysis by using the aspirating port of the manometry catheter before the injection of contrast. Four to 20 mL of bile was examined by microscopy for both cholesterol and bilirubinate crystals. Sixty patients (83% women, mean age 44 years) were studied. Thirty-five had normal baseline biliary sphincter pressures and 25 elevated biliary baseline sphincter pressures (>40 mm Hg). Two patients in the normal pressure group and 1 in the elevated pressure group had cholesterol crystals present in their aspirate. No patient had bilirubinate crystals present. A 5% frequency of microlithiasis was identified overall.

Bile duct crystals occur infrequently in patients with symptoms after cholecystectomy and are found in patients with normal and abnormal biliary sphincter manometry. This study suggests that the presence of bile duct crystals, or microlithiasis, does not play a role in sphincter of Oddi dysfunction.

Is polymorphism of 270 A > G in BRAP Associated with Lower Ankle-Brachial Index in a Taiwanese Population?

The single nucleotide polymorphism (SNP) rs11066001 of BRAP has been shown to be associated with myocardial infarction (MI), coronary atherosclerosis and carotid atherosclerosis, but it is not clear whether it also plays a role in peripheral artery disease (PAD). The ankle-brachial index (ABI) is often used as a non-invasive measure of PAD; therefore, the aim of this study was to test for a relationship between the SNP rs11066001 and ABI. A total of 537 high-risk subjects with a family history of MI or stroke completed a health survey, including a physical examination, blood test and measurement of ABI. Among them, 523 subjects had the genotypes. Association analyses between the genotype of BRAP and ABI were performed by multiple linear and logistic regressions with adjustment for covariates. We found that the GG genotype is significantly associated with a lower ABI value. For the lowest ABI tertile, the GG genotype had an OR of 2.87 (p =0.018) when compared with the middle ABI tertile, and OR of 2.92 (p =0.015) when compared to the highest ABI tertile. Women with the GG genotype had a lower ABI value than men with the same genotype (p =0.012). Accordingly, women carrying this GG risk genotype may have a higher risk for PAD.

Our findings provide additional evidence that support the genetic effect of BRAP on diverse cardiovascular phenotypes.

Does upregulation of FasL by LPA on ovarian cancer cell surface lead to apoptosis of activated lymphocytes?

Constitutive expression and upregulation of FasL by malignant epithelial cells counterattack infiltrating natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) and induce apoptosis of normal cells within the tumor, which may induce metastasis. As little is known about the mechanisms that regulate expression of Fas ligand and the subsequent release of FasL in epithelial ovarian cancer cells (EOC), we investigated the effects of lysophosphatidic acid (LPA) on FasL expression and associated signaling pathways. We used established EOC cell lines that were incubated with or without LPA and FasL expression was detected by flow cytometry. Cells were additionally lysed and detected for total protein expression. Activated CD4+ T cells, after coculture with or without EOC, were collected for apoptosis staining and analysis by flow cytometry. Flow cytometry showed that LPA strongly upregulated FasL expression on the OVCAR3 cell surface (P < 0.01), yet in Dov13 cells, LPA significantly upregulated FasL expression only in the presence of the general matrix metalloproteinase (MMP) inhibitors GM6001 and MMP inhibitor II (P < 0.01). The MEK/ERK1/2 kinase cascade is required for FasL upregulation, since the MEK inhibitor PD98059 significantly inhibited FasL upregulation induced by LPA (P < 0.01). Type II secretory phospholipase A2 (sPLA2-II), which promotes protein exocytosis from secretory vesicles and gelatinase granules, affects FasL translocation from intracellular to the cell surface. Pretreatment of Dov13 cells with LPA increased activated T cell apoptosis in cocultures.

These data suggest that upregulation of FasL by LPA provides EOC immune-privilege and leads to apoptosis of activated T lymphocytes.

Does pulsed methylprednisolone induce a reversible impairment of memory in patients with relapsing-remitting multiple sclerosis?

Chronic administration of corticosteroids has been reported to selectively impair explicit memory in systemic diseases without central nervous system involvement. Our aim was to verify that a short course of pulsed intravenous methylprednisolone (IVMP) administered for the treatment of a relapse impairs cognitive functions in relapsing-remitting multiple sclerosis (RRMS) patients and to determine whether this impairment is reversible. Neuropsychological evaluations were made before the start of treatment, and 7 and 60 days after the end of treatment in 14 RRMS patients. The neuropsychological battery was also administered to 12 controls matched for age, sex and years of education. RRMS patients performed worse than the controls at their baseline evaluation for a variety of neuropsychological tasks. IVMP administration induced a selective impairment of explicit memory which completely recovered 60 days after treatment.

In RRMS patients, IVMP induces a selective and reversible impairment of explicit memory.

Are obstructive sleep apnoea and nocturnal gastroesophageal reflux common in lung transplant patients?

Gastroesophageal reflux (GOR) has been implicated in the pathogenesis of bronchiolitis obliterans syndrome (BOS), possibly due to pulmonary aspiration of refluxed acid. Risk of aspiration of gastric contents is increased during sleep due to decreased oesophageal clearance mechanisms and may be further increased by the presence of OSA. This study investigated the relationship between nocturnal GOR, OSA and BOS in a group of lung transplant patients. Fourteen lung transplant patients underwent overnight polysomnography with simultaneous dual oesophageal pH monitoring. Patients had an FEV(1) of 84 +/- 15% of their best post-transplant FEV(1). Six of the 14 patients were in various stages of BOS. The average proportion of time spent overnight with a pH of <4 was 1.7 +/- 3.1%. Increased GOR was evident in 8/14 patients during the postprandial period and/or overnight in the distal and/or proximal oesophagus. All patients had OSA (AHI >5 events per hour). There were no relationships between severity of OSA or GOR and severity of BOS.

