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Does localization of pacing and defibrillator lead using standard x-ray views is frequently inaccurate and is not reproducible?

While the accuracy of roentography for evaluation of lead tip position compared with three-dimensional imaging techniques has been well described, there remains considerable variability in the interpretation of the reproducibility of standard x-ray for right ventricular (RV) and left ventricular (LV) lead position. The aim of this study was to evaluate the accuracy and reliability of right ventricular (RV) and left ventricular (LV) lead tip position as determined by board-certified cardiac electrophysiologists (EP) using standard x-ray views. EP interpretations of RV and LV lead tip position using standard x-ray views (posterior-anterior, lateral, and left anterior oblique) were compared to thoracic computed tomography (CT). The accuracy of x-ray interpretation was compared to the reference CT location, and the reproducibility of x-ray interpretation was tested using the free-marginal Kappa statistic. A total of 58 EPs were invited to participate in the survey with a response rate of 43 % (25/58). The agreement between x-ray and CT lead tip position (accuracy) was 37 % for RV lead, 33 % for longitudinal LV lead, and 41 % for short axis LV lead. Reproducibility was 64 % for RV lead tip (k = 0.46), 58 % for longitudinal LV lead tip (k = 0.37), and 39 % for short axis LV lead tip (k = 0.24).

Conventional roentography is limited in its ability to accurately and reliably determine pacing lead tip position.

Do retractile testes have anatomical anomalies?

To assess the incidence of anatomical anomalies in patients with retractile testis. We studied prospectively 20 patients (28 testes) with truly retractile testis and compared them with 25 human fetuses (50 testes) with testis in scrotal position. We analyzed the relations among the testis, epididymis and patency of the processus vaginalis (PV). To analyze the relations between the testis and epididymis, we used a previous classification according to epididymis attachment to the testis and the presence of epididymis atresia. To analyze the structure of the PV, we considered two situations: obliteration of the PV and patency of the PV. We used the Chi-square test for contingency analysis of the populations under study (p<0.05). The fetuses ranged in age from 26 to 35 weeks post-conception (WPC) and the 20 patients with retractile testis ranged in ages from 1 to 12 years (average of 5.8). Of the 50 fetal testes, we observed complete patency of the PV in 2 cases (4%) and epididymal anomalies (EAs) in 1 testis (2%). Of the 28 retractile testes, we observed patency of the PV in 6 cases (21.4%) and EA in 4 (14.28%). When we compared the incidence of EAs and PV patency we observed a significantly higher prevalence of these anomalies in retractile testes (p=0.0116).

Retractile testis is not a normal variant with a significant risk of patent processos vaginalis and epididymal anomalies.

Is childhood ADHD strongly associated with a broad range of psychiatric disorders during adolescence : a population-based birth cohort study?

To evaluate associations between attention-deficit/hyperactivity disorder (ADHD) and comorbid psychiatric disorders using research-identified incident cases of ADHD and population-based controls. Subjects included a birth cohort of all children born 1976-1982 remaining in Rochester, MN after age five (n = 5,718). Among them we identified 379 ADHD incident cases and 758 age-gender matched non-ADHD controls, passively followed to age 19 years. All psychiatric diagnoses were identified and abstracted, but only those confirmed by qualified medical professionals were included in the analysis. For each psychiatric disorder, cumulative incidence rates for subjects with and without ADHD were estimated using the Kaplan-Meier method. Corresponding hazard ratios (HR) were estimated using Cox models adjusted for gender and mother's age and education at the subject's birth. The association between ADHD and the likelihood of having an internalizing or externalizing disorder was summarized by estimating odds ratios (OR). Attention-deficit/hyperactivity disorder was associated with a significantly increased risk of adjustment disorders (HR = 3.88), conduct/oppositional defiant disorder (HR = 9.54), mood disorders (HR = 3.67), anxiety disorders (HR = 2.94), tic disorders (HR = 6.53), eating disorders (HR = 5.68), personality disorders (HR = 5.80), and substance-related disorders (HR = 4.03). When psychiatric comorbidities were classified on the internalization-externalization dimension, ADHD was strongly associated with coexisting internalizing/externalizing (OR = 10.6), or externalizing-only (OR = 10.0) disorders.

This population-based study confirms that children with ADHD are at significantly increased risk for a wide range of psychiatric disorders. Besides treating the ADHD, clinicians should identify and provide appropriate treatment for psychiatric comorbidities.

Is the elimination of osteosarcoma cells with intraoperative "mesh autotransfusion" and leukocyte depletion filters possible?

Intraoperative autotransfusion is contraindicated in cancer surgery because of the possible risk of systemic tumor spread. The aim of the present study was to investigate whether a cell saver in combination with a white blood cell depletion filter can remove osteosarcoma cells. A defined number of osteosarcoma cells from an established cell line were added to red cell concentrates and Ringer solution. The tumor cell concentration was 1000/ ml in the first five experiments, 7111/ml in test no. 6, 1667/ml in test no. 7 and 167/ml in test no. 8. Following thorough mixing, each unit was processed separately by a cell saver (DIDECO BT 795/P) in its normal operation mode to produce a red cell concentrate. This red cell concentrate was filtered using a leukocyte depletion filter (PALL BPF 4). Samples were taken before and after processing with the autotransfuser and after filtration with the white cell depletion filter. Cytospin specimens from all samples were examined for osteosarcoma cells by three different methods (Papanicolaou stain, Vimentin antibodies, DNA analysis). After processing with the autotransfuser, tumor cells were identified in the red cell concentrate. No osteosarcoma cells were evident after the combined use of cell saver and leukocyte depletion filter.

The sole use of the autotransfuser DIDECO BT 795/P during osteosarcoma surgery is not recommended because of the potential danger of retransfusion of malignant cells. In combination with the leukocyte depletion filter PALL BPF 4, no osteosarcoma cells were identified in the red cell concentrate. Since the adhesiveness of tumor cells from established cell lines may be different from that of tumor cells in the intraoperative salvaged blood, further studies with blood from the surgical field are necessary to determine the efficacy of white cell depletion filters to eliminate osteosarcoma cells.

Doubling of serum creatinine: is it appropriate as the endpoint for CKD?

The evaluation of the progression of renal insufficiency, or decline in glomerular filtration rate (GFR), has been approached more simply and precisely by converting measured serum creatinine value into the reciprocal of serum creatinine, estimated GFR, or other parameters. Doubling of serum creatinine (simple doubling) is conveniently used as a surrogate endpoint for progression of renal disease but is thought to be biased unfairly by the initial value of serum creatinine (Scr(Int)). We proposed the definite decline in the reciprocal of serum creatinine (2-4 doubling) as a surrogate endpoint, comparing simple doubling with this new endpoint to verify the effect of Scr(Int) on the endpoint. For the purpose of comparison between endpoints, 194 patients in a historical cohort of chronic glomerulonephritis were investigated. Kaplan-Meier survival analysis was performed with the composite endpoint of need for dialysis and either simple doubling or 2-4 doubling. Then, the distribution of Scr(Int) was compared between total patients and patients developing each endpoint. The endpoint value of serum creatinine (Scr(End)) with 2-4 doubling was lower than that with simple doubling at Scr(Int)<2.00 mg/dl, and the difference of Scr(End) between simple doubling and 2-4 doubling was larger, as Scr(Int) became lower. In patients reaching simple doubling, Scr(Int) was higher than that of the total patients (1.66 vs. 1.07 mg/dl in median, respectively; p<0.001). In patients reaching 2-4 doubling, there was no significant difference in Scr(Int).

Patients with low serum creatinine concentration at baseline had a tendency of prolonged development into simple doubling. In contrast, with 2-4 doubling, there was no bias of Scr(Int).

Does short-term protein and energy supplementation activate nitrogen kinetics and accretion in poorly nourished elderly subjects?

An increase in protein intake exerts a stimulating effect on protein kinetics in children, young adults, and healthy elderly persons. However, there are few data on the response to such dietary changes in malnourished elderly subjects, despite important medical implications in this population. The objective of this study was to determine the metabolic response to short-term nutritional supplementation in moderately malnourished elderly subjects. The influence of 10 d of supplementation (1.67 MJ/d and 30 g protein/d) on body composition, resting energy expenditure, and whole-body protein kinetics was studied in 17 malnourished elderly patients and 12 healthy young adults. A control group of 6 malnourished elderly patients received no supplementation. Supplemented elderly subjects had a significantly greater fat-free mass gain than did unsupplemented elderly subjects (1.3 and 0.1 kg, respectively; age effect, P < 0.05; diet effect, P < 0.02) and a significantly greater increase in fasting rate of protein synthesis than did young supplemented subjects (0.6 and 0.2 g*kg FFM(-1)*11 h(-1); age effect, P < 0.05). The net protein balance in the supplemented elderly subjects in the fed state was positively correlated with protein intake (r(2) = 0.46) and in the fasted state was negatively correlated with protein intake (r(2) = 0.27). The sum of these regressions is a line with increasingly positive net diurnal protein balance produced by increasing protein intake.

These data provide evidence of a short-term anabolic response of protein metabolism to dietary supplementation in malnourished elderly patients that is likely to improve muscle strength and functional status.

Is oXA-181 Beta Lactamase a Major Mediator of Carbapenem Resistance in Enterobacteriaceae?

Detection of carbapenem hydrolyzing class D beta lactamase OXA-181, (a variant of OXA-48) in Enterobacteriaceae, is important, to institute appropriate therapy and to initiate preventive measures. This study was done to determine the presence of OXA 48 and its derivative OXA-181 in Enterobacteriaceae of pathogenic significance. One hundred and eleven non-repetitive Enterobacteriaceae isolates which were resistant to any of the cephalosporin subclasses III and which exhibited reduced susceptibility to carbapenems were included in the study. Minimum inhibitory concentrations (MICs) to imipenem and meropenem was determined by broth microdilution. Production of carbapenamase was screened by Modified Hodge test (MHT). Polymerase Chain Reaction (PCR) was done to detect the presence of bla OXA-181 and bla OXA-48 .Coexistence of other carbapenemase encoding genes, namely, NDM-1, VIM, IMP and KPC were also looked for, by PCR. Of all the isolates which were tested, only 2 (1.8%) revealed the presence of OXA-181 and OXA-48. These were Klebsiella pneumoniae and Citrobacter freundii. MICs of imipenem and meropenem for Klebsiella pneumoniae were 128mg/l and 64 mg/l and for Citrobacter freundii, they were 32mg/l and 16mg/l respectively. MHT was positive in both isolates.

Production of OXA-48 / OXA-181 is not a major mechanism of carbapenem resistance. PCR is the gold standard for its routine identification in clinical microbiology laboratory.

Is yttrium-90 radioembolization a safe and effective treatment for unresectable hepatocellular carcinoma : a single centre experience of 45 consecutive patients?

There is controversy regarding the role of yttrium-90 (90Y) radioembolization in the management of advanced, unresectable hepatocellular carcinoma (HCC). Forty-five consecutive patients underwent resin-based 90Y radioembolization for unresectable, HCC between 2006 and 2013 in Sydney, Australia. All patients were followed up with imaging studies at regular intervals until death. Radiologic response was evaluated with the Response Criteria in Solid Tumors (RECIST) criteria. Clinical toxicities were prospectively recorded. Survival was calculated by the Kaplan-Meier method and potential prognostic variables were identified on univariate and multivariate analysis. Follow-up in the complete cohort was 7.8 (range, 0.1-41.8) months. The median survival after 90Y radioembolization was 27.7 months with a 36-month survival of 26%. By RECIST criteria of the 40 patients followed-up beyond 2 months, a complete response (CR) to treatment was observed in 1 patients (3%), partial response (PR) in 18 (45%), stable disease (SD) in 11 (22%) and progressive disease (PD) in 10 (25%). On multivariate analysis only radiological response to treatment was independently associated with improved survival: CR/PR to treatment vs. SD vs. PD; p < 0.001. Thirteen patients (29%) developed clinical toxicity after treatment; all complications were minor (grade I/II) and resolved without active intervention.

Radioembolization with 90Y is a safe and effective treatment for unresectable HCC.

Does hospital-skilled nursing facility referral linkage reduce readmission rates among Medicare patients receiving major surgery?

In the health reform era, rehospitalization after discharge may result in financial penalties to hospitals. The effect of increased hospital-skilled nursing facility (SNF) linkage on readmission reduction after surgery has not been explored. To determine whether enhanced hospital-SNF linkage, as measured by the proportion of surgical patients referred from a hospital to a particular SNF, would result in reduced 30-day readmission rates for surgical patients, we used national Medicare data (2011-2012) and evaluated patients who underwent 1 of 5 operative procedures (coronary artery bypass grafting [CABG], hip fracture repair, total hip arthroplasty, colectomy, or lumbar spine surgery). Initial evaluation was performed using regression modeling. Patient choice in SNF referral was adjusted for using instrumental variable (IV) analysis with distance between an individuals' home and the SNF as the IV. A strong negative correlation (P < .001) was observed between the proportion of selected surgical discharges received by a SNF and the rate of hospital readmission. Increasing the proportion of surgical discharges decreased the likelihood of rehospitalization (regression coefficient, -0.04; 95% CI, -0.07 to -0.02). These findings were preserved in IV analysis. Increasing hospital-SNF linkage was found to reduce significantly the likelihood of readmission for patients receiving lumbar spine surgery, CABG, and hip fracture repair.

The benefits of increased hospital-SNF linkage seem to include meaningful reductions in hospital readmission after surgery. Overall, a 10% increase in the proportion of surgical referrals to a particular SNF is estimated to decrease readmissions by 4%. This may impact hospital-SNF networks participating in risk-based reimbursement models.

Does intraoperative endosonography enhance laparoscopy-assisted colon pull-through for high imperforate anus?

The authors used ultrasonographic endoprobes during laparoscopy-assisted colon pull-through (LACPT) for the repair of high imperforate anus to confirm the pull-through canal was surrounded symmetrically by pelvic floor muscles. Six patients with high imperforate anus were treated by LACPT (mean age at LACPT, 8.2 months). An endoscopic (12-MHz, 2.5-mm in diameter) and proctoscopic (7.5-MHz, 12-mm in diameter) probe were inserted into the proposed route of dissection intraoperatively to measure the thickness of the surrounding muscle tissue at at least 3 levels: the external anal sphincter, the levator ani muscle sling, and the intervening muscle complex. The average thickness of the external anal sphincter was 2.3 +/- 0.4 mm anteriorly, 2.4 +/- 0.4 mm on the left, 2.4 +/- 0.5 mm posteriorly, and 2.6 +/- 0.6 mm on the right. The average thickness of the muscle complex was 2.3 +/- 0.6 mm anteriorly, 2.2 +/- 0.5 mm on the left, 2.1 +/- 0.4 mm posteriorly, and 2.2 +/- 0.5 mm on the right. The average thickness of the left crus of the levator ani muscle was 1.8 +/- 0.3 mm, the right crus was 1.9 +/- 0.4 mm, and the rim located posterior to the rectum was 2.0 +/- 0.3 mm. No statistically significant difference was found between the measurements taken at each level.

Intraoperative endosonography during LACPT can greatly enhance the precision of positioning the pull-through canal.

Does vitamin D mediate inhibition of epithelial ovarian cancer by modulating cytokines?

Vitamin D deficiency is reported to be involved in pathogenesis of ovarian cancer. But the mechanism is yet to be explored. An imbalance between Th1 and Th2 activity play a crucial role in pathogenesis of many cancers. The purpose of the study is to find out the Th1/Th2 status by estimating TNF-α (Th1 marker) and IL-4 (Th2 marker) in ovarian cancer cases and controls and to correlate these with serum vitamin D levels. A case-control study with 50 ovarian cancer cases and 50 healthy controls was conducted. The cytokines TNF-α and IL-4 were estimated by ELISA. Serum vitamin D was measured by electro-chemiluminescence immunoassay method. Median TNF-α levels (12.2 vs 6.2 pg/ml; p value<0.001) were significantly higher in ovarian cancer patients and mean IL-4 levels (2.22 ± 0.51 vs 2.99 ± 0.68 pg/ml; p value<0.05) were significantly lower as compared to those of controls. Levels of TNF-α and IL-4 did not vary significantly with clinical staging, and histological grading. Vitamin D levels were negatively correlated with TNF-α and positively correlated with IL-4.

Low vitamin D levels promotes Th1 activity increasing TNF-α levels and inhibits Th2 activity decreasing IL-4 levels in ovarian cancer. These low levels of vitamin D may induce pro-inflammatory micro ambience which might contribute to pathogenesis of ovarian cancer.

Does microproteinuria predict Organ Failure in Patients Presenting with Acute Pancreatitis?

The disease course of acute pancreatitis (AP) ranges from mild and self-limiting to severe inflammation, associated with significant morbidity and mortality. At present, there are no universally accepted and reliable predictors for severity. Microproteinuria has been associated with the presence of systemic inflammatory response syndrome as well as trauma, although its association with AP is not well understood. The aim of this study was to investigate the value of microproteinuria to predict development of organ failure in AP. Consecutive AP patients were prospectively enrolled. Urine samples were collected upon admission, 12-24 h after admission, and 3 months post-discharge for calculation of urine α1-microglobulin-, albumin-, IgG-, and IgM/creatinine ratios. Data regarding AP etiology, severity, and development of organ failure were registered. Overall, 92 AP patients were included (14 % with organ failure; 6 % with severe AP). The α1-microglobulin-, albumin-, and IgG/creatinine ratios correlated with high-sensitivity C-reactive protein 48 h after admission (r = 0.47-0.61, p < 0.001 for all). They were also significantly higher in patients with versus without organ failure (p < 0.05 for all). The α1-microglobulin/creatinine ratio upon admission predicted organ failure [adjusted odds ratio 1.286, 95 % confidence interval (CI) 1.024-1.614] with similar accuracy (AUROC 0.81, 95 % CI 0.69-0.94) as the more complex APACHE II score (AUROC 0.86, 95 % CI 0.70-1.00).

The α1-microglobulin/creatinine ratio upon presentation with AP is related to inflammation and predicts development of organ failure. Further studies are warranted to evaluate its potential usefulness in predicting outcome for AP patients.

Do Media Use and Physical Activity Compete in Adolescents?

The displacement hypothesis predicts that physical activity and media use compete in adolescents; however, findings are inconsistent. A more differentiated approach at determining the co-occurrence of physical activity and media use behaviors within subjects may be warranted. The aim of this study was to determine the co-occurrence of physical activity and media use by identifying clusters of adolescents with specific behavior patterns including physical activity in various settings (school, sports club, leisure time) and different types of media use (watching TV, playing console games, using PC / Internet). Cross-sectional data of 2,083 adolescents (11-17 years) from all over Germany were collected between 2009 and 2012 in the Motorik-Modul Study. Physical activity and media use were self-reported. Cluster analyses (Ward's method and K-means analysis) were used to identify behavior patterns of boys and girls separately. Eight clusters were identified for boys and seven for girls. The clusters demonstrated that a high proportion of boys (33%) as well as girls (42%) show low engagement in both physical activity and media use, irrespective of setting or type of media. Other adolescents are engaged in both behaviors, but either physical activity (35% of boys, 27% of girls) or media use (31% of boys and girls) predominates. These adolescents belong to different clusters, whereat in most clusters either one specific setting of physical activity or a specific combination of different types of media predominates.

The results of this study support to some extent the hypothesis that media use and physical activity compete: Very high media use occurred with low physical activity behavior, but very high activity levels co-occurred with considerable amounts of time using any media. There was no evidence that type of used media was related to physical activity levels, neither setting of physical activity was related to amount of media use in any pattern.

Does cordyceps Militaris alleviate Severity of Murine Acute Lung Injury Through miRNAs-Mediated CXCR2 Inhibition?

Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are lethal diseases in humans, and the current treatments have limited therapeutic effects. Cordyceps militaris (CM) is a caterpillar-grown traditional medicinal mushroom, and has been used as a natural invigorant for longevity, endurance, and vitality in China. Recently, purified extracts from CM have been shown to have beneficial effects on various diseases including cancer. Nevertheless, a role of CM in ALI has not been examined previously. Here, we used a bleomycin-induced ALI model to study the effects of CM on the severity of ALI in mice. The levels of CXCR2, a receptor for Interleukin 8 (IL-8) in pulmonary microvascular endothelial cells, were examined in different experimental groups. The levels of microRNA (miR)-1321 and miR-3188 were also examined in lung samples and in CM. Adeno-associated viruses carrying miR-1321 and miR-3188 were injected into bleomycin-treated mice for evaluation their effects on the severity of ALI. CM treatment significantly alleviated the severity of bleomycin-induced ALI in mice. The increases in lung CXCR2 by bleomycin were significantly reduced by CM at protein level, but not at mRNA level. CM contained high levels of 2 miRNAs (miR-1321 and miR-3188) that target 3'-UTR of CXCR2 mRNA to inhibit its expression. Overexpression of miR-1321 and miR-3188 in mouse lung through AAV-mediated gene therapy mimicked the effects of CM.

CM may alleviate severity of murine ALI through miRNAs-mediated CXCR2 inhibition.

Does inhibition of vascular ectonucleotidase activities by the pro-drugs ticlopidine and clopidogrel favour platelet aggregation?

After conversion to their active forms by the liver, ticlopidine and clopidogrel exert antiplatelet effects through irreversible inhibition of the P2Y₁₂ receptor. Concentrations of nucleotides such as ADP, the physiological agonist at platelet P2Y₁ and P2Y₁₂ receptors, are regulated by vascular ectonucleotidases, mainly nucleoside triphosphate diphosphohydrolase (NTPDase)1 and ecto-5'-nucleotidase. Here we evaluate the effect of these pro-drugs on vascular ectonucleotidase activity and on the natural function of these enzymes in regulating platelet aggregation. Nucleotidase assays were performed by HPLC and by P(i) determination, using human umbilical vein endothelial cells (HUVEC) and protein extracts from transfected COS-7 cells as sources of enzymes. Platelet aggregation was assayed using human platelet-rich plasma. Each pro-drug inhibited endothelial ectonucleotidase activities and decreased their ability to block platelet aggregation in vitro. At their therapeutic concentrations, ticlopidine (60 µM) and clopidogrel (20 µM) inhibited ADP hydrolysis by HUVEC by about 80%, and AMP hydrolysis by one-third. Accordingly, these compounds showed a mixed-type inhibition of recombinant human NTPDase1 with an apparent K(i) (K(i,app) ) of 10 µM (clopidogrel) and 14 µM (ticlopidine). Recombinant rat ecto-5'-nucleotidase, but not its human orthologue, was inhibited by ticlopidine with a K(i,app) of 4.5 mM.

These pro-drugs facilitated platelet aggregation via the inhibition of vascular NTPDase1 in vitro. Further studies should be performed to assess whether this effect also occurs in vivo, especially at the beginning of treatment, before sufficient levels of active metabolites are produced by the liver.

Does early hyperbaric oxygen therapy improve outcome for radiation-induced hemorrhagic cystitis?

To assess the clinical factors that affect the efficacy of hyperbaric oxygen (HBO2) therapy in treating radiation-induced hemorrhagic cystitis. HBO2 therapy is an effective treatment for radiation-induced hemorrhagic cystitis, with reported response rates ranging from 76% to 100%. The data from patients with radiation-induced hemorrhagic cystitis treated at our institution between May 1988 and December 2001 were reviewed retrospectively. All patients received HBO2 therapy at 2.36 atm absolute pressure, with 90 minutes of 100% oxygen breathing per treatment. The outcome was assessed after at least 12 months of follow-up. We evaluated patient demographics, types of pelvic malignancy and radiotherapy, total radiation dose, onset and severity of hematuria, and prior intravesical management. Clinical improvement was defined as the absence of, or reduction in, macroscopic hematuria. A total of 60 patients (55 men and 5 women), mean age 70 years, received an average of 33 HBO2 treatments (range 9 to 63). Of the 60 patients, 48 (80%) had either total or partial resolution of hematuria. When treated within 6 months of hematuria onset, 96% (27 of 28) had complete or partial symptomatic resolution (P = 0.003). All 11 patients with previous clot retention had clinical improvement if treated within 6 months of hematuria onset (P = 0.007). Prior intravesical chemical instillation did not affect the clinical outcome. Patients who had undergone primary, adjuvant, or salvage external beam pelvic radiotherapy showed response rates of 81%, 83%, and 78%, respectively (P = 0.950).

Our results show that delivery of HBO2 therapy within 6 months of hematuria onset is associated with a greater therapeutic response rate. Treatment efficacy was independent of prior intravesical therapy and the timing of radiotherapy.

Is increased expression of pleiotrophin a prognostic marker for patients with gastric cancer?

BACKGROUND/AIMs: Pleiotrophin (PTN) have been demonstrated to play an important role in the development of human gastric cancer. However, the prognostic value remains unclear. The aim of this study was to investigate whether expression of PTN has prognostic relevance in human gastric cancer. Immunohistochemistry was used to investigate the expression of PTN proteins in 178 patients with gastric cancer. The level of PTN mRNA in gastric cancer tissues and paratumor tissues were evaluated in 52 paired cases by quantitative real-time polymerase chainreaction(qRT-PCR). Survival analysis by the Kaplan-Meier method was performed to assess prognostic significance. The expression level of PTN in gastric cancer tissues was significantly higher (P<0.001) than those in paratumor tissues according to the immunohistochemistry analysis, which was confirmed by qRT-PCR analysis. Additionally, the overexpression of PTN was significantly associated with the tumor site (P=0.001), Lauren’s classification (P<0.001),histologic differentiation(P=0.014),depth of invasion(P<0.001), TNM stage (P=0.003),and lymph node metastasis (P=0.002). Moreover, the Cox proportional- hazards regression analysis revealed that the increased expression of PTN was an independent prognostic factor for poor recurrence-free survival(RFS) and overall survival(OS)(both P<0.001).

These findings indicated that the expression of PTN is significantly correlated with prognosis in gastric cancer patients, suggesting that the expression of PTN may be used as an independent prognostic marker.

Post-sternotomy chronic osteomyelitis: is sternal resection always necessary?

The goal of this study was to investigate alternative strategies to the sternal resection in the treatment of post-sternotomy osteomyelitis. We report our experience in the treatment of chronic infection of median sternotomy following open heart surgery without sternal resection. A 4-year retrospective study was performed, consisting of 70 patients affected by post-sternotomy sternocutaneous fistulas due to chronic osteomyelitis: 45 patients underwent only medical treatment and 25 underwent steel wire removal and surgical debridement (conservative surgery). Of the 25, 7 patients underwent an additional vacuum assisted closure (VAC) therapy due to widespread infected subcutaneous tissue. The diagnosis of osteomyelitis was supported via 3D CT scan images. Complete wound healing was achieved in 67 patients including a patient who achieved healing after being affected by a fistula for over 24 years before coming under our observation, another, affected by mycobacteria other than tuberculosis osteomyelitis, who needed antimicrobial treatment for a period of 30 months and 2 who were affected by Aspergillus infection and needed radical cartilage removal. Fistula relapses were observed in 6 patients of the total 70, possibly due to the too short-term antibiotic therapy used in the presence of coagulase-negative Staphylococcus (CoNS) with multiple resistances and in the presence of Corynebacterium species.

Post-sternotomy chronic osteomyelitis can be successfully treated mainly by systemic antimicrobial therapy alone, without mandatory surgical treatments, provided that accurate microbiological and radiological studies are performed. The presence of CoNS and Corynebacterium species seemed to be associated with a need for a prolonged combined antimicrobial therapy with a minimum of 6 months up to a maximum of 18 months. The CT scan and the 3D reconstruction of the sternum proved to be a good method to evaluate the status of the sternum and support the treatments. The VAC therapy was not useful in treating osteomyelitis, although, if used appropriately in the postoperative deep sternal wound infection with the sponge fitted between the sternal edges, it seems to be an effective method to eradicate the infection in the sternum and to prevent chronic osteomyelitis.

Utilisation of coronary angiography after acute myocardial infarction in Ontario over time: have referral patterns changed?

To examine how physicians in Ontario, Canada, have altered their referral patterns for coronary angiography after acute myocardial infarction (AMI) over time. Retrospective analysis of multilinked administrative data. Province of Ontario, Canada. 146 365 Ontario AMI patients hospitalised between 1 April 1992 and 31 March 1999. Utilisation trends of coronary angiography among all patients, as well as within six subgroups: elderly (versus young), women (versus men), high (versus low) risk of 30 day mortality, high (versus low) socioeconomic status, cardiology (versus non-cardiology) attending physician specialty, and hospitals with (versus without) onsite revascularisation capacity. Cox proportional hazard models were adjusted for variations in patient, physician, and hospital characteristics over time. Angiography rates in Ontario increased from 23.2% in 1992 to 35.5% in 1999 (p<0.0001). Increases in utilisation of coronary angiography were most pronounced among the elderly (12.4-24.3% v 39.3-54.4% for non-elderly patients, p<0.0001), the affluent (24.6-38.7% v 22.0-32.3% for less affluent patients, p = 0.01), and those tended to by cardiologists (32.0-47.1% v 20.3-30.1% for non-cardiology attending specialties, p<0.0001) after adjusting for changes in baseline patient, physician, and hospital characteristics over time.

Despite universal health care availability, not all patients benefited equally from increases in service capacity for coronary angiography after AMI in Ontario. Wider implementation of data monitoring and explicit management systems may be required to ensure that appropriate utilisation of cardiac services is allocated to patients who are most in need.

Do quorum sensing regulation of the two hcp alleles in Vibrio cholerae O1 strains?

The type VI secretion system (T6SS) has emerged as a protein secretion system important to several gram-negative bacterial species. One of the common components of the system is Hcp, initially described as a hemolysin co-regulated protein in a serotype O17 strain of Vibrio cholerae. Homologs to V. cholerae hcp genes have been found in all characterized type VI secretion systems and they are present also in the serotype O1 strains of V. cholerae that are the cause of cholera diseases but seemed to have non-functional T6SS. The serotype O1 V. cholerae strain A1552 was shown to express detectable levels of Hcp as determined by immunoblot analyses using polyclonal anti-Hcp antiserum. We found that the expression of Hcp was growth phase dependent. The levels of Hcp in quorum sensing deficient mutants of V. cholerae were compared with the levels in wild type V. cholerae O1 strain A1552. The expression of Hcp was positively and negatively regulated by the quorum sensing regulators HapR and LuxO, respectively. In addition, we observed that expression of Hcp was dependent on the cAMP-CRP global transcriptional regulatory complex and required the RpoN sigma factor.

Our results show that serotype O1 strains of V. cholerae do express Hcp which is regarded as one of the important T6SS components and is one of the secreted substrates in non-O1 non-O139 V. cholerae isolates. We found that expression of Hcp was strictly regulated by the quorum sensing system in the V. cholerae O1 strain. In addition, the expression of Hcp required the alternative sigma factor RpoN and the cAMP-CRP global regulatory complex. Interestingly, the environmental isolates of V. cholerae O1 strains that showed higher levels of the HapR quorum sensing regulator in comparison with our laboratory standard serotype O1 strain A1552 where also expressing higher levels of Hcp.

Is maternal serum CA-125 level elevated in severe preeclampsia?

The aim of this study was to determine the relationship between serum concentrations of cancer antigen-125 (CA-125) and pre-eclampsia severity. We evaluated 91 females with a singleton pregnancy. Serum CA-125 levels were measured in subjects with severe pre-eclampsia (n=34) and those with mild pre-eclampsia (n=24). Females with healthy pregnancies (n=31) served as the control group. The three study groups were statistically similar in terms of maternal age, gestational age, and body mass index. The CA-125 level was significantly higher in the severe pre-eclampsia group than that in the mild pre-eclampsia and control groups (p<0.05). No significant difference in CA-125 levels between the mild pre-eclampsia and control groups was observed. CA-125 level was positively correlated with proteinuria (r=0.489, p=0.000), systolic blood pressure (r=0.503, p=0.018), and diastolic blood pressure (r=0.532, p=0.000). In contrast, CA-125 was negatively correlated with birth weight (r=0.266, p=0.012) and gestational age at birth (r=0.250, p=0.018).

CA-125 level increased in severe pre-eclampsia, which reflected abnormal trophoblastic invasion and chronic inflammation. Elevated levels of CA-125 in pre-eclamptic patients may be a marker of the disease severity.

p.Ala541Thr variant of MEN1 gene: a non deleterious polymorphism or a pathogenic mutation?

Multiple Endocrine Neoplasia Type 1 (MEN1) is an autosomal dominant inherited syndrome, related to mutations in the MEN1 gene. Controversial data suggest that the nonsynonymous p.Ala541Thr variant, usually considered as a non-pathogenic polymorphism, may be associated with an increased risk of MEN1-related lesions in carriers. The aim of this study was to evaluate the pathogenic influence of the p.Ala541Thr variant on clinical and functional outcomes. We analysed a series of 55 index patients carrying the p.Ala541Thr variant. Their clinical profile was compared to that of 117 MEN1 patients. The biological impact of the p.Ala541Thr variant on cell growth was additionally investigated on menin-deficient Leydig cell tumour (LCT)10 cells generated from Men1+/Men1- heterozygous knock-out mice, and compared with wild type (WT). The mean age at first appearance of endocrine lesions was similar in both p.Ala541Thr carriers and MEN1 patients, but no p.Ala541Thr patient had more than one cardinal MEN1 lesion at initial diagnosis. A second MEN1 lesion was diagnosed in 13% of MEN1 patients and in 7% of p.Ala541Thr carriers in the year following preliminary diagnosis. Functional studies on LCT10 cells showed that overexpression of the p.Ala541Thr variant did not inhibit cell growth, which is in direct contrast to results obtained from investigation of WT menin protein.

Taken together, these data raise the question of a potential pathogenicity of the p.Ala541Thr missense variant of menin that commonly occurs within the general population. Additional studies are required to investigate whether it may be involved in a low-penetrance MEN1 phenotype.

Is c-erbB-2 Oncoprotein expression associated with poor prognosis in squamous cell carcinoma of the cervix?