Both nocturnal GOR and OSA were common in this group of patients but their occurrences were not related. Neither was there any relationship between the presence of nocturnal GOR or OSA and severity of BOS.

Do caveolae modulate excitation-contraction coupling and beta2-adrenergic signalling in adult rat ventricular myocytes?

Caveolae, flask shaped invaginations of the cell membrane, influence signalling cascades in many cell types. We have tested the hypothesis that caveolae modulate excitation-contraction coupling (ECC) and beta-adrenergic stimulation in the adult cardiac myocyte. Shortening, [Ca(2+)](i) and L-type Ca(2+) current (I(Ca,L)) were recorded in rat ventricular myocytes. Caveolae were disrupted with methyl-beta-cyclodextrin (MbetaC). Shortening and [Ca(2+)](i) transient amplitude were reduced in myocytes treated with MbetaC. MbetaC did not alter the density or characteristics of I(Ca,L) or the sarcoplasmic reticulum (SR) Ca(2+) load, but significantly reduced fractional SR Ca(2+) release. The inotropic response of myocytes to beta(1)-adrenoceptor stimulation was insensitive to caveolae disruption. By contrast, the increase in shortening, [Ca(2+)](i) transient and I(Ca,L) seen following beta(2) stimulation was markedly enhanced (3-5 fold) following MbetaC treatment, and the effect on I(Ca,L) could be mimicked by dialyzing cells with an antibody to caveolin 3. When the G(alphai) pathway was disabled with pertussis toxin (PTX), control cells showed a similar response to beta(2) stimulation as seen in MbetaC-treated myocytes, whereas MbetaC-treated cells were insensitive to PTX.

Caveolae modulate ECC via the efficiency of the Ca(2+)-induced Ca(2+) release process, rather than Ca(2+) influx. Our data are also consistent with the hypothesis that interaction of G(i) protein cascade components with caveolin in the caveolae is necessary for effective signalling by this pathway. This suggests that changes in caveolin expression in the adult heart seen during aging and in disease will have consequences for baseline cardiac function and beta-adrenergic responsiveness.

Evaluation of hands-on training in colonoscopy: is a computer-based simulator useful?

The advantages of using a computer-based simulator during colonoscopy training are debated. We aimed to explore its usefulness in objectively measuring trainees' competence in colonoscopy. Twelve colonoscopy trainees (fully trained in upper GI endoscopy) were evaluated using a computer-based simulator (GI-Mentor, Symbionix) before and during hands-on training (i.e. after 60 colonoscopies); the controls were 15 experts (>90% of caecal intubation). Both trainees and experts performed two "screening" simulations (easy and difficult) in a randomised order, and the time to reach the caecum and withdrawal time was assessed. The percentage of caecal intubation progressively increased during hands-on training. All of the trainees intubated the caecum during the easy and difficult simulations, both before and during hands-on training. The median time (interquartile range) to reach the caecum upon easy simulation was the only variable influenced by hands-on training: 2.7 min (2.1-3.2) before and 1.9 min (1.6-2) during training (p<0.01). Withdrawal time was ≥6 min in the case of five trainees before training, and three during hands-on training. Computer-based simulator performance did not correlate with hands-on training performance.

The computer-based simulator was not found to be useful in evaluating competence during hands-on training in colonoscopy.

Is glutathione-S-transferase P1 , T1 and M1 genetic polymorphisms in neoadjuvant-treated locally advanced gastric cancer : GSTM1-present genotype associated with better prognosis in completely resected patients?

Neoadjuvant chemotherapy in gastric cancer is now standard in the Western world; however, only 30-40% of the patients respond to induction therapy. Pretherapeutic predictors of response and prognosis would be of utmost interest to individualize treatment. Glutathione-S-transferase enzymes detoxify therapeutic drugs such as platin derivates and may influence outcome of the treated patients. Therefore, glutathione-S-transferase (GST) polymorphisms were assessed as predictive markers in cisplatinum-based neoadjuvant-treated gastric cancer. DNA was isolated from 139 patients with locally advanced gastric cancer (cT3/4 anyN cM0) before chemotherapy. Multiplex polymerase chain reaction was used for GSTT1 and GSTM1 genes, and allelic discrimination assay with the TaqMan system for the GSTP1 gene. One hundred ten patients could be analyzed for GSTT1 (T-:23; T+87), 112 for GSTM1 (M-:52; M+:60) and 132 for GSTP1 (Ile/Ile: 55; Ile/Val: 59; Val/Val: 18). There was no significant correlation between any of the GSTT1, GSTM1, or GSTP1 genotypes and patients' characteristics or histopathological data; only the GSTM1+ genotype was associated with the non-intestinal subtype of the Lauren classification (p=0.045). GSTT1, GSTM1, and GSTP1 genotypes were not correlated with response to chemotherapy (p=0.57, p=0.38, p=0.33). In R0 resected patients, we found an improved survival for patients with the GSTM1-present genotype compared to patients with the GSTM1-null genotype (p=0.017). Moreover, the GSTM1-present genotype showed a significantly better tumor-related (p=0.017) and disease-free survival (p=0.029).