A polyclonal antihuman c-erbB-2 oncoprotein antibody recognized c-erbB-2 oncoprotein in routinely formaldehyde-fixed, paraffin-embedded specimens. Specimens taken from 192 patients with Stage III squamous cell carcinoma of the cervix treated with radiation therapy alone were investigated for c-erbB-2 oncoprotein expression using an immunohistochemical method. Cancer cells that were positive for c-erbB-2 oncoprotein showed a surface membrane staining pattern. Of the 192 patients, 143 were negative for c-erbB-2 oncoprotein, 12 were weakly positive or ambiguous, 31 were positive, and 6 were strongly positive. The 5-year survival rate of the 155 patients who tested c-erbB-2 negative or weakly positive was significantly better than that of the 37 patients whose results were positive or strongly positive (61% versus 41%, P = 0.022).

c-erbB-2 Oncoprotein expression in cancer cells may imply a poor prognosis for patients with Stage III squamous cell carcinoma of the cervix treated with radiation therapy alone.

Do pre-S mutations of hepatitis B virus affect genome replication and expression of surface antigens?

In chronic hepatitis B virus (HBV) infection, quantitative HBV surface antigen (qHBsAg) is useful for monitoring viral replication and treatment responses. We aimed to determine whether pre-S mutations have any effect on circulating qHBsAg. Plasmids expressing 1–8 amino acid deletion in pre-S1 ("pre-S1Δ1-8") and 3-25 amino acid deletion in pre-S2 ("pre-S2Δ3-25") were constructed. At 72 h posttransfection into Huh7 cells, qHBsAg were measured using electrochemiluminescence immunoassay analyzer. To mimic milieus of quasispecies, we co-transfected either pre-S1Δ1-8 or pre-S2Δ3-25 with wild type (WT). Pre-S mutations affected transcription and replication ability of HBV because of altered overlapping polymerase. Compared with WT, extracellular qHBsAg in pre-S1Δ1-8 and pre-S2Δ3-25 were on average 3.87-fold higher and 0.92-fold lower, respectively, whereas intracellular qHBsAg in pre-S1Δ1-8 and pre-S2Δ3-25 were 0.57-fold lower and 1.60-fold higher, respectively. Immunofluorescence staining of cellular HBsAg showed that pre-S1Δ1-8 had less staining and that pre-S2Δ3-25 had denser staining. As ratios of either pre-S1Δ1-8 or pre-S2Δ3-25:WT increased from 0:10 to 10:0 gradually, relative extracellular qHBsAg increased from 1.0 to 3.85 in pre-S1Δ1-8 co-transfection, whereas those decreased from 1.0 to 0.88 in pre-S2Δ3-25 co-transfection.

Pre-S mutations exhibit different phenotypes of genome replication and HBsAg expression according to their locations. Thus, qHBsAg level for diagnosis and prognostification in chronic HBV infection should be used more cautiously, considering emergences of pre-S deletion mutants.

Is new cardioprotective agent K201 natriuretic and glomerular filtration rate enhancing?

K201 (JTV519) is a newly developed 1,4-benzothiazepine drug with antiarrhythmic and cardioprotective properties. It functions via stabilization of the ryanodine receptor-calcium release channel in the heart (RyR2). This receptor has been identified in the kidney, and in vitro studies suggest a role in the control of renal hemodynamics. To date, the in vivo function of this receptor is undefined. We hypothesized that this new drug, which is being developed for the treatment of heart failure for its myocardial actions, also possesses renal hemodynamic enhancing and excretory properties. We also used immunohistochemistry to identify RyR2 in the normal canine kidney. We investigated the renal actions of K201 during intrarenal infusion in normal anesthetized dogs. K201 was infused after baseline measurements at 2 doses (0.1 and 0.5 mg.kg(-1).min(-1)). Immunohistochemistry was used to identify RyR2 presence in the kidney not exposed to K201. K201 was potently natriuretic and diuretic, with glomerular filtration rate- and renal blood flow-enhancing actions. The excretory responses to K201 administration were associated with decreases in distal tubular reabsorption of sodium despite a mild decrease in mean arterial pressure, which returned to baseline levels after K201 discontinuation. Immunohistochemistry of the normal canine kidney revealed the presence of RyR2 in the medullary collecting duct cells.

We report for the first time that the newly developed cardioprotective drug K201 possesses natriuretic, diuretic, glomerular filtration rate-enhancing, and vasodilating properties that go beyond myocardial actions and may support its therapeutic role in treatment of heart failure.

Does dickkopf-1 promote the differentiation and adipocytokines secretion via canonical Wnt signaling pathway in primary cultured human preadipocytes?

Dickkopf-1, a newly recognized antagonist for canonical Wnt signaling, is secreted in the early stage of human adipose-derived stem cells (ASCs) adipogenic differentiation. This study was aimed to investigate whether human recombinant DKK-1 (rhDKK-1) could affect the differentiation and metabolism as well as adipocytokines secretion in primary cultured human ASCs. Human ASCs were isolated from omental adipose tissue and induced to adipogenic differentiation in the absence or presence of Wnt signaling antagonist rhDKK-1 and agonist SB216763, respectively. mRNA and protein expression profiles of adipogenic factors during the differentiation process were analyzed using quantitative RT-PCR and Western blotting. Adipocytokines secretion levels in the culture medium were measured by ELISA method. Our results showed that DKK-1 was already expressed during the early stage of adipogenesis and reached the peak on the 9th day. Exogenous rhDKK-1 exposure accelerated the differentiation by up-regulating PPAR-γ and C/EBP-α, down-regulating Wnt3a, Wnt10b and β-catenin, without affecting non-canonical Wnt signaling marker (Wnt5a). In addition, rhDKK-1 treatment increased the secretion of leptin, RBP4, TNF-α and adiponectin during differentiation. rhDKK-1 treatment also significantly increased the intracellular accumulation of lipids and lipolysis. Thus, Wnt signal pathway agonist SB216763 down-regulated DKK-1 transcriptional and secretion levels during adipogenic process.

Our results suggest that rhDKK-1 could promote ASCs differentiation and increase adipocytokines secretion via canonical Wnt signaling pathway.

The association between overweight and illegal drug consumption in adolescents: is there an underlying influence of the sociocultural environment?

The aims of the study were to: a) Examine the distribution of gender-stratified body mass index (BMI), eating attitudes and use of addictive substances, under the hypothesis of a confluent prevalence of weight abnormalities, eating disorders and substance abuse. b) Demonstrate the extent to which family, peer-related and psychosocial factors are common elements in categories of compulsive behaviour.METHODOLOGY/ In the present cross-sectional study, data were collected through self reported questionnaires administered to a large sample of 33,185 15-19 years old adolescents (ESPAD®Italia), divided into weight categories based on the BMI percentile distribution. Multinomial analyses were adopted to address the influence of social, family, leisure time factors, Eating Attitude Test (EAT26) on the association between weight categories and drug use. Recent drugs use was more frequent in overweight and underweight adolescents (p<0.05), especially in females. An EAT26 score ≥20 was more common in overweight adolescents. Multinomial analysis abolished the relationship between overweight and the use of most drugs, implicating self-esteem, parents' educational level, and friendships as mediators of the association. Within the overweight category, adolescents reporting recent drug use, showed greater frequency of having drug-abusing friends (∼80%), and severe problems with parents and school (∼30%) compared to overweight adolescents without recent drug use.

The frequent association of overweight and substance use and the presence of common underlying social factors, highlights the need for an interdisciplinary approach involving individual-focused treatment models as well as public health, social and environmental changes to reduce food- and substances-related problems.

Does early access to a neurologist reduce the rate of missed diagnosis in young strokes?

We hypothesized that the presence of an in-house neurologist or a neurology or emergency medicine (EM) residency is associated with a lower rate of missed stroke diagnosis and a greater use of thrombolytic therapy. The outpatient Young Stroke registry from our academic medical center was reviewed. Patients 16 to 50 years of age who presented with ischemic stroke were included. Information on presentation, acute therapy, and missed diagnosis was obtained. The presence of an EM or neurology residency at the presenting hospital was recorded. We also assessed whether hospital teaching status in these fields affected missed diagnosis rates, the use of thrombolysis, or stroke intervention. Ninety-three patients were included. Thirteen patients were misdiagnosed. In hospitals with and without a neurology residency, the missed diagnosis rate was 6.3% versus 18.0%, respectively (P=.21). Two patients were misdiagnosed in hospitals with a neurology residency, but neither had neurology consultations in the emergency department. If these cases are removed from our analysis, the rate of missed diagnosis with and without a neurology residency is 0% versus 20.6%, respectively (P=.008). Acute stroke therapy was administered in 17.9% of patients seen with an EM residency, compared to 2.7% without an EM residency (P=.046). With and without a neurology residency, acute stroke therapy was administered in 25% versus 8.2% of cases, respectively (P=.055).

Young adults with ischemic stroke seen at hospitals with a neurology residency had a lower missed diagnosis rate. The presence of an EM resident or a neurology teaching program was associated with a greater use of acute stroke therapies. These results support initiatives to triage young adults with suspected acute stroke to hospitals with access to neurologic expertise in the emergency department.

Does compound FLZ inhibit lipopolysaccharide-induced inflammatory effects via down-regulation of the TAK-IKK and TAK-JNK/p38MAPK pathways in RAW264.7 macrophages?

The aim of this study was to investigate the effect of the squamosamide derivative FLZ (N-2-(4-hydroxy-phenyl)-ethyl-2-(2,5-dimethoxy-phenyl)-3-(3-methoxy-4-hydroxy-phenyl)-acrylamide) on lipopolysaccharide (LPS)-induced inflammatory mediator production and the underlying mechanism in RAW264.7 macrophages. RAW264.7 cells were preincubated with non-toxic concentrations of compound FLZ (1, 5, and 10 micromol/L) for 30 min and then stimulated with 10 microg/L LPS. The production of nitric oxide (NO), the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), and the activation of nuclear factor kappa-B (NF-kappaB) and mitogen-activated protein kinase (MAPK) pathways were examined. FLZ significantly inhibited the LPS-induced production of NO, as well as the expression of iNOS and COX-2 at both the RNA and the protein levels in RAW264.7 cells. The LPS-induced increase in the DNA binding activity of NF-kappaB and activator protein 1 (AP-1), the nuclear translocation of NF-kappaB p65, the degradation of the inhibitory kappaBalpha protein (IkappaBalpha) and the phosphorylation of IkappaBalpha, IkappaB kinase (IKK) alpha/beta, c-Jun NH(2)-terminal kinase (JNK) and p38 MAPKs were all suppressed by FLZ. However, the phosphorylation of extracellular signal-regulated kinase (ERK) was not affected. Further study revealed that FLZ inhibited the phosphorylation of transforming growth factor-beta (TGF-beta)-activated kinase 1 (TAK1), which is an upstream signaling molecule required for IKKalpha/beta, JNK and p38 activation.

FLZ inhibited the LPS-induced production of inflammatory mediators at least partly through the downregulation of the TAK-IKK and TAK-JNK/p38MAPK pathways.

Do posterior fossa and spinal gangliogliomas form two distinct clinicopathologic and molecular subgroups?

Gangliogliomas are low-grade glioneuronal tumors of the central nervous system and the commonest cause of chronic intractable epilepsy. Most gangliogliomas (>70%) arise in the temporal lobe, and infratentorial tumors account for less than 10%. Posterior fossa gangliogliomas can have the features of a classic supratentorial tumor or a pilocytic astrocytoma with focal gangliocytic differentiation, and this observation led to the hypothesis tested in this study - gangliogliomas of the posterior fossa and spinal cord consist of two morphologic types that can be distinguished by specific genetic alterations. Histological review of 27 pediatric gangliogliomas from the posterior fossa and spinal cord indicated that they could be readily placed into two groups: classic gangliogliomas (group I; n = 16) and tumors that appeared largely as a pilocytic astrocytoma, but with foci of gangliocytic differentiation (group II; n = 11). Detailed radiological review, which was blind to morphologic assignment, identified a triad of features, hemorrhage, midline location, and the presence of cysts or necrosis, that distinguished the two morphological groups with a sensitivity of 91% and specificity of 100%. Molecular genetic analysis revealed BRAF duplication and a KIAA1549-BRAF fusion gene in 82% of group II tumors, but in none of the group I tumors, and a BRAF:p.V600E mutation in 43% of group I tumors, but in none of the group II tumors.

Our study provides support for a classification that would divide infratentorial gangliogliomas into two categories, (classic) gangliogliomas and pilocytic astrocytomas with gangliocytic differentiation, which have distinct morphological, radiological, and molecular characteristics.

Does 4-hydroxy-2-nonenal mediate genotoxicity and bystander effects caused by Enterococcus faecalis-infected macrophages?

Enterococcus faecalis is a human intestinal commensal that produces extracellular superoxide and promotes chromosome instability via macrophage-induced bystander effects. We investigated the ability of 4-hydroxy-2-nonenal (4-HNE), a diffusible breakdown product of ω-6 polyunsaturated fatty acids, to mediate these effects. 4-HNE was purified from E faecalis-infected macrophages; its genotoxicity was assessed in human colon cancer (HCT116) and primary murine colon epithelial (YAMC) cell lines. 4-HNE induced G(2)-M cell cycle arrest, led to formation γH2AX foci, and disrupted the mitotic spindle in both cell lines. Binucleate tetraploid cells that formed after incubation with 4-HNE were associated with the activation of stathmin and microtubule catastrophe. Silencing glutathione S-transferase α4, a scavenger of 4-HNE, increased the susceptibility of epithelial cells to 4-HNE-induced genotoxicity. Interleukin-10 knockout mice colonized with superoxide-producing E faecalis developed inflammation and colorectal cancer, whereas colonization with a superoxide-deficient strain resulted in inflammation but not cancer. 4-HNE-protein adducts were found in the lamina propria and macrophages in areas of colorectal inflammation.

4-HNE can act as an autochthonous mitotic spindle poison in normal colonic epithelial and colon cancer cells. This finding links the macrophage-induced bystander effects to colorectal carcinogenesis.

Do young adults with uncomplicated dengue fever need hospitalisation?

Approximately 80 percent of all notified cases of dengue infections in Singapore were hospitalised from 2000 to 2005. We aimed to determine if hospitalised dengue patients had significant morbidity and mortality, and if admissions were in accordance with previously-published admission criteria. The medical records of the first 20 patients with laboratory-confirmed dengue from two consecutive months in three time periods were retrospectively reviewed. Demographical, clinical and laboratory data on admission, during hospitalisation and on discharge, were compared. There were 120 patients with a mean age of 35 years. Males comprised 77 percent and foreign workers 51 percent. Of the published admission criteria, 33 percent had vomiting, 22 percent diarrhoea, 13 percent abdominal pain, 18 percent bleeding and one patient had hypotension. 30 percent were above the minimum platelet threshold of 80,000/microL, but 50 percent had safe levels of platelets between 50,000 and 80,000/microL. Dengue haemorrhagic fever occurred in 4 percent with no death. After admission, platelet nadir was below 20,000/microL in only 9 percent and below 10,000/microL in only 2 percent of cases. Bleeding did not correlate with platelet count. Medical referral to the hospital was significantly associated with thrombocytopenia, while self-referral was significantly associated with vomiting.

Severe adverse outcome among young adults with uncomplicated dengue fever is rare. Instead of hospitalisation, daily outpatient monitoring with symptomatic treatment and medical leave may be a safe and feasible alternative.

Do infants with major congenital anomalies have an excess of macrosomia?

Infants that develop congenital anomalies may also have an excess prevalence of macrosomia (birth weight>or =4,000 g). This may indicate that abnormalities of glycemic control play a role in the etiology of birth defects. This study was undertaken to determine whether all infants with congenital anomalies have an excess of macrosomia and whether it is confined to specific types of anomalies. A case-control study was conducted, comparing the birth weights of 8,226 infants with congenital anomalies ascertained by the Texas Birth Defects Monitoring Division with those of 965,965 infants without birth defects. Odds ratios were calculated to determine the association between birth weight and congenital anomalies, for 45 specific defects, and for all these defects combined. For all 45 defects combined, a significant association occurred only in the highest birth weight category. Infants with congenital anomalies were more likely than infants without birth defects to have a birth weight>or =4,500 g (OR = 1.65; 95% CI = 1.39-1.96). Infants born with ventricular septal defects, atrial septal defects, ventricular hypertrophy, or anomalies of the great vessels were 1.5-2.5 times more likely to weigh>or =4,000 g than were infants without birth defects. Based on small numbers, a stronger excess of macrosomia was observed for infants with encephalocele, holoprosencephaly, anomalies of the corpus callosum, preaxial polydactyly, and omphalocele.

Our data suggest that infants with specific congenital anomalies are more likely to be macrosomic than are infants without an anomaly. If these findings are confirmed, associations between macrosomia and specific types of birth defects may help to identify birth defects that are caused by alterations in glycemic control.

Is podocyte foot process effacement in very early phase of passive Heymann nephritis a prerequisite for proteinuria?