None of the common GST polymorphisms predicts response in our study, but the GSTM1+ genotype was associated with a better prognosis in completely resected patients. Further investigations on chemotherapy-associated gene polymorphisms are warranted.

Is fiberoptic monitoring of central venous oxygen saturation ( PediaSat ) in small children undergoing cardiac surgery : continuous continuous?

 Monitoring of superior vena cava saturation (ScvO 2) has become routine in the management of pediatric patients undergoing cardiac surgery. The objective of our study was to evaluate the correlation between continuous ScvO 2 by the application of a fiber-optic oximetry catheter (PediaSat) and intermittent ScvO 2 by using standard blood gas measurements. These results were compared to those obtained by cerebral near infrared spectroscopy (cNIRS).  Tertiary pediatric cardiac intensive care unit (PCICU).  A retrospective study was conducted in consecutive patients who were monitored with a 4.5 or 5.5 F PediaSat catheter into the right internal jugular vein. An  in vivo calibration was performed once the patient was transferred to the PCICU and re-calibration took place every 24 hours thereafter. Each patient had a NIRS placed on the forehead. Saturations were collected every 4 hours until extubation. Ten patients with a median age of 2.2 (0.13-8.5) years and a weight of 12.4 (3.9-24) kg were enrolled. Median sampling time was 32 (19-44) hours: 64 pairs of PediaSat and ScVO2 saturations showed a poor correlation (r=0.62, 95% CI 44-75; p<0.0001) and Bland Altman analysis for repeated measures showed an average difference of 0.34 with a standard deviation of 7,9 and 95% limits of agreement from -15 to 16. Thirty-six pairs of cNIRS and ScVO2 saturations showed a fair correlation (r=0.79, 95% CI 0.60-0.89; p<0.0001) an average difference of -1.4 with a standard deviation of 6 and 95% limits of agreement from -13 to 10. Analysis of median percentage differences between PediaSat and ScvO2 saturation over time revealed that, although not statistically significant, the change in percentage saturation differences was clinically relevant after the 8th hour from calibration (from -100 to +100%).

 PediaSat catheters showed unreliable performance in our cohort. It should be further investigated whether repeating calibrations every 8 hours may improve the accuracy of this system. CNIRS may provide similar results with a lower invasiveness.

Does angiopoietin-1 inhibit irradiation- and mannitol-induced apoptosis in endothelial cells?

Angiopoietin-1 (Ang1) is a vasculogenic factor that signals through the endothelial cell-specific Tie2 receptor tyrosine kinase. We recently reported that Ang1 prevented apoptosis induced by serum deprivation in endothelial cells. In this study, we examined whether Ang1 prevents apoptosis in endothelial cells treated with irradiation or clinical concentrations of mannitol. ++Ang1 prevented irradiation- and mannitol-induced apoptosis in human umbilical vein endothelial cells in a dose-dependent manner. Pretreatment with soluble Tie2 receptor, but not Tie1 receptor, blocked the antiapoptotic effect of Ang1. Two phosphatidylinositol 3'-kinase (PI3-kinase)-specific inhibitors, wortmannin and LY294002, blocked the Ang1-induced antiapoptotic effect. The antiapoptotic potency of Ang1 was similar to or greater than that of vascular endothelial growth factor, basic fibroblast growth factor, and endothelin-1. Ang1 also prevented apoptosis in cultured endothelial cells from porcine pulmonary and coronary arteries and in endothelial cells of explanted rat aorta.

Ang1 promotes the survival of endothelial cells in irradiation- and mannitol-induced apoptosis through Tie2 receptor binding and PI3-kinase activation. Pretreatment with Ang1 could be beneficial in maintaining normal endothelial cell integrity during intracoronary irradiation or systemic mannitol therapy.

Does ileo-anal pouch surgery in a district general hospital result in good outcomes and high patient satisfaction?

Restorative proctocolectomy (RPC) with ileal-pouch anal anastomosis (IPAA) is a technically challenging procedure. This study aims to review the outcomes following surgery carried out in a DGH by two surgeons with experience gained in tertiary centres. All patients undergoing RPC with IPAA were identified from a prospectively collected database and case notes reviewed. Data were collected on demographics, indication for surgery, operative details, outcomes and adherence to a treatment and follow-up protocol developed with a specialist centre. A validated questionnaire (the Pouch Functional Score) was sent to patients to assess functional outcome. Fourteen patients (nine male) underwent surgery from 2008 to 2012, average age 32 years (range 22-48). Median follow up was 5 years (interquartile range 3.2 years). The indication for surgery was ulcerative colitis. All patients had a 'J' shaped pouch, stapled anastomosis and defunctioning ileostomy. There was no operative mortality. One patient was re-admitted with high ileostomy output; three developed wound infections. All patients have had their stomas closed. There were no postoperative cases of pelvic sepsis or anastomotic leak. The median stool frequency is 6-8/24hrs, 23% of patients reported urgency, 23% had occasional incontinence. The protocol was adhered to in all cases. One patient required defunctioning of the pouch 5 years after surgery.

With good patient selection, a team approach and a protocol ensuring consistent care, pouch surgery is being performed to a high standard in a DGH setting. Follow-up care is provided locally and patients have easy access to the multidisciplinary team.