Although foot process effacement is a characteristic alteration of podocytes in the proteinuric state, whether this is the cause or the result of proteinuria is not understood. We studied the morphology and molecular background of foot process effacement in relation to proteinuria, using the passive Heymann nephritis (PHN) model. Foot process effacement was evaluated by electron microscopy. C3 deposition and the expression of alpha 3-integrin, a major adhesion molecule of podocytes, and actin cytoskeleton were examined by immunofluorescent staining. alpha 3-Integrin was also evaluated by immunoelectron microscopy. Western blotting was performed to examine whether anti-Fx1A recognizes alpha 3 beta 1-integrin. Foot process effacement accompanied by decreased expression of alpha 3-integrin was already observed from day 1 after the injection of anti-Fx1A, but albuminuria was not observed until day 5. Complement activation, a key pathogenesis in PHN, was estimated to occur from day 2 after the appearance of foot process effacement. The degree of foot process effacement had not changed before the onset of albuminuria, while after the onset of albuminuria it significantly deteriorated with increased expression of actin. By immunoelectron microscopy, alpha 3-integrin decreased exclusively at the site of deposits. Western blotting showed anti-Fx1A recognizing beta1-integrin.

These findings indicate that complement-independent foot process effacement related to decreased expression of alpha 3 beta 1-integrin in a very early phase of PHN is not a prerequisite for proteinuria, and the deterioration of foot process effacement related to actin reorganization after the onset of albuminuria might be a secondary response to proteinuria.

Are desmoplakin and talin2 novel mRNA targets of fragile X-related protein-1 in cardiac muscle?

The proper function of cardiac muscle requires the precise assembly and interactions of numerous cytoskeletal and regulatory proteins into specialized structures that orchestrate contraction and force transmission. Evidence suggests that posttranscriptional regulation is critical for muscle function, but the mechanisms involved remain understudied. To investigate the molecular mechanisms and targets of the muscle-specific fragile X mental retardation, autosomal homolog 1 (FXR1), an RNA binding protein whose loss leads to perinatal lethality in mice and cardiomyopathy in zebrafish. Using RNA immunoprecipitation approaches we found that desmoplakin and talin2 mRNAs associate with FXR1 in a complex. In vitro assays indicate that FXR1 binds these mRNA targets directly and represses their translation. Fxr1 KO hearts exhibit an up-regulation of desmoplakin and talin2 proteins, which is accompanied by severe disruption of desmosome as well as costamere architecture and composition in the heart, as determined by electron microscopy and deconvolution immunofluorescence analysis.

Our findings reveal the first direct mRNA targets of FXR1 in striated muscle and support translational repression as a novel mechanism for regulating heart muscle development and function, in particular the assembly of specialized cytoskeletal structures.

Early rheumatoid arthritis: does gender influence disease expression?

To investigate whether gender is an independent factor associated with disease expression in early rheumatoid arthritis (RA) patients. 438 patients with early RA (disease duration less than one year) were studied. They all were patients with early RA who presented at the Rheumatology Clinic of the University Hospital of Ioannina during the period 1991-2000. All patients fulfilled the American College of Rheumatology criteria for RA. The demographic, clinical, laboratory, radiological and therapeutic characteristics of the disease at diagnosis, and at the last follow-up were analyzed according to gender. We studied 312 women and 126 men with early RA. The female to male ratio was 2.5:1 and the mean age at diagnosis was 49.4 +/- 14.9 years for women and 55.3 +/-15.6 years for men (P<0.0003). Women had a longer duration of follow-up (P<0.0003). There were no differences between genders in the general symptoms or the simmetricity of joint involvement at at disease onset. However at disease onset women had a higher erythrocyte sedimentation rate (ESR) (>30 mm/1st hour), although there were no significant differences between the two groups concerninig the rest of the clinical, laboratory and radiological findings. At the last follow-up women still had a higher ESR (>30 min/1st hour), but no significant differences were found between the two groups concerning the rest of the parameters investigated independently of the follow-up duration. Finally, women and men showed the same degree of radiological changes and functional ability and were treated similarly except for the more frequent use of hydroxychloroquine in women.

It seems that gender does not signficantly influence the expression of RA.

Answering questions at the point of care: do residents practice EBM or manage information sources?

To determine the types of information sources that evidence-based medicine (EBM)-trained, family medicine residents use to answer clinical questions at the point of care, to assess whether the sources are evidence-based, and to provide suggestions for more effective information-management strategies in residency training. In 2005, trained medical students directly observed (for two half-days per physician) how 25 third-year family medicine residents retrieved information to answer clinical questions arising at the point of care and documented the type and name of each source, the retrieval location, and the estimated time spent consulting the source. An end-of-study questionnaire asked 37 full-time faculty and the participating residents about the best information sources available, subscriptions owned, why they use a personal digital assistant (PDA) to practice medicine, and their experience in preventing medical errors using a PDA. Forty-four percent of questions were answered by attending physicians, 23% by consulting PDAs, and 20% from books. Seventy-two percent of questions were answered within two minutes. Residents rated UptoDate as the best source for evidence-based information, but they used this source only five times. PDAs were used because of ease of use, time factors, and accessibility. All examples of medical errors discovered or prevented with PDA programs were medication related. None of the participants' residencies required the use of a specific medical information resource.

The results support the Agency for Health Care Research and Quality's call for medical system improvements at the point of care. Additionally, it may be necessary to teach residents better information-management skills in addition to EBM skills.

Does cytotoxicity of prostaglandin analog eye drop preserved with benzalkonium chloride in multiple corneoconjunctival cell lines?

This study evaluated the cytotoxicity of five prostaglandin analog ophthalmic solutions on four ocular surface cell lines, ie, Chang (human conjunctiva), SIRC (rabbit cornea), RC-1 (rabbit cornea), and BCE C/D-1b (bovine cornea). Cell viability was measured by neutral red and MTT assays in cells treated for 10, 30, or 60 minutes with various doses of prostaglandins (undiluted, and 2- and 10-fold dilutions). The number of cell lines with viability >/=50% in the presence of selected dilution of the drug (CVS(50)) was used for comparison. In addition, 24 cell viability comparisons (four cell lines, two assays, and three exposure times) were made between latanoprost (Xalatan((R))) and each other solution at each dose. A comparison between the newly introduced tafluprost (Tapros((R))) with 0.01% benzalkonium chloride was also made. The order of cell viability determined by CVS(50) was Travatan Z((R)) (travoprost with the SofZia system) > Tapros >/= Travatan((R)) (travoprost) = Xalatan > Rescula((R)) (unoproston). This was consistent with the results of direct comparisons between Xalatan and the other drugs. There was no clear difference in cell viability between Tapros and benzalkonium chloride.

Use of two assays, multiple cell lines, and various dilutions and exposure times provided a unique evaluation of cytotoxicity among ophthalmic solutions. CVS(50) was useful for comparison of the cell viability of the solutions.

Is generalized Joint Laxity Associated with Primary Occurrence and Treatment Outcome of Lumbar Disc Herniation?

We investigated relationships between generalized joint laxity and primary lumbar disc herniation occurrence and compared clinical outcomes after conservative treatment in lumbar disc herniation patients with and without generalized joint laxity. The study group included 128 men, and the control group included 276 men matched for age and body mass index with the study group. The primary outcome measure was the presence or absence of generalized joint laxity using the Beighton scale. Clinical outcomes measured by the visual analog scale and the Oswestry disability index 2 years after conservative treatment were the secondary outcome measure. Generalized joint laxity prevalence was 13.2% in the study group and 5.1% in the control group, a significant difference (P=0.01). Spearman correlation analysis revealed that weight (r=0.162, P=0.03), body mass index (r=0.131, P=0.03), and generalized joint laxity (r=0.372, P<0.01) significantly correlated with lumbar disc herniation occurrence. In multivariate regression analysis, generalized joint laxity was the only significant lumbar disc herniation predictor (P=0.002; 95% confidence interval, 1.08 to 5.26). Generalized joint laxity in lumbar disc herniation patients was associated with worse clinical outcomes after conservative treatment measured by visual analog scale scores for lower extremity pain (P=0.02), lower back pain (P=0.03), and Oswestry disability index scores (P=0.03).

Generalized joint laxity might be associated with lumbar disc herniation occurrence and might also be a negative predictor of worse clinical outcomes after conservative treatment.

Does massage impair postexercise muscle blood flow and `` lactic acid '' removal?

This study tested the hypothesis that one of the ways sports massage aids muscle recovery from exercise is by increasing muscle blood flow to improve "lactic acid" removal. Twelve subjects performed 2 min of strenuous isometric handgrip (IHG) exercise at 40% maximum voluntary contraction to elevate forearm muscle lactic acid. Forearm blood flow (FBF; Doppler and Echo ultrasound of the brachial artery) and deep venous forearm blood lactate and H+ concentration ([La-], [H+]) were measured every minute for 10 min post-IHG under three conditions: passive (passive rest), active (rhythmic exercise at 10% maximum voluntary contraction), and massage (effleurage and pétrissage). Arterialized [La-] and [H+] from a superficial heated hand vein was measured at baseline. Data are presented as mean +/- SE. Venoarterial [La-] difference ([La-]v-a) at 30 s of post-IHG was the same across conditions (passive = 6.1 +/- 0.6 mmol x L(-1), active = 5.7 +/- 0.6 mmol x L(-1), massage = 5.5 +/- 0.6 mmol x L(-1), NS), whereas FBF was greater in passive (766 +/- 101 mL x min(-1)) versus active (614 +/- 62 mL x min(-1), P = 0.003) versus massage (540 +/- 60 mL x min(-1), P < 0.0001). Total FBF area under the curve (AUC) for 10 min after handgrip was significantly higher in passive versus massage (4203 +/- 531 vs 3178 +/- 304 mL, P = 0.024) but not versus active (3584 +/- 284 mL, P = 0.217). La(-)- efflux (FBF x [La-]v-a) AUC mirrored FBF AUC (passive = 20.5 +/- 2.8 mmol vs massage = 14.7 +/- 1.6 mmol, P = 0.03, vs active = 15.4 +/- 1.9 mmol, P = 0.064). H+ efflux (FBF x [H+]v-a) was greater in passive versus massage at 30 s (2.2 +/- 0.4e(-5) vs 1.3 +/- 0.2e(-5) mmol, P < 0.001) and 1.5 min (1.0 +/- 0.2e(-5) vs 0.6 +/- 0.09e(-5) mmol, P = 0.003) after IHG.

Massage impairs La(-) and H+ removal from muscle after strenuous exercise by mechanically impeding blood flow.

Do polymerization capacity of orthodontic composites analyzed by Fourier transform infrared spectroscopy?

The aim of this in-vitro study was to analyze the polymerization capacity of 5 orthodontic composites by determining the degree of monomer conversion (DC). Fourier transform infrared spectroscopy was used to evaluate the DC of the orthodontic composites immediately after polymerization and after storage in artificial saliva at 37°C ± 1°C for 30 days. The resin-based adhesive composites investigated were Bisco Ortho (Bisco, Schaumburg, Ill), Heliosit Orthodontics (Ivoclar, Schaan, Liechtenstein), Kurasper F (Kuraray, Okayama, Japan), Light Bond (Reliance Orthodontic Products, Itasca, Ill), and Transbond XT (3M Unitek, Monrovia, Calif), cured with Elipar FreeLight 2 (3M ESPE, St Paul, Minn) for the testing of the DC values. Fifty cylindrical specimens were manufactured in molds. The data were analyzed by 2-factor analysis of variance (ANOVA) and Tukey HSD test. According to 2-way ANOVA, the DC was significantly influenced by composite type (P <0.05); after 30 days, there were no differences among the composite types for the DC. The interaction of orthodontic composites and time played a statistically significant role in the DC (P <0.05), but there was no statistically significant influence of time for the DC (P >0.05).

The DC was found to change according to composite materials used, and Bisco Ortho showed the most DC performance. The DC of orthodontic composites is a complex process that is affected not only by inorganic filler content of the composite but also the monomer type and many other factors. Sufficient DC values of 5 commercially available orthodontic composites can be achieved with a new-generation light-emitting diode curing light.

Do intrinsically disordered domains deviate significantly from random sequences in mammalian proteins?

In order to characterize mammalian intrinsically disordered domains (IDDs) we examined the patterns in their amino acid abundance as well as overrepresented local sequence motifs. We considered IDDs from mouse proteins associated with innate immune responses as well as a set of generic human genes. These sets were compared with artificially generated random sequences with the same overall amino acid abundance and length distributions. IDDs were then clustered by amino acid abundance, and further analyzed in terms of co-occurrence of clusters with functionally characterized Pfam domains. Overall, IDDs were very different from randomly generated sequences. The deviation from random distributions was at least as great as that for ordered domains, for which the deviation can be rationalized in terms of strong evolutionary pressure for structure and function. The co-occurrence of certain Pfam domains with specific IDD clusters was found to be significant (p-value < 0.01). Local sequence motifs that were over-represented in the innate immune set consisted mostly of low complexity fragments, primarily characterized by amino acid repeats, and could not be assigned an obvious functional role.

Our results suggest that IDDs are constrained within a narrow subset of possible sequences. This is most likely a result of biophysical restraints that have yet to be elucidated. More detailed examination of the functional relationship between the IDDs and associated Pfam domains is one possible avenue of investigation.

Does systemic treatment of xenografts with vaccinia virus GLV-1h68 reveal the immunologic facet of oncolytic therapy?

GLV-1h68 is an attenuated recombinant vaccinia virus (VACV) that selectively colonizes established human xenografts inducing their complete regression. Here, we explored xenograft/VACV/host interactions in vivo adopting organism-specific expression arrays and tumor cell/VACV in vitro comparing VACV replication patterns. There were no clear-cut differences in vitro among responding and non-responding tumors, however, tumor rejection was associated in vivo with activation of interferon-stimulated genes (ISGs) and innate immune host's effector functions (IEFs) correlating with VACV colonization of the xenografts. These signatures precisely reproduce those observed in humans during immune-mediated tissue-specific destruction (TSD) that causes tumor or allograft rejection, autoimmunity or clearance of pathogens. We recently defined these common pathways in the "immunologic constant of rejection" hypothesis (ICR).

This study provides the first prospective validation of a universal mechanism associated with TSD. Thus, xenograft infection by oncolytic VACV, beyond offering a promising therapy of established cancers, may represent a reliable pre-clinical model to test therapeutic strategies aimed at modulating the central pathways leading to TSD; this information may lead to the identification of principles that could refine the treatment of cancer and chronic infection by immune stimulation or autoimmunity and allograft rejection through immune tolerance.

Does study of luminance effects on pinhole test result for visually impaired patients?

The visual acuity of visually impaired patients has been reported to improve after a refraction, despite pinhole test results that show a decline or no change in acuity. Our aim was to investigate whether the pinhole-induced reduction in retinal illuminance accounted for these unreliable predictions of best-corrected acuity. Participants were 64 adult patients referred for low-vision rehabilitation. Neutral density filters reproduced the pinhole-induced luminance loss, allowing pinhole test and postrefraction acuities to be measured at essentially equivalent levels of retinal illuminance. The following data were collected in random order from each subject's better eye: (1) habitual visual acuity, (2) habitual visual acuity with filter, (3) habitual visual acuity with pinhole, (4) best-corrected/postrefraction visual acuity, (5) postrefraction visual acuity with filter. On average, the pinhole test under-estimated postrefraction visual acuity by six letters (95% confidence limits = +/- 20). The pinhole test underestimated postrefraction visual acuity with the filter by two letters (95% confidence limits = +/- 16). Among subjects whose acuity improved with the pinhole test (N = 24), 83% experienced better postrefraction visual acuity. Among subjects whose acuity declined or remained unchanged with the pinhole test (N = 40), 50% achieved better postrefraction visual acuity.

The pinhole-induced luminance loss contributed to inadequate predictions of postrefraction visual acuity. Pinhole test results were enormously variable, underestimating and overestimating postrefraction visual acuity. The pinhole test was less reliable when improvements in postrefraction visual acuity were small. Visually impaired patients deserve periodic refractions, and the pinhole test result should not be used as a dichotomizer for clinical decisions regarding the need for a refraction.

Is right ventricular dysfunction in the R6/2 transgenic mouse model of Huntington 's disease unmasked by dobutamine?

Increasingly, evidence from studies in both animal models and patients suggests that cardiovascular dysfunction is important in HD. Previous studies measuring function of the left ventricle (LV) in the R6/2 model have found a clear cardiac abnormality, albeit with preserved LV systolic function. It was hypothesized that an impairment of RV function might play a role in this condition via mechanisms of ventricular interdependence. To investigate RV function in the R6/2 mouse model of Huntington's disease (HD). Cardiac cine-magnetic resonance imaging (MRI) was used to determine functional parameters in R6/2 mice. In a first experiment, these parameters were derived longitudinally to determine deterioration of cardiac function with disease progression. A second experiment compared the response to a stress test (using dobutamine) of wildtype and early-symptomatic R6/2 mice. There was progressive deterioration of RV systolic function with age in R6/2 mice. Furthermore, beta-adrenergic stimulation with dobutamine revealed RV dysfunction in R6/2 mice before any overt symptoms of the disease were apparent.