Is posttreatment platelet reactivity on clopidogrel associated with the risk of adverse events after off-pump coronary artery bypass?

Dual antiplatelet therapy with aspirin and clopidogrel is currently recommended in off-pump coronary artery bypass (OPCAB). However, no data exist concerning platelet reactivity on clopidogrel after OPCAB. The aim of this study was to assess the relationship between platelet reactivity and late major adverse cardiovascular events (MACEs) after OPCAB. In this prospective, single-center, observational study, on-clopidogrel platelet reactivity was measured using a point-of-care assay (VerifyNow system; Accumetrics Inc, San Diego, CA) in 859 patients who underwent OPCAB with 1 or more vein grafts. The primary end point was late MACEs (30 days-1 year) including cardiac death, nonfatal myocardial infarction, and target vessel revascularization. Receiver operating characteristic curve analysis was used to estimate the cutoff value of P2Y12 reaction units (PRUs) for MACEs. The optimal cutoff value for posttreatment reactivity for the incidence of late MACEs was ≥188 PRU (area under the curve 0.72, 95% CI 0.68-0.75, P = .002). The incidence of late MACEs was significantly higher in the high platelet reactivity (HPR; ≥188 PRU) group than in the low platelet reactivity (<188 PRU) group (3.6% vs. 1.4%, P = .040). Kaplan-Meier analysis revealed 1-year MACE-free survival rates of 98.4% ± 0.5% and 95.9% ± 1.3% in the low platelet reactivity and HPR groups, respectively (P = .034). According to a Cox regression hazard model, HPR was an independent risk factor for late MACE-free survival (hazard ratio 3.51, 95% CI 1.27-9.69, P = .015).

High residual platelet reactivity after clopidogrel administration is strongly associated with 1-year MACE-free survival. Routine measurement of platelet reactivity and thorough monitoring of patients with HPR after OPCAB are warranted.

Does methadone maintenance treatment based on the new national guidelines work in a primary care setting?

General practitioners (GPs) are being encouraged to treat more drug users but there are few studies to demonstrate the effectiveness of primary care treatment.AIM: To determine whether patients retained on methadone maintenance treatment for one year in a modern British primary care setting, with prescribing protocols based on the new national guidelines, can achieve similar harm reduction outcomes to those demonstrated in other settings, using objective outcome measures where available. Longitudinal cohort study. The Primary Care Clinic for Drug Dependence, Sheffield. The intervention consisted of a methadone maintenance treatment provided by GPs with prescribing protocols based on the 1999 national guidelines. The first 96 eligible consenting patients entering treatment were recruited; 65 completed the study. Outcome measures were current drug use, HIV risk-taking behaviour, social functioning, criminal activity, and mental and physical health, supplemented by urinalysis and criminal record data. Frequency of heroin use was reduced from a mean of 3.02 episodes per day (standard deviation [SD] = 1.73) to a mean of 0.22 episodes per day (SD = 0.54), (chi 2 = 79.48, degrees of freedom [df]= 2, P<0.001), confirmed by urinalysis. Mean numbers of convictions and cautions were reduced by 62% (z = 3.378, P<0.001) for all crime. HIV risk-taking behaviour, social functioning, and physical and psychological wellbeing all showed significant improvements.

Patients retained on methadone maintenance treatment for one year in a primary care setting can achieve improvements on a range of harm reduction outcomes similar to those shown by studies in other, often more highly structured programmes.

Does the broad autism phenotype predict child functioning in autism spectrum disorders?

Broad autism phenotype (BAP) is a milder expression of the social and communication impairments seen in autism spectrum disorders (ASD). While prior studies characterized the BAP in unaffected family members of probands with ASD, the relationship between parental BAP traits and proband symptomatology remains poorly understood. This study utilizes the Broad Autism Phenotype Questionnaire (BAPQ) in parents and the Social Responsiveness Scale (SRS) in children to examine this connection. We hypothesized that in families affected by ASD, elevated maternal and paternal BAPQ scores would correlate with greater autism symptomatology in diagnosed children. In an extension of prior research, we also explored this relationship in families with typically developing children (TDC). Two hundred and forty-five children with ASD, 129 TDC and all parents were recruited as part of a larger study investigating relationships between genes, brain and behavior. The Autism Diagnostic Interview-Revised (ADI-R), Autism Diagnostic Observation Schedule (ADOS) and expert clinical judgment confirmed ASD diagnoses in children. SRS was collected for all children. Parents completed a self-report BAPQ and an informant report BAPQ for their spouse; an average of self-report and informant report for each parent was used in all analyses. Mothers and fathers of children with ASD had significantly higher rates of BAP traits as compared to parents of TDC. Maternal and paternal BAPQ total scores were not correlated with child IQ in either group. In the ASD group, 10% of mothers and 21% of fathers scored above the established BAP threshold compared to 4% of TDC parents. Crude regression analyses showed that maternal and paternal BAPQ total scores accounted for significant variance in child SRS scores in both ASD (17.1%) and TDC (19.8%) families.

Our results suggest that broad autism symptomatology in parents is moderately associated with their child's autism symptomatology. This result extended to TDC families, suggesting that the BAPQ and SRS capture subtle, subclinical social variation in both children and adults. These findings could help define multi-generational social impairments in future phenotypic and genetic studies.