This work adds to accumulating evidence of cardiovascular dysfunction in R6/2 mice, describing for the first time the involvement of the right ventricle. Cardiovascular dysfunction should be considered, both when treatment strategies are being designed, and when searching for biomarkers for HD.

Does bacillus subtilis GlcK activity require cysteines within a motif that discriminates microbial glucokinases into two lineages?

Bacillus subtilis glucokinase (GlcK) (GenBank NP_390365) is an ATP-dependent kinase that phosphorylates glucose to glucose 6-phosphate. The GlcK protein has very low sequence identity (13.7%) to the Escherichia coli glucokinase (Glk) (GenBank P46880) and some other glucokinases (EC 2.7.1.2), yet glucose is merely its substrate. Our lab has previously isolated and characterized the glcK gene. Microbial glucokinases can be grouped into two different lineages. One of the lineages contains three conserved cysteine (C) residues in a CXCGX(2)GCXE motif. This motif is also present in the B. subtilis GlcK. The GlcK protein occurs in both monomer and homodimer. Each GlcK monomer has six cysteines. All cysteine residues have been mutated, one-by-one, into alanine (A). The in vivo GlcK enzymatic activity was assayed by functional complementation in E. coli UE26 (ptsG ptsM glk). Mutation of the three motif-specific residues led to an inactive enzyme. The other mutated forms retained, or in one case (GlcKC321A) even gained, activity. The fluorescence spectra of the GlcKC321A showed a red shift and enhanced fluorescence intensity compare to the wild type's.

Our results emphasize the necessity of cysteines within the CXCGX(2)GCXE motif for GlcK activity. On the other hand, the C321A mutation led to higher GlcKC321A enzymatic activity with respect to the wild type's, suggesting more adequate glucose phosphorylation.

Are meniscal tears and articular cartilage injury predictive of inferior patient outcome after surgical reconstruction for the dislocated knee?

A paucity of data exists on the effects of articular cartilage and meniscal injury in the setting of knee dislocations. The purpose of this study is to determine whether concomitant intra-articular injuries at the time of multiligament reconstruction for knee dislocation are associated with inferior outcomes. The records of patients who underwent surgical treatment for multiligament knee injury between 1992 and 2012 were retrospectively reviewed. Patients included had a PCL-based multiligament knee injury or a minimum of three disrupted ligaments, both indicative of knee dislocation. A logistic regression model was used to determine whether articular cartilage injuries (grade 2 involving ≥50 % of the condylar width or greater, or any grade III/IV lesions) and meniscus tears are predictors of IKDC outcome scores collected at a minimum of 2 years postoperatively. Of the 121 patients who met inclusion criteria, 2-year minimum follow-up was available on 95 patients (79 %). The cohort was 77 % male and had a median age of 32 years (16-62) at the time of surgery and was followed for an average of 6 years. Articular cartilage injury was present in 40 % of knees: medial femoral condyle (20 %); medial tibial plateau (9 %); lateral femoral condyle (5 %); lateral tibial plateau (4 %); patella (18 %); trochlear (5 %). Meniscal injury was present in 56 % of patients (isolated medial, 22 %; isolated lateral, 22 %; combined, 12 %). IKDC scores were significantly lower for patients with any cartilage damage (p = 0.03), combined medial and lateral meniscus tears (p = 0.02), medial-sided articular cartilage damage (p = 0.03), medial femoral condyle (p = 0.04) and trochlear (p = 0.03) lesions.

Articular cartilage damage and meniscus tears are frequently associated with a knee dislocation. This study showed IKDC scores were significantly lower for patients with cartilage damage or combined medial and lateral meniscus tears at mid-term follow-up of 6 years.

Liposuction of the arm concurrent with brachioplasty in the massive weight loss patient: is it safe?

Brachioplasty continues to be a sought-after procedure among the massive weight loss population. Residual adiposity of the upper arm can make this procedure more difficult. The authors sought to determine the safety of arm liposuction outside the region of excision with concomitant excisional brachioplasty. Data were analyzed from a prospective registry of massive weight loss patients who underwent brachioplasty alone or with concurrent arm liposuction. Variables examined included age, sex, body mass index, method of weight loss, medical comorbidities, and smoking status. Outcomes included complications such as seroma, wound dehiscence, infection, hematoma, lymphedema, and need for revision. Multivariate analyses were performed to assess outcome measures. One hundred forty-four patients (139 women and five men; mean body mass index, 29.6 ± 4.1 kg/m; mean age, 46 ± 10.7 years) underwent brachioplasty. Sixty-four patients had concomitant arm liposuction at the time of brachioplasty. The remaining 80 patients underwent excisional brachioplasty alone. Despite significantly higher operative body mass indices among those undergoing concurrent liposuction, no significant differences in complication rates were seen between the liposuction and excision-alone cohorts for seroma (19.1 percent versus 23.1 percent), wound dehiscence (7.9 percent versus 2.6 percent), infection (4.8 percent versus 6.4 percent), hematoma (3.2 percent versus 0 percent), or lymphedema (3.2 percent versus 1.3 percent). Revision rates were similar between the two groups (9.5 percent with liposuction and 8.9 percent without liposuction).

Liposuction can be performed safely and effectively outside the region of excision at the time of brachioplasty without the need for prior debulking or staged arm-contouring procedures.CLINICAL QUESTION/

Is nocturnal blood pressure non-dipping prevalent in severely obese , prepubertal and early pubertal children?

To investigate the prevalence of nocturnal blood pressure dipping among obese prepubertal and early pubertal children and to analyse the relationship between dipping and measures of insulin-glucose metabolism or sleep-disordered breathing. We studied 76 obese children (41% girls) under clinical care, with an average age of 10.4 ± 1.7 and a body mass index Z-score (BMI Z-score) of 6.2 ± 1.6. We performed a 24-h ambulatory blood pressure measurement. Non-dipping was defined as a nocturnal blood pressure reduction of <10%. We calculated measures of insulin-glucose metabolism from the performed frequently sampled intravenous glucose-tolerance test and from fasting blood samples. Overnight sleep polygraph recordings were performed to assess sleep-disordered breathing. Forty-two percent of the children were systolic non-dippers, and 17% were diastolic non-dippers. There were no associations between systolic or diastolic dipping and measures of insulin-glucose metabolism after adjustments for BMI Z-score, gender and pubertal status. There were no associations between dipping and measures of sleep-disordered breathing.

Nocturnal non-dipping was two times higher among severely obese, prepubertal and early pubertal children, compared to previous reports among children in general. There were no associations between nocturnal dipping and insulin-glucose metabolism or measures of sleep-disordered breathing in this group.

Are quadriphasic waveforms superior to triphasic waveforms for transthoracic defibrillation in a cardiac arrest swine model with high impedance?

We have demonstrated previously that triphasic waveform shocks were superior to biphasic waveform shocks for transthoracic defibrillation. Our purpose was to compare the efficacy and safety of quadriphasic versus triphasic shocks for transthoracic defibrillation in a porcine model. Sixteen adult swine (19-25 kg, mean: 21.5 kg) were deeply anesthetized and intubated. To simulate impedance of the human chest, fixed electrical resistors (25 or 50 ohms) was placed in series with the defibrillator and the chest of each pig. After 30 s of electrically induced VF, each pig received transthoracic shocks, using either a truncated exponential triphasic waveform (5 ms positive pulse duration, 5 ms negative pulse duration and 5 ms positive pulse duration, total waveform duration 15 ms) or a quadriphasic waveform (5/5/5/5 ms, total waveform duration 20 ms). Each pig received transthoracic triphasic and quadriphasic shocks at three selected energy levels (50, 100 and 150 J) in random sequence. Four shocks were delivered at each energy level to construct an energy versus % success curve. Success was defined as VF termination at 5 s after shock. The total shocks were divided into three groups based on the delivered energy actually delivered to the animal: <40, 40-65 and >65 J. Delivered energy = (animal impedance/total impedance) times selected energy of the shock. For high-impedance animals (86-102 ohms), quadriphasic waveform shocks achieved significantly higher percent shock success than triphasic shocks for the termination of VF at the energy levels of >65 J actually delivered (quadriphasic 72.7+/-12.2%, triphasic 38.9+/-7.7%, p<0.02). No differences in the shock success between quadriphasic and triphasic waveforms were found for other two energy levels. There were no differences in ventricular tachycardia or asystole after shocks between quadriphasic and triphasic waveforms.

In this porcine model, 20 ms (5/5/5/5) quadriphasic shocks were superior to 15 ms (5/5/5) triphasic shocks for transthoracic defibrillation in animals with impedances that simulated high impedance in humans.

Does transcriptome sequencing of lentil based on second-generation technology permit large-scale unigene assembly and SSR marker discovery?

Lentil (Lens culinaris Medik.) is a cool-season grain legume which provides a rich source of protein for human consumption. In terms of genomic resources, lentil is relatively underdeveloped, in comparison to other Fabaceae species, with limited available data. There is hence a significant need to enhance such resources in order to identify novel genes and alleles for molecular breeding to increase crop productivity and quality. Tissue-specific cDNA samples from six distinct lentil genotypes were sequenced using Roche 454 GS-FLX Titanium technology, generating c. 1.38 × 106 expressed sequence tags (ESTs). De novo assembly generated a total of 15,354 contigs and 68,715 singletons. The complete unigene set was sequence-analysed against genome drafts of the model legume species Medicago truncatula and Arabidopsis thaliana to identify 12,639, and 7,476 unique matches, respectively. When compared to the genome of Glycine max, a total of 20,419 unique hits were observed corresponding to c. 31% of the known gene space. A total of 25,592 lentil unigenes were subsequently annoated from GenBank. Simple sequence repeat (SSR)-containing ESTs were identified from consensus sequences and a total of 2,393 primer pairs were designed. A subset of 192 EST-SSR markers was screened for validation across a panel 12 cultivated lentil genotypes and one wild relative species. A total of 166 primer pairs obtained successful amplification, of which 47.5% detected genetic polymorphism.

A substantial collection of ESTs has been developed from sequence analysis of lentil genotypes using second-generation technology, permitting unigene definition across a broad range of functional categories. As well as providing resources for functional genomics studies, the unigene set has permitted significant enhancement of the number of publicly-available molecular genetic markers as tools for improvement of this species.

Does hormone administration promote the epithelium healing in patients with recurrent corneal epithelial exfoliation?

To investigate the effects of hormone administration upon epithelium healing in patients with recurrent corneal epithelial exfoliation. The recurrence rate of 56 patients with recurrent corneal epithelial exfoliation was compared after 3-month follow up, 30 patients of whom received the basic treatment of bFGF and pressure bandage plus prednisone administration (combination treatment group, ie. A) and the other 26 patients received the basic treatment alone (single treatment group, ie. B). No patients showed recurrence in the combination treatment group. But there were 20 patients (76.92%) in the basic treatment group recurred. χ² test showed that χ²=35.9. The two groups had significant difference regarding the recurrence of corneal epithelial exfoliation (P<0.01).

For the patients with recurrent corneal epithelial exfoliation, hormone administration should be considered to reduce the recurrence and protect the function of cornea as a supplement to the basic treatment.

Are pre-procedural Elevated White Blood Cell Count and Neutrophil-Lymphocyte ( N/L ) Ratio Predictors of Ventricular Arrhythmias During Percutaneous Coronary Intervention?

The absolute white blood cell (WBC) count and neutrophil to lymphocyte (N/L) ratio are predictors of death/myocardial infarction in patients who have undergone coronary angiography. We hypothesized that a pre-procedural elevated WBC count and an elevated N/L ratio would be a predictor of development of significant ventricular arrhythmias in subjects undergoing percutaneous coronary intervention (PCI). We retrieved the data for all patients developing ventricular arrhythmia during PCI between 1999 to 2009 from our cath lab database (from 30,798 records), a total of 70 patients (Group I), and tabulated their WBC counts and absolute neutrophil and lymphocyte counts as well as N/L ratios. We compared the data with a random group of age, gender, medications and pre-existing condition matched controls (n=70) (Group II). We also adjusted for amount of myocardium under jeopardy. Group I had a significantly higher total WBC count (means 14,344 Vs 6852; 95% CI; p=0.0004); neutrophil count (means 75.79% Vs 58.06%; 95% CI; p < 0.0001) and N/L ratio (means 3.79 Vs 1.56; 95% CI; p < 0.0001) [means compared with t test].

Our data suggests a pre-procedural elevated WBC count, neutrophils and elevated N/L ratio are predictors of significant ventricular arrhythmias in patients undergoing percutaneous coronary intervention (PCI).

Does a role for PHANTASTICA in medio-lateral regulation of adaxial domain development in tomato and tobacco leave?

Diverse leaf forms in nature can be categorized into two groups: simple and compound. A simple leaf has a single blade unit, whilst a compound leaf is dissected into leaflets. For both simple and compound leaves, a MYB domain transcription factor PHANTASTICA (PHAN) plays an important role in establishing the adaxial domain in the leaf. Absence of PHAN in arabidopsis and antirrhinum leaves supresses blade development, and in tomato suppresses leaflet development. However, in the rachis and petiole regions of tomato leaves where PHAN and the adaxial domain coexist, it has been unclear why leaf blade and leaflets are not formed. We hypothesized that PHAN regulates the medio-lateral extent of the adaxial domain, thereby determining compound leaf architecture. To test this hypothesis, we generated and analysed transgenic tomato plants expressing tomato PHAN (SlPHAN) under the Cauliflower mosaic virus (CaMV) 35S promoter in both sense and antisense orientations, and tobacco plants that over-express tomato SlPHAN. Modulations in SlPHAN resulted in a variety of leaf morphologies such as simple, ternate and compound in either a peltate or non-peltate arrangement. Measurements of the extent of the adaxial domain along the wild-type tomato leaf axis showed that the adaxial domain is narrowed in the rachis and petiole in comparison with regions where laminar tissue arises. In antiSlPHAN transgenic leaves, no blade or leaflet was formed where the adaxial domain was medio-laterally narrowed, and KNOX gene expression was correlatively upregulated. CaMV35S::SlPHAN expression led to widening of the adaxial domain and ectopic blade outgrowth in the rachis of tomato and in the petiole of tobacco. Taken together, these results suggest that SlPHAN plays a role in medio-lateral extension of the adaxial domain and contributes to final leaf morphology in tomato.

This study provides a novel insight into leaf architecture in tomato and highlights how changes in the expression domain of a master regulator gene such as SlPHAN can be translated into diverse final leaf morphologies.

Is salK/SalR , a two-component signal transduction system , essential for full virulence of highly invasive Streptococcus suis serotype 2?

Streptococcus suis serotype 2 (S. suis 2, SS2) has evolved into a highly infectious entity, which caused the two recent large-scale outbreaks of human SS2 epidemic in China, and is characterized by a toxic shock-like syndrome. However, the molecular pathogenesis of this new emerging pathogen is still poorly understood. 89K is a newly predicted pathogenicity island (PAI) which is specific to Chinese epidemic strains isolated from these two SS2 outbreaks. Further bioinformatics analysis revealed a unique two-component signal transduction system (TCSTS) located in the candidate 89K PAI, which is orthologous to the SalK/SalR regulatory system of Streptococcus salivarius. Knockout of salKR eliminated the lethality of SS2 in experimental infection of piglets. Functional complementation of salKR into the isogenic mutant DeltasalKR restored its soaring pathogenicity. Colonization experiments showed that the DeltasalKR mutant could not colonize any susceptible tissue of piglets when administered alone. Bactericidal assays demonstrated that resistance of the mutant to polymorphonuclear leukocyte (PMN)-mediated killing was greatly decreased. Expression microarray analysis exhibited a transcription profile alteration of 26 various genes down-regulated in the DeltasalKR mutant.

These findings suggest that SalK/SalR is requisite for the full virulence of ethnic Chinese isolates of highly pathogenic SS2, thus providing experimental evidence for the validity of this bioinformatically predicted PAI.

Is the 2015 Dietary Guidelines for Americans associated with a more nutrient-dense diet and a lower risk of obesity?

Dietary pattern analysis represents a departure from the traditional focus on single foods and nutrients and provides a comprehensive understanding of the role of the diet in chronic disease prevention and etiology. Dietary patterns of Canadians have not been evaluated comprehensively with the use of an updated a priori dietary quality index. We aimed to update the Dietary Guidelines for Americans Adherence Index (DGAI) on the basis of the 2015 Dietary Guidelines for Americans (DGA), to evaluate the construct validity and reliability of the revised index, and to examine whether closer adherence to this index is associated with a lower risk of obesity with or without an accompanying chronic disease. Data from 11,748 participants (≥18 y of age) in the cross-sectional Canadian Community Health Survey cycle 2.2 were used in weighted multivariate analyses. Multinomial logistic regression was used to test the association between diet quality and obesity risk. With the use of principal component analyses, the multidimensionality of the 2015 DGAI was confirmed, and its reliability was shown with a high Cronbach's α = 0.75. Moving from the first to the fourth (healthiest) quartile of the 2015 DGAI score, there was a trend toward decreased energy (2492 ± 26 compared with 2403 ± 22 kcal, respectively; ±SE) and nutrients of concern (e.g., sodium), whereas intakes of beneficial nutrients increased (P-trend < 0.05). In the age- and sex-adjusted model, a lack of adherence to the 2015 DGA recommendations increased the OR of being unhealthy obese from 1.42 (95% CI: 1.02, 1.99) in quartile 3 to 2.08 (95% CI: 1.49, 2.90) in quartile 2 to 2.31 (95% CI: 1.65, 3.23) in the first quartile of the 2015 DGAI score, compared with the fourth quartile (healthiest) (P-trend < 0.0001). The odds of being obese without a chronic disease (healthy obese) and having a chronic disease without being obese also increased in the lowest DGAI quartile compared with the highest DGAI quartile, albeit not as much as in the unhealthy obese group.

The 2015 DGAI provides a valid and reliable measure of diet quality among Canadians.

Do imputing variance estimates alter the conclusions of a meta-analysis with continuous outcomes : a case study of changes in renal function after living kidney donation?

To assess how different imputation methods used to account for missing variance data in primary studies influence tests of heterogeneity and pooled results from a meta-analysis with continuous outcomes. Point and variance estimates for changes in serum creatinine, glomerular filtration rate, systolic blood pressure, and diastolic blood pressure were variably reported among 48 primary longitudinal studies of living kidney donors (71%-78% of point estimates were reported, 8%-13% of variance data were reported). We compared the results of meta-analysis, which either were restricted to available data or used four methods to impute missing variance data. These methods used reported P-values, reported nonparametric summaries, results from other similar studies using multiple imputation, or results from estimated correlation coefficients. Significant heterogeneity was present in all four outcomes regardless of the imputation methods applied. The random effects point estimates and 95% confidence intervals varied little across imputation methods, and the differences were not clinically significant.

Different methods to impute the variance data in the primary studies did not alter the conclusions from this meta-analysis of continuous outcomes. Such reproducibility increases confidence in the results. However, as with most meta-analyses, there was no gold standard of truth, and results must be interpreted judiciously. The generalization of these findings to other meta-analyses, which differ in outcomes, missing data, or between-study heterogeneity, requires further consideration.

Is aberrant expression of trefoil factor 3 associated with colorectal carcinoma metastasis?

Recent evidence has indicated that the trefoil factor family possesses pivotal roles in the progression of human cancer. Aberrant expression of trefoil factor 3 (TFF3) has been reported to correlate with an aggressive tumor phenotype. However, the clinical importance of TFF3 expression in colorectal carcinomas (CRCs) has rarely been addressed. To investigate the putative role of TFF3 in colorectal carcinogenesis and progress, and to clarify whether TFF3 could be a serum marker for CRCs. Fifty-six CRCs were sequenced for TFF3 mutations; subsets of the primary tumors were subjected to real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry analyses and serum TFF3 was detected by enzyme-linked immunosorbent assay (ELISA) for patients with CRCs. No variants were detected in the code area of TFF3; TFF3 mRNA is increased in CRCs but not up to statistic significance when compared with paired normal colonic mucosa; TFF3 staining by immunohistochemistry in primary CRCs showed that increased expression of TFF3 is associated with lymph node metastases(LNM), and no significant differences were found with respect to patient's sex, cancer cell differentiation and stage. Serum TFF3 is significantly elevated in patients with CRCs, especially CRCs with LNM.

The results indicate that TFF3 point mutations seem to be a rare event in colorectal carcinogenesis; TFF3 expression may play a role in promoting lymph node metastases of CRCs and serum TFF3 may be a potential useful marker for patients with CRCs and their metastases.

Is endoglin modulation of TGF-beta1-induced collagen synthesis dependent on ERK1/2 MAPK activation?

Transforming growth factor-beta1 (TGF-beta1) plays a pivotal role in the extracellular matrix accumulation observed in fibrotic diseases. Endoglin is an important component of the TGF-beta receptor complex highly expressed in tissues undergoing fibrotic processes. Endoglin expression regulates the effect of TGF-beta on extracellular matrix synthesis. The purpose of our study has been to understand the molecular mechanism by which endoglin exerts its effects on fibrosis and the possible role of MAP kinases in these effects. We have assessed in mock and in endoglin-transfected L6E9 myoblasts the effect of TGF-beta1 on collagen mRNA by Northern blot and effect of TGF-beta1 on collagen content in the cultured medium by [(3)H]-Proline incorporation into collagen proteins. Total and activated MAPK and their role on collagen synthesis were assessed by Western blot. TGF-beta1 induced an increase on alpha(2) (I) collagen mRNA expression and collagen accumulation in mock-transfected myoblasts, whereas the response was much lower in endoglintransfected cells. TGF-beta1 activated the ERK1/2 and p38 MAPK pathways but not the JNK pathway in L6E9 myoblasts. TGF-beta1-induced alpha(2) (I) collagen mRNA expression and collagen accumulation were completely inhibited by SB203580, in either mock or endoglintransfected myoblasts. PD98059 increased TGF-beta1 induced-collagen synthesis and accumulation in endoglin-transfected myoblasts but not in mock cells.

Our studies demonstrate that TGF-beta1- induced collagen synthesis is mediated by p38 MAPK activation in L6E9 myoblasts. Furthermore, endoglin expression reduces basal and TGF-beta1 induced collagen synthesis when ERK1/2 pathway is operating.

Does lower sternal reinforcement improve the stability of sternal closure?

This study uses a mechanical testing system to evaluate three methods of sternal closure. Twelve sternal replicas composed of a polyurethane foam bone analogue were divided in the midline and reapproximated using three stainless steel wire techniques: six simple wires (6S), six figure-of-eight wires (6F8), or seven simple wires (7S), which included an extra wire at the lower sternum. The closures were subjected to increasing lateral distraction from 0 to 400 Newtons (N) (1 N = 0.224 lbs), and motion was measured using transducers stationed across the manubrium, midsternum, and lower sternum. With each method of closure, the manubrium was the most stable, the lower sternum the least stable, and the midsternum intermediate between the other two. There were also differences between sternal closure methods, but only at the lower sternum. Less sternal distraction was measured with the 7S than the 6S and 6F8 methods, starting at 100 N (0.20 +/- 0.06 mm vs 0.48 +/- 0.19 and 0.39 +/- 0.10, p = 0.003), and progressively increasing until the study was stopped at 400 N (1.64 +/- 0.39 mm vs 4.92 +/- 1.73 and 5.1 +/- 1.43 mm, p = 0.003).

These data show that the lower sternum is the site of greatest instability and that reinforcement of this area with an additional wire effectively stabilizes the closure. Figure-of-eight wires are not superior to simple wires.

Antenatal bowel dilatation in gastroschisis: a bad sign?

Antenatal ultrasound scans and subsequent postnatal notes were reviewed for cases of isolated gastroschisis from 2004 to 2008. Bowel dilatation was defined as>10 mm diameter. The type (intra- and/or extra-abdominal dilatation), maximum bowel diameter and gestation at which bowel dilatation was first seen was recorded. Outcome measures included number of days of TPN, type of closure (primary or delayed) and postnatal complications. There were 60 antenatal cases of gastroschisis over 5 years. Postnatal notes of 47 were obtained. 38 (81%) had antenatal bowel dilatation, 9 did not. Of those with bowel dilatation, 24 underwent primary closure, 9 required silos, 2 a patch and 3 stomas. Seven (15%) had bowel atresia or necrosis. Three died: NEC, septicaemia and CMV hepatitis. The average time on TPN for those without major complications was 22 days. Of those with no bowel dilatation, 4 had primary closure, 3 a silo, 1 a patch and 1 a stoma. Two had complications requiring further surgery, including one atresia resulting in short gut syndrome. There were no deaths. The mean number of days on TPN was 23. The results also demonstrated no significant correlation between gestation when dilatation was first seen, or degree of dilatation, or both these factors combined, and time of TPN requirement.

These results do not support the current practice of using antenatal bowel dilatation as a prognostic indicator.

Does targeting NADPH oxidase decrease oxidative stress in the transgenic sickle cell mouse penis?

Sickle cell disease (SCD) is a state of chronic vasculopathy characterized by endothelial dysfunction and increased oxidative stress, but the sources and mechanisms responsible for reactive oxygen species (ROS) production in the penis are unknown. We evaluated whether SCD activates NADPH oxidase, induces endothelial nitric oxide synthase (eNOS) uncoupling, and decreases antioxidants in the SCD mouse penis. We further tested the hypothesis that targeting NADPH oxidase decreases oxidative stress in the SCD mouse penis. SCD transgenic (sickle) mice were used as an animal model of SCD. Hemizygous (hemi) mice served as controls. Mice received an NADPH oxidase inhibitor apocynin (10 mM in drinking water) or vehicle. Penes were excised at baseline for molecular studies. Markers of oxidative stress (4-hydroxy-2-nonenal [HNE]), sources of ROS (eNOS uncoupling and NADPH oxidase subunits p67(phox) , p47(phox) , and gp91(phox) ), and enzymatic antioxidants (superoxide dismutase [SOD]1, SOD2, catalase, and glutathione peroxidase-1 [GPx1]) were measured by Western blot in penes. Sources of ROS, oxidative stress, and enzymatic antioxidants in the SCD penis. Relative to hemi mice, SCD increased (P<0.05) protein expression of NADPH oxidase subunits p67(phox) , p47(phox) , and gp91(phox) , 4-HNE-modified proteins, induced eNOS uncoupling, and reduced Gpx1 expression in the penis. Apocynin treatment of sickle mice reversed (P<0.05) the abnormalities in protein expressions of p47(phox) , gp91(phox) (but not p67(phox) ) and 4-HNE, but only slightly (P>0.05) prevented eNOS uncoupling in the penis. Apocynin treatment of hemi mice did not affect any of these parameters.

NADPH oxidase and eNOS uncoupling are sources of oxidative stress in the SCD penis; decreased GPx1 further contributes to oxidative stress. Inhibition of NADPH oxidase upregulation decreases oxidative stress, implying a major role for NADPH oxidase as a ROS source and a potential target for improving vascular function in the SCD mouse penis.

Does desipramine attenuate working memory impairments induced by partial loss of catecholamines in the rat medial prefrontal cortex?

The density of tyrosine hydroxylase-immunoreactive (TH-IR) axons in the prefrontal cortex of schizophrenic subjects may be reduced by as much as 50% in the deep cortical layers (Am J Psychiatry 156:1580-1589, 1999). Previously, we demonstrated that approximately 60% loss of TH-IR axons in the rat medial prefrontal cortex (mPFC) decreases local basal and stress-evoked extracellular dopamine (DA) concentrations, suggesting that moderate loss of DA axons in the mPFC is sufficient to alter the neurochemical activity of the remaining DA neurons (Neuroscience 93:497-505, 1999). To further assess the functional consequences of partial mPFC DA depletion, we examined the effects of 6-hydroxydopamine lesions of the rat mPFC on behavior in a T-maze delayed-response task. We also assessed whether chronic administration of the norepinephrine (NE) uptake inhibitor, desipramine (DMI), attenuates lesion-induced deficits in T-maze performance. Previous research indicates that inhibition of NE transport in the mPFC results in a concomitant increase in extracellular DA and NE. Moderate loss of mPFC DA and NE (approximately 50 and 10% loss, respectively) was sufficient to impair delayed-response behavior, in part due to an increase in perseverative responding. Chronic DMI treatment (3 mg/kg delivered via osmotic pumps) impaired performance of control rats but attenuated the deficits in delayed-response behavior in rats previously sustaining loss of mPFC DA and NE (approximately 75 and 35% loss, respectively).

These data suggest that moderate loss of DA and NE in the prefrontal cortex is sufficient to impair cognitive function, and these behavioral effects are attenuated by inhibition of the NE transporter.

Are mouse mammary tumor virus-like gene sequences present in lung patient specimens?

Previous studies have reported on the presence of Murine Mammary Tumor Virus (MMTV)-like gene sequences in human cancer tissue specimens. Here, we search for MMTV-like gene sequences in lung diseases including carcinomas specimens from a Mexican population. This study was based on our previous study reporting that the INER51 lung cancer cell line, from a pleural effusion of a Mexican patient, contains MMTV-like env gene sequences. The MMTV-like env gene sequences have been detected in three out of 18 specimens studied, by PCR using a specific set of MMTV-like primers. The three identified MMTV-like gene sequences, which were assigned as INER6, HZ101, and HZ14, were 99%, 98%, and 97% homologous, respectively, as compared to GenBank sequence accession number AY161347. The INER6 and HZ-101 samples were isolated from lung cancer specimens, and the HZ-14 was isolated from an acute inflammatory lung infiltrate sample. Two of the env sequences exhibited disruption of the reading frame due to mutations.

In summary, we identified the presence of MMTV-like gene sequences in 2 out of 11 (18%) of the lung carcinomas and 1 out of 7 (14%) of acute inflamatory lung infiltrate specimens studied of a Mexican Population.

Does vitamin D modulate peripheral immunity in patients with Behçet 's disease?

There is little knowledge about clinical and immunological variables associated with vitamin D insufficiency in inflammatory diseases. We sought to investigate disease variables associated with vitamin D levels in patients with Behçet's disease (BD) and its interaction with inflammatory responses. One hundred and sixty BD patients (102 patients in active stage) were enrolled in a study assessing the relationship between serum vitamin D concentrations and disease activity. As control diseases we studied 22 Rheumatoid arthritis (RA) and 30 multiple sclerosis (MS) patients. Serum concentrations of vitamin D were assayed with a radioimmunoassay kit. To assess the correlation between inflammatory mediators, immune cell expression and vitamin D, 20 active BD patients and 18 healthy controls were investigated: T-cell subsets and Treg cells were quantified by flow cytometry. Th1/Th2 ratio and Th17 were studied by intracytoplasmic cytokines expression (IFN-γ, IL-4, IL-10 and IL-17). Decreased levels of vitamin D were observed in active BD patients compared to patients in the inactive stage and to healthy controls (p=0.0246; p=0.0001). A low significant difference was observed between inactive BD and healthy controls (p=0.004). Active BD expressed higher vitamin D levels than RA (p=0.007) and MS (p=0.044) patients (p=0.0238). In active BD, vitamin D levels were correlated with CRP and ESR. Serum levels of vitamin D correlated positively with the number of Treg cells (r=0.640; p=0.0024). The IFN-γ/IL-4 ratio (Th1/Th2) was inversely correlated with serum 25(OH)D levels (r=-0.599; p=0.0053).

Active BD was associated with lower serum Vitamin D levels. Our results showed that low levels of vitamin D were associated with a decrease in Treg cells and a skewing of the Th1/Th2 balance towards Th1. These findings suggest that vitamin D is an important promoter of T cell regulation in vivo in BD patients. As suggested in other inflammatory/autoimmune diseases, vitamin D may modulate inflammatory mediators.

Does statin and Metformin Use prolong Survival in Patients With Resectable Pancreatic Cancer?

The aim of this study was to investigate the impact of statin and metformin therapy on disease outcome for patients with pancreatic ductal adenocarcinoma (PDAC). This retrospective study included 171 PDAC patients who underwent surgical resection at the Stanford Cancer Institute between 1998 and 2013. No patients received neoadjuvant therapy. Statin and metformin use was defined as use during initial consult and continuing upon discharge from the hospital after surgery. Dose of each medication was recorded, as was the type of statin taken. The median follow-up for all patients was 11.23 months (range, 0.2-105.0 months). Among the 171 patients included in our analysis, 18 patients (10.5%) took metformin and 34 patients (19.9%) took statins. Statin use was associated with better overall survival (OS) in patients with PDAC (P = 0.011). Metformin use was also associated with better OS (P = 0.035). The use of statins remained significant on multivariate analysis for OS (P = 0.014; hazards ratio, 0.33; 95% confidence interval, 0.139-0.799), but metformin use did not (P = 0.33; hazards ratio 0.60, 95% confidence interval, 0.211-1.675).

Statin and metformin use is associated with improved OS in patients with resectable PDAC. These medications should be further investigated for possible long-term use in the general population.

Is the rapidly expanding CRF01_AE epidemic in China driven by multiple lineages of HIV-1 viruses introduced in the 1990s?

We sought to comprehensively analyze the origin, transmission patterns and sub-epidemic clusters of the HIV-1 CRF01_AE strains in China. Available HIV-1 CRF01_AE samples indentified in national molecular epidemiologic surveys were used to generate near full-length genome (NFLG) sequences. The new and globally available CRF01_AE NFLG sequences were subjected to phylogenetic and Bayesian molecular clock analyses, and combined with epidemiologic data to elucidate the history of CRF01_AE transmission in China. We generated 75 new CRF01_AE NFLG sequences from various risk populations covering all major CRF01_AE epidemic regions in China. Seven distinct phylogenetic clusters of CRF01_AE were identified. Clusters 1, 2 and 3 were prevalent among heterosexuals and IDUs in southern and southwestern provinces. Clusters 4 and 5 were found primarily among MSM in major northern cities. Clusters 6 and 7 were only detected among heterosexuals in two southeast and southwest provinces. Molecular clock analysis indicated that all CRF01_AE clusters were introduced from Southeast Asia in the 1990s, coinciding with the peak of Thailand's HIV epidemic and the initiation of China's free overseas travel policy for their citizens, which started with Thailand as the first destination country.