Is referent d-dimer enzyme-linked immunosorbent assay testing of limited value in the exclusion of thromboembolic disease : result of a practical study in an ED?

The aim of this study was to assess in clinical practice the accuracy of a referent d-dimer enzyme-linked immunosorbent assay for the exclusion of venous thromboembolic disease (VTED). An observational prospective study took place in an emergency department; 205 consecutive outpatients suspected of having VTED were included. Blood samples were collected at admission for VIDAS DD measurement. Venous thromboembolic disease was confirmed by standard clinical imaging. All patients were followed up at 3 months. Venous thromboembolic disease was confirmed in 57 patients (28%). The sensitivity and negative predictive value of a DD assay lower than 500 ng/mL were 78% (95% confidence interval = 67%-87%) and 84% (95% confidence interval = 73%-90%), respectively. Twelve patients had a false-negative DD with one or more of the following: (a) symptoms reported for more than 15 days (n = 2), (b) prior anticoagulation (n = 3), (c) distal VTED (n = 5), or (d) high clinical probability (n = 3).

In our cohort of patients, DD was less accurate than previously reported, with an upper estimate of the sensitivity of only 87%.

Is β-cell dysfunction in women with previous gestational diabetes associated with visceral adipose tissue distribution?

Glucose intolerance in pregnancy predicts an increased risk of future type 2 diabetes. The aim of the study was to evaluate glucose metabolism in women with and without gestational diabetes mellitus (GDM) at 5 years follow-up and identify risk factors associated with disturbed glucose metabolism post-partum. This follow-up study included 300 consecutively enrolled women from a previous population-based cohort study. The participants underwent oral glucose tolerance test under pregnancy and in the follow-up study, in addition to dual-energy X-ray absorptiometry in the follow-up study. Fifty-two women (17.7%) were found to have GDM in pregnancy with an odds ratio of 4.8 developing prediabetes 5 years later. β-cell function, but not insulin resistance or sensitivity, was reduced in the follow-up study after adjusting for known risk factors. Furthermore, visceral fat content at follow-up was increased in GDM women compared to non-GDM women, and the β-cell function declined with increasing visceral fat in both groups but was more pronounced in the women with previous GDM.

Women with GDM are at increased risk of developing prediabetes and have a decreased β-cell function 5 years post-partum that is associated with increased visceral fat mass.

Does arecoline inhibit catecholamine release from perfused rat adrenal gland?

To study the effect of arecoline, an alkaloid isolated from Areca catechu, on the secretion of catecholamines (CA) evoked by cholinergic agonists and the membrane depolarizer from isolated perfused rat adrenal gland. Adrenal glands were isolated from male Sprague-Dawley rats. The adrenal glands were perfused with Krebs bicarbonate solution by means of a peristaltic pump. The CA content of the perfusate was measured directly using the fluorometric method. Arecoline (0.1-1.0 mmol/L) perfused into an adrenal vein for 60 min produced dose- and time-dependent inhibition in CA secretory responses evoked by acetylcholine (ACh) (5.32 mmol/L), 1.1-dimethyl-4-phenyl piperazinium iodide (DMPP) (100 micromol/L for 2 min) and 3-(m-choloro-phenyl-carbamoyl-oxy)-2-butynyl trimethyl ammonium chloride (McN-A-343) (100 micromol/L for 2 min). However, lower doses of arecoline did not affect CA secretion of high K(+) (56 mmol/L); higher doses greatly reduced CA secretion of high K(+). Arecoline also failed to affect basal catecholamine output. Furthermore, in adrenal glands loaded with arecoline (0.3 mmol/L), CA secretory response evoked by Bay-K-8644 (10 micromol/L), an activator of L-type Ca(2+) channels, was markedly inhibited, whereas CA secretion by cyclopiazonic acid (10 micromol/L), an inhibitor of cytoplasmic Ca(2+)-ATPase, was not affected. Nicotine (30 micromol/L), which was perfused into the adrenal gland for 60 min, however, initially enhanced ACh-evoked CA secretory responses. As time elapsed, these responses became more inhibited, whereas the initially enhanced high K(+)-evoked CA release diminished. CA secretion evoked by DMPP and McN-A-343 was significantly depressed in the presence of nicotine.

Arecoline dose-dependently inhibits CA secretion from isolated perfused rat adrenal gland evoked by activation of cholinergic receptors. At lower doses arecoline does not inhibit CA secretion through membrane depolarization, but at larger doses it does. This inhibitory effect of arecoline may be mediated by blocking the calcium influx into the rat adrenal medullary chromaffin cells without the inhibition of Ca(2+) release from the cytoplasmic calcium store. There seems to be a difference in the mode of action of nicotine and arecoline in rat adrenomedullary CA secretion.

Does chronic intermittent hybobaric hypoxia protect against cerebral ischemia via modulation of?

Providing adequate protection against cerebral ischemia remains an unrealized goal. The present study was aimed at testing whether chronic intermittent hypobaric hypoxia (CIHH) would have protective effects against cerebral ischemia and investigating the potential role of mitochondrial membrane ATP-sensitive potassium channel (mitoK Ischemia was induced in rats by occlusion of bilateral common carotid arteries for 8min on day 2 after bilateral vertebral arteries were permanently electrocauterized and CIHH was simulated in a hypoxic chamber. Learning and memory impairments were analyzed using the Morris water maze. The delay neuronal death (DND) in the hippocampus CA1 was observed by thionine staining. The expression of the two subunits of mitoK CIHH pretreatment ameliorated the learning and memory impairments produced by ischemia, concomitant with reduced DND in the hippocampus CA1 area. Expression levels of SUR1 and Kir6.2 both increased for at least one week after CIHH pretreatment. Levels of the two subunits were higher in the CIHH pretreatment combined with ischemia group than the ischemia only group at 2 d and 7 d after ischemia. Furthermore, the concentration of Cyt c was decreased in mitochondria and increased in the cytoplasm after ischemia which was prevented by CIHH. The decrease of Δψm and the destruction of mitochondrial ultrastructure were both rescued by CIHH pretreatment. The above protective effects of CIHH were blocked by 5-HD intraperitoneal injection 30min before ischemia.

CIHH pretreatment can reduce cerebral ischemic injury, which is mediated by upregulating the expression and activity of mitoK

Does replacement of the proximal aorta add no further risk to aortic valve procedures?

Aortic valve pathology is often associated with proximal aortic dilatation. Even after valve surgery, the proximal aorta can continue to dilate and thus be at risk for rupture, dissection, or later aortic replacement. We hypothesized that the addition of proximal aortic intervention adds no further risk to aortic valve surgery, which may avoid subsequent proximal aortic procedures or catastrophes. Between 1996 and 2004, 430 aortic valve interventions alone and 146 aortic valves with proximal aortic replacements were identified in elective adult patients. The age in the valve-alone patients (68.8 years) was slightly higher than the valve-plus-aorta group (valve/aorta, 60.5 years; p < 0.01), but comorbidities were similar between groups. We compared groups based on hospital mortality and incidence of complications. The 30-day mortality was similar between groups (valve-alone, 3.8% versus valve/aorta, 2.7%; p = 0.5), as were rates for bleeding and operative revision (valve-alone, 6.7% versus valve/aorta, 9.5%; p = 0.5). Pulmonary (valve-alone, 23.0% versus valve/aorta, 11.6%) and renal complications (valve-alone, 8.2% versus valve/aorta, 2.7%) were higher in the valve-alone group (p = 0.02). Logistic regression demonstrated no additional risk of death, neurologic, or cardiac event with replacement of the proximal aorta.

Proximal aortic replacement adds no risk to the patient beyond the aortic valve intervention alone. These findings suggest proximal aortic replacement is safe for patients undergoing valve operations. Patients with a moderately enlarged proximal aorta that may dilate further should also be considered for aortic replacement at the time of valve procedures.

Does gamma-secretase inhibitor reduce allergic pulmonary inflammation by modulating Th1 and Th2 responses?

Gamma-secretase inhibitor (GSI) has been used to effectively block Notch signaling, which is implicated in the differentiation and functional regulation of T helper (Th) effector cells. In asthma, a subset of CD4(+) T cells is believed to initiate and perpetuate the disease. The aim of this study was to evaluate the therapeutic potential of GSI against allergic asthma. GSI was administered to an ovalbumin-sensitized mouse via an intranasal route at the time of ovalbumin challenge. The administration of GSI inhibits asthma phenotypes, including eosinophilic airway inflammation, goblet cell metaplasia, methacholine-induced airway hyperresponsiveness, and serum IgE production. GSI treatment of bronchoalveolar lavage cells stimulated via TCR or non-TCR pathways led to a decrease in Th2 cytokine production with a concomitant increase in Th1 cytokine secretion. Expression of Hes-1, a target of Notch signaling, was down-regulated in conjunction with a reduction of Notch intracellular domain and GATA-3 levels after GSI treatment of bronchoalveolar lavage cells. GSI treatment resulted in an inhibition of NF-kappaB activation, and combined treatment with GSI and an NF-kappaB inhibitor augmented IFN-gamma production in a synergistic manner.

These data suggest that GSI directly regulates Th1 and Th2 responses in allergic pulmonary inflammation through a Notch signaling-dependent pathway and that GSI is of high therapeutic value for treating asthma by inhibiting airway inflammatory responses.

Does sterilization of platelet concentrate at production scale by irradiation with short-wave ultraviolet light?

Bacterial contamination of platelet concentrates (PCs) is recognized as a serious threat to transfusion safety. We developed a simple method for sterilization of PCs with short-wave ultraviolet light (UVC). The effects of treatment on the sterility of contaminated PCs and in vitro platelet (PLT) variables were evaluated. Plasma-reduced PCs were prepared from pools of five buffy coats. Irradiation with UVC (wavelength, 254 nm) under vigorous agitation was from both sides of the irradiation bags. Kinetics of the inactivation of Bacillus cereus, Propionibacterium acnes, and Staphylococcus epidermidis were determined. PCs spiked with approximately 10 to 100 colony-forming units (CFUs)/mL of 10 bacteria species (n = 12/species) were irradiated with UVC doses between 0.25 and 0.4 J/cm(2) and tested for sterility by a commercially available bacterial detection system (BacT/ALERT, bioMérieux) after storage at 22 degrees C for 3 or 6 days. The influence of a dose of 0.3 J/cm(2) on PLT variables was investigated on Days 1, 4, and 6 after irradiation. At 0.3 J/cm(2) all bacteria species tested were inactivated by more than 4 log. At this dose the influence of UVC on in vitro PLT variables was marginal; the storage stability for up to 6 days after treatment was maintained. PCs spiked with approximately 10 to 100 CFUs/mL were reproducibly sterilized in the dose range tested. In individual experiments with the spore former B. cereus, PCs were, however, unsterile after treatment.