China's HIV-1 epidemic of sexual transmissions, was initiated by multilineages of CRF01_AE strains, in contrast to the mono-lineage epidemic of B' strain in former plasma donors and IDUs. Our study underscores the difficulty in controlling HIV-1 sexual transmission compared with parenteral transmission.

Does trichosanthes kirilowii tuber extract induce G2/M phase arrest via inhibition of tubulin polymerization in HepG2 cells?

Trichosanthes kirilowii tuber is one of most popular herbal plant of East Asia, which has been prescribed for patients with diabetes, rigorous coughing, breast abscesses, and cancer-related symptoms. To investigated the anticancer properties of the methanol extract of Trichosanthes kirilowii tuber (TKE), focusing on cell cycle arrest and microtubule instability in HepG2 cells. Cell growth and death were checked using a CCK-8 assay and a LDH release assay respectively. Cell cycle was analyzed by FACS after PI staining. Immunofluorescence, Western blot, real-time PCR for tubulin were performed. TKE treatment inhibited cell growth at around 25 microg/mL of IC50 in a CCK-8 assay and a LDH release assay, but did not result in cell death. We found that TKE induced cell cycle arrest at the G2/M phase in a time-dependent manner. However, an immunofluorescence assessment of beta-tubulin revealed a dramatically reduced amount of polymerized tubulin after TKE treatment. Furthermore, TKE treatment radically decreased the polymerized portion of soluble tubulin in a dose-dependent manner, as did colchicine; the effects, however, were opposite to those of paclitaxel in comparative analysis of polymerized to soluble tubulin. We also found that TKE treatment moderately affected alpha-tubulin protein production, but not that of beta-tubulin and its gene expression using a Western assay and real-time PCR.

Anticancer mechanisms of TKE linked to the inhibition of tubulin polymerization, through which it exerts cell cycle arrest at the G2/M phase in the HepG2 cell line.

Does genome-wide survey indicate involvement of loci on canine chromosomes 7 and 31 in patellar luxation in Flat-Coated Retrievers?

Patellar luxation is an orthopedic disorder in which the patella moves out of its normal location within the femoral trochlea of the knee and it can lead to osteoarthritis, lameness, and pain. In dogs it is a heritable trait, with both environmental and genetic factors contributing to the phenotype. The prevalence of patellar luxation in the Dutch Flat-Coated Retriever population is 24%. In this study, we investigated the molecular genetics of the disorder in this population. Genome-wide association analysis of 15,823 single nucleotide polymorphisms (SNPs) in 45 cases and 40 controls revealed that patellar luxation was significantly associated with a region on chromosome CFA07, and possibly with regions on CFA03, CFA31, and CFA36. The exons of the genes in these regions, 0.5 Mb combined, were analyzed further. These exons from 15 cases and a pooled sample from 15 controls were enriched using custom genomic hybridization arrays and analyzed by massive parallel DNA sequencing. In total 7257 variations were detected. Subsequently, a selection of 144 of these SNPs were genotyped in 95 Flat-Coated Retrievers. Nine SNPs, in eight genes on CFA07 and CFA31, were associated with patellar luxation (P <10-4). Genotyping of these SNPs in samples from a variety of breeds revealed that the disease-associated allele of one synonymous SNP in a pseudogene of FMO6 was unique to Flat-Coated Retrievers.

Genome-wide association analysis followed by targeted DNA sequencing identified loci on chromosomes 7 and 31 as being involved in patellar luxation in the Flat-Coated Retriever breed.

Does recombinant human Flt3 ligand exert both direct and indirect effects on hematopoiesis?

To investigate the direct effects of the Flt3 ligand (FL) on hematopoiesis, such as the stimulation of the formation of hematopoietic colonies and the proliferation of dendritic cells, as well as the indirect stimulation of hematopoiesis, especially via the proliferation of endothelial cells. Mononuclear cells from human cord blood were plated in methylcellulose medium containing different cytokines to induce hematopoietic colony formation. Dendritic cells (DCs) were induced from the mononuclear cells with a cytokine cocktail with or without recombinant human soluble FL (rhFL; 100 ng/ml). The Flt3 receptors on the surface of a human microvascular endothelial cell line (ECV) were analyzed by flow cytometry. The proliferation of ECV stimulated by rhFL was measured with the microculture tetrazolium assay. The levels of FL, IL-6, IL-8, G-CSF and GM-CSF in the supernatant of ECV cultures were measured by enzyme linked immunoabsorbent assay (ELISA). rhFL stimulates colony formation from cord blood when used as a sole stimulant. FL in combination with other cytokines increased colony formation significantly. The number of DCs was approximately 2.5 times higher when rhFL was used. rhFL stimulates the proliferation of ECV on which Flt3 receptors are expressed. Furthermore, ECV secretes FL, IL-6, IL-8, G-CSF and GM-CSF, which were augmented by tumor necrosis factor-alpha and rhFL.

rhFL enhances hematopoietic colony formation and DC proliferation from human cord blood cells. FL not only stimulates the proliferation of ECV, but is also secreted by ECV. FL may exert direct and indirect effects on hematopoiesis.

Does [ Amniotic membrane transplantation improve herpetic keratitis by local and not by systemic effects ]?

The immune mediated HSV-1 stromal keratitis (HSK) rapidly improves after amniotic membrane transplantation (AMT). This study investigated whether AMT modulates the T cell response and whether the anti-inflammatory action of AMT is due to local or systemic effects. Corneas of BALB/c mice were infected with 10(5) (PFU) of HSV-1. Animals with ulcerating keratitis on day 14 post-infection were divided into 4 groups: group 1 ( n=12): right eye AMT;group 2 ( n=12): right eye tarsorrhaphy; group 3 ( n=8): right eye tarsorrhaphy, left eye AMT;group 4 ( n=8): both eyes tarsorrhaphy. The mice were examined for clinical signs of HSV keratitis after 2 days. Corneal sections were studied histologically and the inflammatory cell infiltration was studied by immunohistochemical staining. DTH response and the HSV-specific 3H-thymidin-uptake were compared between the groups. Compared to group 2, ulceration and stromal inflammation was profoundly improved in group 1 ( p<0.01). The corneas in the AMT mice had fewer inflammatory cells, CD3+,CD4+ and CD8+ cells than the control mice ( p<0.01). There were no significant differences between groups 1 and 2 with respect to the delayed type hypersensitivity reaction (DTH response) and the HSV-specific 3H-thymidin uptake. AMT or tarsorrhaphy on the left eyes in groups 3 and 4 had no influence on the course of keratitis or the T cell response.

Ulcerating herpetic keratitis markedly improves after AMT. Our observations indicate that this is caused predominantly by local and not by systemic AMT-related effects.

Does methylamine-dependent release of nitric oxide and dopamine in the CNS modulate food intake in fasting rats?

Methylamine is an endogenous aliphatic amine exhibiting anorexigenic properties in mice. The aim of this work was to show whether methylamine also modifies feeding behaviour in rats and, if so, to identify the mediator(s) responsible for such effects. Microdialysis experiments with the probe inserted in the periventricular hypothalamic nucleus were carried out in 12 h starved, freely moving rats. Collected perfusate samples following methylamine injection (i.c.v.) were analysed for nitric oxide by chemiluminescence and for dopamine and 5-hydroxytryptamine content by HPLC. Kv1.6 potassium channel expression was reduced by antisense strategy and this decrease quantified by semi-quantitative RT-PCR analysis. Methylamine showed biphasic dose-related effects on rat feeding. At doses of 15-30 microg per rat, it was hyperphagic whereas higher doses (60-80 microg) were hypophagic. Methylamine stimulated central nitric oxide (+115% vs. basal) following hyperphagic and dopamine release (60% over basal values) at hypophagic doses, respectively. Treatment with L-N(G)-nitro-L-arginine-methyl ester (i.c.v. 2 microg 10 microl(-1)) or with alpha-methyl-p-tyrosine (i.p. 100 mg kg(-1)) before methylamine injection, reduced nitric oxide output and hyperphagia, or dopamine release and hypophagia respectively. Moreover, hypophagia and hyperphagia, as well as nitric oxide and dopamine release were significantly reduced by down-regulating brain Kv1.6 potassium channel expression.

The effects of methylamine on feeding depend on the hypothalamic release of nitric oxide and dopamine as a result of interaction at the Kv1.6 channels. The study of methylamine levels in the CNS may provide new perspectives on the physiopathology of alimentary behaviour.

Is primary tumor volume an independent predictor of outcome within pT4a-staged tongue carcinoma?

We evaluated the heterogeneity of primary tumor volume (PTV) within tumors of the same pT4a-staged tongue carcinoma and to elucidate the effects of PTV on treatment outcomes in patients with pT4a-staged tongue carcinoma. Fifty-eight patients with newly diagnosed pT4a-staged tongue carcinoma who received surgery were enrolled onto this study. Magnetic resonance imaging-derived PTV was measured by the summation-of-area technique. The mean PTV was 24.55 ml, with a range of 5.32 to 119.64 ml. The receiver operating characteristic curve was applied, and the optimal cutoff volume was 23 ml. Large PTV was associated with a significantly poor disease-specific survival (P = 0.010) by the log rank test. The Cox regression model also revealed that large PTV (P = 0.026) and positive lymphatic node metastasis (P = 0.004) were statistically significant in the prognosis of T4a-staged tongue carcinoma.

A substantial variation of PTV was present within the same pT4a-staged tongue carcinoma, and PTV represented an important prognostic factor. In the light of these findings, we suggest that taking the PTV into account in pT4a-staged tongue carcinoma would better refine the newest revised T classification, and the treatment strategies may be different.

Is miRNA-140 a negative feedback regulator of MMP-13 in IL-1β-stimulated human articular chondrocyte C28/I2 cells?

Osteoarthritis is a degenerative joint disease, in which matrix metalloproteinase (MMP)-13 plays an important role. This study aimed to investigate miRNA-140-mediated negative regulation of MMP-13 expression in interleukin-1β (IL-1β)-stimulated human cartilage cells. The human cartilage cell line C28/I2 was cultured in the presence of IL-1β to mimic an osteoarthritic environment. Expression of miRNA-140 and MMP-13 was analyzed after 48 h by real-time RT-PCR and western blot analyses. MiRNA-140 mediated regulation of MMP-13 expression was analyzed by luciferase reporter assays and anti-miRNA-140 oligonucleotide transfection. Furthermore, miRNA-140 and MMP-13 expression was analyzed following DHMEQ treatment. Expression of miRNA-140 and MMP-13 was elevated in IL-1β-stimulated C28/I2 cells. Bioinformatic prediction showed that the 3'-UTR of MMP-13 mRNA contained a potential binding miRNA-140 site and luciferase mRNA fused with 3'-UTR of MMP-13 mRNA was shown to be repressed by miRNA-140 in reporter assays. Expression of MMP-13 was elevated in IL-1β-stimulated C28/I2 cells following anti-miRNA-140 oligonucleotide transfection. NF-κB activity was inhibited in DHMEQ treated IL-1β-stimulated C28/I2 cells and was associated with decreased miRNA-140 and MMP-13 expression.

Expression of miRNA-140 and MMP-13 was induced by IL-1β. Expression of miRNA-140 inhibited MMP-13 in C28/I2 cells. Expression of miRNA-140 and MMP-13 was shown to be NF-κB-dependent.

Does moxibustion regulate inflammatory mediators and colonic mucosal barrier in ulcerative colitis rats?

To observe the efficacy and mechanism of grain-sized moxibustion at different acupoints in a rat model of ulcerative colitis (UC). Sprague-Dawley rats were randomly divided into control, UC model, grain-sized moxibustion at a single acupoint (CV 12), grain-sized moxibustion at two acupoints (CV 12 and CV 4), grain-sized moxibustion at three acupoints (CV 12, CV 4, and ST 36), and medication groups (n = 8/group). The UC model was established by enema of trinitrobenzene sulfonic acid. Direct moxibustion was used once a day for 7 d. Disease activity index (DAI) was evaluated before and after the treatment. Morphologic changes of intestinal tissue were observed under an optical microscope. The expression of tumor necrosis factor (TNF)-α and p38 mitogen-activated protein kinase (p38MAPK) in colonic tissue was detected using Western blot, and the levels of occludin and zonula occludens-1 (ZO-1) mRNAs were detected using reverse transcription PCR. Compared with the control group, the intestinal mucosae were incomplete in the model group, glandular structures were irregular, and submucosae were edematous, hyperemic, and infiltrated with inflammatory cells. The DAI scores and expression of TNF-α and p38MAPK were increased significantly in the model group compared to controls (Ps < 0.01), while the mRNA levels of occludin and ZO-1 were reduced significantly (Ps < 0.01). Compared with the model group, colonic mucosa and the arrangement of glands were complete and regular in the treatment groups. DAI scores and the expression of TNF-α and p38MAPK were reduced significantly in moxibustion groups compared to controls (Ps < 0.01), while the mRNA levels of occludin and ZO-1 were increased significantly (Ps < 0.01). The improvements in the above indices in the three acupoints group and the medication group were superior to those in the single and two acupoints groups (all P < 0.05).

Reduction of TNF-α and p38MAPK and increased expression of occludin and ZO-1 in colonic tissue represent a potential mechanism for improved intestinal mucosal tissue repair with grain-sized moxibustion.

Are bDNF SNPs implicated in comorbid alcohol dependence in schizophrenia but not in alcohol-dependent patients without schizophrenia?

The functional BDNF single nucleotide polymorphism (SNP) rs6265 has been associated with many disorders including schizophrenia and alcohol dependence. However, studies have been inconsistent, reporting both positive and negative associations. Comorbid alcohol dependence has a high prevalence in schizophrenia so we investigated the role of rs6265 in alcohol dependence in Australian populations of schizophrenia and alcohol-dependent patients. Two BDNF SNPs rs6265 and a nearby SNP rs7103411 were genotyped in a total of 848 individuals. These included a schizophrenia group (n = 157) and a second schizophrenia replication group (n = 235), an alcohol-dependent group (n = 231) that had no schizophrenia diagnosis and a group of healthy controls (n = 225). Allelic association between rs7103411 and comorbid alcohol dependence was identified (P = 0.044) in the primary schizophrenia sample. In the replication study, we were able to detect allelic associations between both BDNF SNPs and comorbid alcohol dependence (rs6265, P = 0.006; rs7103411, P = 0.014). Moreover, we detected association between both SNPs and risk-taking behaviour after drinking (rs6265, P = 0.005; rs7103411, P = 0.009) and we detected strong association between both SNPs and alcohol dependence in males (rs6265, P = 0.009; rs7103411, P = 0.013) while females showed association with multiple behavioural measures reflecting repetitive alcohol consumption. Haplotype analysis revealed the rs6265-rs7103411 A/C haplotype is associated with comorbid alcohol dependence (P = 0.002). When these SNPs were tested in the non-schizophrenia alcohol-dependent group we were unable to detect association.

We conclude that these BDNF SNPs play a role in development of comorbid alcohol dependence in schizophrenia while our data do not indicate that they play a role in alcohol-dependent patients who do not have schizophrenia.

Is glomerular expression of nephrin and synaptopodin , but not podocin , decreased in kidney sections from women with preeclampsia?

Preeclampsia is a pregnancy-specific disorder characterized by hypertension and proteinuria. In other disease states, proteinuria has been linked to altered expressions of podocyte foot-process proteins, but this has not been studied in women with preeclampsia. We sought to test the hypothesis that proteinuria in preeclampsia is associated with dysregulated expression of the podocyte cytoskeleton and/or tight junction proteins. Renal tissue was obtained from autopsy material from seven women who had severe preeclampsia during the second half of their pregnancies up to 48 h after delivery, and who subsequently died. As controls, we used autopsy material from two women who died accidentally during the second half of their otherwise normal pregnancies. Immunohistochemical stains for nephrin, synaptopodin and podocin were performed on representative sections prepared from paraffin-embedded material. Expression of both nephrin and synaptopodin was markedly decreased in preeclamptic compared with control kidney sections. By contrast, both cases and controls demonstrated strong staining for podocin.

We conclude that down-regulation of nephrin and synaptopodin is associated with proteinuria in women with preeclampsia. Recent studies have demonstrated that soluble vascular endothelial growth factor receptor 1 (sFlt-1) levels are elevated in preeclampsia compared with normal pregnancy. Studies in mice have shown that sFlt-1 may play a role in inducing proteinuria by neutralizing vascular endothelial growth factor (VEGF) and suppressing nephrin. Proteinuria and elevations of sFlt-1 in preeclampsia are temporally related, further supporting a possible role of sFlt-1 in the dysregulation of podocyte foot-process proteins.

Is hyperresponsiveness of the mucosal barrier in Crohn 's disease tumor necrosis factor-dependent?