Irradiation at UVC doses not detrimental to in vitro PLT variables sterilizes PCs contaminated with a wide range of different bacteria species.

Do why blacks take part in HIV vaccine trials?

AIDS is still a major cause of death. To combat this disease, researchers are developing a vaccine. Although blacks account for most new infections in the United States, they account for a low percent of experimental vaccine recipients. This study, conducted in a mid-sized U.S. city where vaccine trials are held, seeks to learn why. We conducted 11 in-depth ethnographic interviews. Two groups were targeted: blacks who had not participated in HIV vaccine trials and blacks who had. Overall, three major causes of nonparticipation were identified: misinformation, fear/mistrust and stigma. Factors that favored participation included having close friends with HIV and being homosexual.

HIV is considered by many blacks to be a gay, white disease. Steps to increase participation must include efforts to destigmatize the condition and disseminate accurate information. Efforts to address historical causes of mistrust through "education" alone are insufficient. Trust needs to be earned through long-term relationships with black communities.

Warm water or oil-assisted colonoscopy: toward simpler examinations?

Completion rates, pain, and difficulties during the exam are still problems in colonoscopy. New methods of lubrication, rarely considered a matter of study, may help in this respect. Our aim was to compare an oil-assisted technique with a modified warm water method applied during colonoscopy. A prospective, randomized, and controlled study was planned in which three groups of patients were submitted to colonoscopy: a standard lubricating method (water-soluble jelly: group A, 170 patients) was adopted in a control group, whereas the standard method plus injection into the colon of corn seed oil (group B, 170 patients) or warm water (group C, 170 patients) were employed in the other groups. The main variables evaluated were: the success rate for total intubation, the time required to reach the cecum and the time needed to examine the colon at withdrawal, and the level of pain and degree of difficulty associated with the examination. Successful intubation to the cecum was significantly more frequent (P<0.01 and P<0.001, respectively) in the oil group (group B, 155/166) and in the warm water group (group C, 156/163) than in the control group (group A, 138/164), and less time was needed (P<0.001); no significant difference was found between group B and C. Furthermore, no significant differences were found with regard to time for examination at withdrawal among the three groups. Level of pain and degree of difficulty during colonoscopy were significantly lower in the oil (P<0.001) and in the warm water (P<0.001) groups than in the control group, but no significant difference was found between group B and C. Neither side effects were observed for patients nor damage to the instrument.

Warm water and oil-assisted colonoscopy could be simple, safe, and inexpensive methods for easier and less painful examinations.

Is extensively necrotic retinoblastoma associated with high-risk prognostic factors?

Retinoblastoma is the most common malignant intraocular tumor in children. It has been shown that adjuvant therapy following enucleation in patients with high-risk histopathologic features significantly decreases the mortality. We describe the association of extensive necrosis of tumor and intraocular structures with 2 of the major risk factors: optic nerve invasion and choroidal invasion. This may alert the pathologist who makes the observation of extensive necrosis to carefully search for histologic features associated with adverse outcome. To determine whether extensively necrotic retinoblastoma is associated with high-risk histologic prognostic factors for metastatic disease and patient survival. Retrospective case series. Forty-three eyes of 43 patients with retinoblastoma who underwent enucleation between 1990 and 2001 were evaluated. Medical records, histopathology specimens, pathology reports, and clinical photographs were reviewed. Tumors were designated as exhibiting extensive necrosis if more than 95% of tumor cells and intraocular tissues were necrotic. The main outcome measure was the association of extensive tumor necrosis with 3 high-risk histopathologic features: extraocular extension, optic nerve invasion, or choroidal invasion. Metastatic disease, patient survival, and associations with pathologic findings were also analyzed. Optic nerve head invasion (P < .001), post-lamina-cribrosal invasion (P < .001), and choroidal invasion by tumor (P = .004) were observed more frequently in eyes with extensive necrosis compared with eyes without extensive necrosis. Two of the 11 patients with extensively necrotic intraocular retinoblastoma died from metastatic disease (P = .06). None of the 32 patients without extensive necrosis developed metastatic disease or died.

Extensive ocular tissue and tumor necrosis is associated with histologic high-risk prognostic factors for tumor metastasis and mortality.

Does constitutively active adenosine monophosphate-activated protein kinase regulate voltage-gated sodium channels in ventricular myocytes?

Some PRKAG2 mutations in the human gene encoding for the gamma-subunit of the adenosine monophosphate-activated protein kinase (AMPK) recently have been shown to cause rhythm disturbances (often fatal) in affected patients. Rat ventricular myocytes were infected with an adenoviral vector designed to express a truncated constitutively active mutant (T172D) of the AMPK alpha1-subunit (CA-AMPK). The human cardiac sodium channel hH1 and CA-AMPK were also coexpressed in a mammalian cell line. Patch-clamp techniques were used to measure myocyte action potentials and recombinant hH1 sodium channel currents. Our results demonstrate that action potential duration is significantly prolonged in myocytes expressing the CA-AMPK construct, leading to the production of potentially arrhythmogenic early afterdepolarizations. Recombinant sodium channel current analysis revealed that expression of CA-AMPK significantly slowed open-state inactivation and shifted the voltage-activation curve in a hyperpolarizing direction.

We propose that sodium channels may be substrates for AMPK, possibly contributing to the observed arrhythmogenic activity in patients with some PRKAG2 mutations.

Does in Silico identification of pathogenic strains of Cronobacter from Biochemical data reveal association of inositol fermentation with pathogenicity?

Cronobacter, formerly known as Enterobacter sakazakii, is a food-borne pathogen known to cause neonatal meningitis, septicaemia and death. Current diagnostic tests for identification of Cronobacter do not differentiate between species, necessitating time consuming 16S rDNA gene sequencing or multilocus sequence typing (MLST). The organism is ubiquitous, being found in the environment and in a wide range of foods, although there is variation in pathogenicity between Cronobacter isolates and between species. Therefore to be able to differentiate between the pathogenic and non-pathogenic strains is of interest to the food industry and regulators. Here we report the use of Expectation Maximization clustering to categorise 98 strains of Cronobacter as pathogenic or non-pathogenic based on biochemical test results from standard diagnostic test kits. Pathogenicity of a strain was postulated on the basis of either pathogenic symptoms associated with strain source or corresponding MLST sequence types, allowing the clusters to be labelled as containing either pathogenic or non-pathogenic strains. The resulting clusters gave good differentiation of strains into pathogenic and non-pathogenic groups, corresponding well to isolate source and MLST sequence type. The results also revealed a potential association between pathogenicity and inositol fermentation. An investigation of the genomes of Cronobacter sakazakii and C. turicensis revealed the gene for inositol monophosphatase is associated with putative virulence factors in pathogenic strains of Cronobacter.

We demonstrated a computational approach allowing existing diagnostic kits to be used to identify pathogenic strains of Cronobacter. The resulting clusters correlated well with MLST sequence types and revealed new information about the pathogenicity of Cronobacter species.

Does wnt5a promote Cytokines Production and Cell Proliferation in Human Hepatic Stellate Cells Independent of Canonical Wnt Pathway?

Wnt5a is involved in the activation of human hepatic stellate cells (HSC) and related with the occurrence of liver fibrosis. As the function and mechanism that Wnt5a mediates HSC activation remains unclear, we sought to investigate them. Wnt5a levels were determined in the HSC cell line LX-2 after lipopolysaccharide (LPS) and TNF-α stimulation. HSC cells showing stable and efficient overexpression or featuring knockdown of Wnt5a were constructed by a lentivirus system. Regulation of cytokine and collagen expressions were confirmed by quantitative PCR or ELISA in stable LX-2 cell lines showing Wnt5a overexpression or knockdown. Proliferation was determined by 5-ethynyl-2'-deoxyuridine labeling. Relevant signaling pathways were identified using specific protein antibodies. LPS and TNF-α induced Wnt5a expression in LX-2 cells. Compared with control cells, an increase in IL-10, IL-6, COL1, and COL3 secretion in a stable LX-2 cell line showing Wnt5a overexpression was observed. Knockdown of Wnt5a obviously reduced the production of IL-1β, IL-6, COL1, and COL3. Wnt5a overexpression promoted LX-2 proliferation, while Wnt5a knockdown dramatically inhibited cell proliferation. Compared with the effects of Wnt5a knockdown cells, Wnt5a-overexpressing cells triggered the phosphorylation of c-Jun N-terminal kinase (JNK) and β-catenin, which leads to β-catenin degradation and inactivates the canonical Wnt/β-catenin pathway.

Wnt5a regulates inflammatory cytokine and collagen production and cell proliferation, which is independent of the canonical Wnt signaling pathway. The results reveal a new signaling mechanism in HSC activation and could provide a new strategy for hepatic fibrosis treatment.

Does tNFα suppress IFNγ-induced MHC class II expression on retinal pigmented epithelial cells cultures?

One major consequence of retinal pigment epithelium (RPE) cell activation during autoimmune uveitis is the induction of MHC II molecules expression at their surface. IFNγ is regarded as the main cytokine involved in this induction. As TNFα plays a central role in autoimmune uveitis, we investigated its effects on IFNγ-mediated MHC II induction on RPE cells. Retinal pigment epithelium cells (ARPE-19) were stimulated with IFNγ, TNFα and the anti-TNFα antibody infliximab. The expression of MHCII and ICAM-1 was analysed by flow cytometry. The activation and expression of IRF-1 and STAT-1, two proteins involved in IFNγ-signalling pathway, were analysed by WB. Class II transactivator (CIITA) expression was monitored by qRT-PCR and immunoprecipitation. TNFα inhibits IFNγ-induced MHC II expression on ARPE cells in a dose-dependent manner. Infliximab completely reverses the inhibitory effect of TNFα. We did not observe an inhibitory effect of TNFα on the expression of ICAM-1 induced by IFNγ. Similarly, IFNγ-induced STAT1 phosphorylation and IRF1 expression were not affected by TNFα. On the contrary, we found that TNFα suppresses IFNγ-induced CIITA mRNA accumulation and protein expression.

TNFα inhibits IFNγ-induced MHC II expression in RPE cells. This inhibitory effect was reversed by infliximab and was not because of a global inhibition of IFNγ -mediated RPE cell activation but rather to a specific down-regulation of CIITA expression. Those findings are consistent with the role of TNFα in the resolution of inflammation and might help to elucidate the complex development of autoimmune uveitis.