Crohn's disease (CD) is associated with gut barrier dysfunction. Besides the baseline barrier defect, a subgroup of patients also expresses an intestinal barrier hyperresponsiveness to nonsteroidal anti-inflammatory drugs. We studied whether reducing inflammation and restoring gut barrier dysfunction with anti-tumor necrosis factor (TNF) antibody treatment also antagonizes the permeability increase by oral nonsteroidal anti-inflammatory drug intake in patients with CD. Thirty-one healthy control subjects and 25 patients with active CD were studied. The 31 controls performed intestinal permeability testing for Cr-EDTA before (baseline) and after oral intake of indomethacin (50 + 75 mg). Twenty-five patients carried out a baseline and indomethacin-mediated permeability test before infliximab infusion. The patients repeated either the indomethacin test (12/25) or baseline and indomethacin tests (13/25), 1 month after this treatment. Intestinal permeability was studied by measurement of urinary excretion of Cr-EDTA after oral intake. Increased whole gut permeation before treatment (3.16%; interquartile range [IQR], 2.92-5.72) was restored to normal values (2.47%; IQR, 1.97-2.78) by anti-TNF treatment. Indomethacin increased whole gut permeability significantly more in patients with CD (before anti-TNF: 6.50%; IQR, 4.84-10.38; after anti-TNF: 5.50%; IQR, 3.97-10.09) compared with the healthy subjects (4.66%; IQR, 3.51-5.64). Eleven of 25 patients (44%) had an abnormal whole gut permeability response to indomethacin before anti-TNF, and 9 of them remained hyperresponsive after infusion, despite clinical remission.

Although anti-TNF treatment suppresses inflammation and restores gut barrier function in patients with CD, it does not antagonize the barrier hyperresponsiveness to indomethacin. These data support the notion of an underlying intestinal mucosal barrier hyperresponsiveness in a subset of patients with CD, independent of inflammation.

Does a calcimimetic agent lower plasma parathyroid hormone levels in patients with secondary hyperparathyroidism?

The calcimimetic agent R-568 lowers plasma parathyroid hormone (PTH) levels in hemodialysis patients with mild secondary hyperparathyroidism, but its efficacy in those with more severe secondary hyperparathyroidism has not been studied. Twenty-one patients undergoing hemodialysis three times per week with plasma PTH levels between 300 and 1200 pg/mL were randomly assigned to 15 days of treatment with either 100 mg of R-568 (N = 16) or placebo (N = 5). Plasma PTH and blood ionized calcium levels were measured at intervals of up to 24 hours after oral doses on days 1, 2, 3, 5, 8, 11, 12, and 15. Pretreatment PTH levels were 599 +/- 105 (mean +/- SE) and 600 +/- 90 pg/mL in subjects given R-568 or placebo, respectively, and values on the first day of treatment did not change in those given placebo. In contrast, PTH levels fell by 66 +/- 5%, 78 +/- 3%, and 70 +/- 3% at one, two, and four hours, respectively, after initial doses of R-568, remaining below pretreatment values for 24 hours. Blood ionized calcium levels also decreased after the first dose of R-568 but did not change in patients given placebo. Despite lower ionized calcium concentrations on both the second and third days of treatment, predose PTH levels were 422 +/- 70 and 443 +/- 105 pg/mL, respectively, in patients given R-568, and values fell each day by more than 50% two hours after drug administration. Predose PTH levels declined progressively over the first nine days of treatment with R-568 and remained below pretreatment levels for the duration of study. Serum total and blood ionized calcium concentrations decreased from pretreatment levels in patients given R-568, whereas values were unchanged in those given placebo. Blood ionized calcium levels fell below 1.0 mmol/L in 7 of 16 patients receiving R-568; five patients withdrew from study after developing symptoms of hypocalcemia, whereas three completed treatment after the dose of R-568 was reduced.

The calcimimetic R-568 rapidly and markedly lowers plasma PTH levels in patients with secondary hyperparathyroidism caused by end-stage renal disease.

Is cks1 degraded via the ubiquitin-proteasome pathway in a cell cycle-dependent manner?

Recent work has demonstrated the role of cdc kinase subunit 1 (Cks1) in the ubiquitin-proteasome dependent degradation of CDK inhibitor p27Kip1 protein as an essential cofactor for SCFSkp2 ubiquitin ligase. Although over-expression of Cks1 protein as well as it of Skp2 might be associated with tumour progression via p27Kip1 protein degradation, it is unknown how the cellular level of Cks1 is regulated. Here we show that Cks1 protein is degraded via the ubiquitin-proteasome pathway. Degradation of Cks1 protein was markedly inhibited by proteasome inhibitors. In addition, Cks1 protein was modified with polyubiquitin chains both in vivo and in vitro. Furthermore, we found that degradation of Cks1 protein via the ubiquitin-proteasome pathway was facilitated in M phase during the cell cycle.

These observations suggest that the level of expression of Cks1 protein is regulated at not only the transcriptional level but also the post-translational level via the ubiquitin-proteasome pathway in a cell-cycle-dependent manner.

Are central neuropeptide Y receptors involved in 3rd ventricular ghrelin induced alteration of colonic transit time in conscious fed rats?

Feeding related peptides have been shown to be additionally involved in the central autonomic control of gastrointestinal functions. Recent studies have shown that ghrelin, a stomach-derived orexigenic peptide, is involved in the autonomic regulation of GI function besides feeding behavior. Pharmacological evidence indicates that ghrelin effects on food intake are mediated by neuropeptide Y in the central nervous system. In the present study we examine the role of ghrelin in the central autonomic control of GI motility using intracerobroventricular and IP microinjections in a freely moving conscious rat model. Further the hypothesis that a functional relationship between NPY and ghrelin within the CNS exists was addressed. ICV injections of ghrelin (0.03 nmol, 0.3 nmol and 3.0 nmol/5 microl and saline controls) decreased the colonic transit time up to 43%. IP injections of ghrelin (0.3 nmol - 3.0 nmol kg(-1) BW and saline controls) decreased colonic transit time dose related. Central administration of the NPY1 receptor antagonist, BIBP-3226, prior to centrally or peripherally administration of ghrelin antagonized the ghrelin induced stimulation of colonic transit. On the contrary ICV-pretreatment with the NPY2 receptor antagonist, BIIE-0246, failed to modulate the ghrelin induced stimulation of colonic motility.

The results suggest that ghrelin acts in the central nervous system to modulate gastrointestinal motor function utilizing NPY1 receptor dependent mechanisms.

Does vardenafil increase penile rigidity and tumescence in men with erectile dysfunction after a single oral dose?

To evaluate the effect of two doses of vardenafil hydrochloride on penile rigidity and tumescence while determining the pharmacokinetics. Twenty-one patients with erectile dysfunction completed three oral single-dose regimens (placebo, 20 and 40 mg vardenafil) in a randomized, placebo-controlled, 3-way cross-over study. Penile rigidity and tumescence were measured at the base and tip with a Rigiscan for up to 2 h after dosing. The period included three 20-min repeated episodes of visual sexual stimulation. Blood samples were taken periodically up to 24 h after dosing. After 20 and 40 mg vardenafil, the mean duration of >60% rigidity of the base of the penis was greater than after placebo by 42.9 min (95% Cl 29.3-56.4) and by 49.3 min (95% Cl 35.7-62.9), respectively (p<0.001), and greater than after placebo by 34.6 min (95% Cl 22.1-47.1) for both doses at the tip. Additionally, significantly greater rigidity activity units and tumescence activity units were found for both doses compared with placebo (p<0.001). The plasma concentrations of vardenafil increased rapidly, with a median t(max) of about 40 min and a mean t1/2 of 4.4-4.8 h. Relative bioavailability was slightly higher for the 40-mg dose than for the 20-mg dose. The treatments were well tolerated, although slightly more adverse events, primarily headache, flushing and nasal congestion, were seen with the 40-mg dose compared with placebo.

The findings confirm that vardenafil was able to generate stronger erections of longer duration than placebo under conditions of visual sexual stimulation in patients with erectile dysfunction. The pharmacokinetic, pharmacodynamic and tolerability profiles support vardenafil hydrochloride as a strong candidate for further testing as a treatment for erectile dysfunction.

Can armrests improve comfort and task performance in laparoscopic surgery?

The study sought to determine whether the support provided by armrests influenced task quality, task efficiency, and surgeon comfort during laparoscopic surgery. Complex laparoscopic surgery requires precise movements, and usually long execution times and an uncomfortable stance. Discomfort in the shoulders, back, and neck is an established complaint among laparoscopic surgeons and is related to the unnatural postures adopted during laparoscopic interventions. Discomfort, and the associated fatigue, is a contributory factor in the execution of errors. Nineteen subjects completed a bimanual simulated laparoscopic task both with and without the aid of bilateral armrests. The task was completed in both an ideal unstressed posture and an uncomfortable, stressed elevated posture that more closely represents real laparoscopic operating conditions. Task duration was prolonged sufficiently to precipitate muscular fatigue. The participants also completed a visual analogue scale instrument on level of discomfort symptoms experienced in every part of the upper limbs and vertebral spine. Execution errors (task quality) and completion times (task efficiency) were recorded automatically by the laparoscopic simulator. Error rates and discomfort measures were significantly improved when the armrests were used, but there was no significant change in task completion time.

The use of armrests in simulated laparoscopic surgery brings measurable comfort and task performance benefits, which could transfer to actual surgical procedures.

Is the finding of premalignant lesions associated with smoking cessation in chemoprevention study volunteers?

Screening programs for lung cancer may lead to a heightened awareness of the risks of smoking and enhance quitting. The aim of this study was to evaluate whether the participation on a chemoprevention study for premalignant lesions could influence smoking cessation. Two hundred one volunteers, current (n = 188) and former smokers (n = 13) with more than 20 pack years had been screened for the chemoprevention study. One hundred forty-six of the current smokers at time of chemoprevention study screening have been retrospectively interviewed about their smoking behavior > or =1 year after their first contact for the chemoprevention study. Structured questionnaires were used, and interviews were held by telephone. The quitters at the time of these first interviews were contacted again 4 years after the initial interview about their current smoking behavior. Of the 146 smoking volunteers, 83 were diagnosed with premalignant lesions of the bronchial mucosa and participated in the chemoprevention study, and 63 had no premalignant lesions and were not included in that study.The majority of participants were men: 87 (60%). The mean age of the participants was 52 +/- 9 years, and the mean age at which volunteers started smoking was 15 +/- 3. Mean number of pack years was 47 +/- 27. Ten volunteers in the group without premalignant lesions and 19 in the group with premalignant lesions had quit smoking at time of the first interview. The smoking cessation rate of the total study group was 20%.Univariate logistic regression analysis demonstrated that smoking cessation was only significantly associated with male gender. No significant associations were found between smoking cessation and the finding of premalignant lesions, sex, age, level of addiction, educational level, marital condition, history of cancer/pulmonary diseases, age at start smoking, previous attempts to quit smoking, and motivation to quit smoking.Within the group of subjects who had quit smoking at the time of the first interview, 15 of 29 persons who had stopped smoking at the time of the first interview have reported that participation in the bronchoscopy screening and/or the trial has been of major influence on their decision to stop smoking.

A smoking cessation rate of 20% has been found among volunteers for a chemopreventive trial investigating smoking-related premalignant lesions after almost 2 years after initial contact has been found. Volunteers experienced screening and trial participation as having influenced their smoking cessation. Smoking cessation was significantly associated with male gender, whereas the finding of premalignant lesions by bronchoscopy was not.

Can PCI with drug-eluting stents replace coronary artery bypass surgery?

It is generally known and accepted that percutaneous coronary intervention (PCI) has undergone a patient-relevant innovation with the introduction of drug-eluting stents (DES): prospective, randomized, controlled studies with a primary clinical endpoint have shown that DES, especially those releasing sirolimus from a polymer (SES) or those releasing paclitaxel from a polymer (PES), significantly and relevantly reduce the restenosis rate and hence the number of needed reinterventions in the target vessel (target vessel revascularization [TVR]) as compared to bare metal stents (BMS). For this improvement of quality of life, cost-effectiveness analyses comparing DES and BMS in the US and German health care systems have been reported. For the comparison of DES and coronary artery bypass graft surgery (CABG), no economic data regarding the German health care system have been published. The goal of the present study is to provide such an economic analysis investigating the question whether DES can reduce costs as compared to CABG within the German health care system. Based on the clinical data for PCI with a PES for long and complex lesions (TAXUS VI) and for CABG (ARTS I), the comparison was calculated for the time interval of 1 year. The analysis of the PCI group was derived from a TVR of 11% including a CABG rate of 1%; the CABG group analysis was based on a TVR of 3.8% including a PCI rate of 3.1% and a re-CABG rate of 0.7%. At a stent factor of 2.75, the costs per stent were calculated to be 250 Euro for the BMS and 1,500 Euro for the PES. The total costs for 12 months included the follow-up costs for reinterventions. The results were tested for stability according to a sensitivity analysis. Patients' demographics were well comparable between the PCI and the CABG groups. The primary costs, including the reinterventions, were 7,841 Euro for PCI and 12,415 Euro for CABG. The sensitivity analysis revealed that only at more than eight stents per patient and a need for reintervention of ca. 10% did the cost of PCI with PES reach the level of the cost for CABG.

Within the German health care system, in patients with a social insurance, PCI of long and complex lesions with PES significantly reduces costs as compared to CABG to the amount of 4,574 Euro per patient. The cost-effectiveness analyses of currently ongoing prospective, randomized trials (SYNTAX und FREEDOM) will provide more insight into the economic comparison of PCI with DES and CABG.

Is [ P1208fs mutation in the cardiac myosin binding protein C associated with hypertrophic cardiomyopathy in a Chinese pedigree ]?

To identify the potential mutations in a Chinese pedigree with hypertrophic cardiomyopathy (HCM), and to analyze the genotype-phenotype relationship in this pedigree. Clinical history and physical examinations, electrocardiography (ECG), echocardiography (UCG), cardiac magnetic resonance (CMR) data were obtained from 10 members of a three-generation Chinese family with HCM. A total of 96 genes related to hereditary cardiomyopathy were detected by exon and boarding intron analyses in the proband using second-generation sequencing. Mutations identified in the proband were confirmed by bi-directional Sanger sequencing in the rest 9 family members and 300 healthy controls. Three mutations, including MYBPC3-P1208fs, ANK2-H556R and ANK2-P1974H, were identified in this pedigree. MYBPC3-P1208fs gene mutation was detected in 3 family members (proband, his mother and son), while this mutation was not detected in the rest family members. HCM was diagnosed in the proband and his mother by ECG, UCG and CMR. Son of the proband demonstrated early phenotype of HCM: although UCG and CMR were normal, ECG showed sinus bradycardia and paroxysmal supraventricular arrhythmias as well as ST segment changes. The onset age of HCM diagnosis of the proband and his mother was 42 and 50 years old, presented with palpitation and chest pain, and myocardial fibrosis sign in CMR. Furthermore, we found that left ventricular myocardial fibrosis is related to ECG changes (increasing r wave, ST segment change) in the proband and his mother. No HCM phenotype was evidenced in the 7 family members carrying ANK2-H556R and ANK2-P1974H mutations.

Our results show that MYBPC3-P1208fs gene mutation is associated HCM phenotype in this Chinses pedigree. This mutation is associated with myocardial fibrosis and ST changes in HCM phenotype in this pedigree while ANK2-H556R and ANK2-P1974H mutations are not related to HCM phenotype in this family.

Does clinical evidence support ICD-9-CM diagnosis coding of complications?

Hospital discharge diagnoses, coded by use of the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM), increasingly determine reimbursement and support quality monitoring. Prior studies of coding validity have investigated whether coding guidelines were met, not whether the clinical condition was actually present. To determine whether clinical evidence in medical records confirms selected ICD-9-CM discharge diagnoses coded by hospitals. Retrospective record review of 485 randomly sampled 1994 hospitalizations of elderly Medicare beneficiaries in Califomia and Connecticut. Proportion of patients with specified ICD-9-CM codes representing potential complications who had clinical evidence confirming the coded condition. Clinical evidence supported most postoperative acute myocardial infarction diagnoses, but fewer than 60% of other diagnoses had confirmatory clinical evidence by explicit clinical criteria; 30% of medical and 19% of surgical patients lacked objective confirmatory evidence in the medical record. Across 11 surgical and 2 medical complications, objective clinical criteria or physicians' notes supported the coded diagnosis in>90% of patients for 2 complications, 80% to 90% of patients for 4 complications, 70% to<80% of patients for 5 complications, and<70% for 2 complications. For some complications (postoperative pneumonia, aspiration pneumonia, and hemorrhage or hematoma), a large fraction of patients had only a physician's note reporting the complication.

Our findings raise questions about whether the clinical conditions represented by ICD-9-CM codes used by the Complications Screening Program were in fact always present. These findings highlight concerns about the clinical validity of using ICD-9-CM codes for quality monitoring.

Is severe hypoglycemia associated with antidiabetic oral treatment compared with insulin analogs in nursing home patients with type 2 diabetes and dementia : results from the DIMORA study?

Severe hypoglycemia is associated with cognitive decline and dementia in older persons with type 2 diabetes. The role of antidiabetic treatments on severe hypoglycemia is unknown in dementia. The aims were to determine the prevalence of severe hypoglycemic events and investigate associations among severe hypoglycemic and specific antidiabetic treatments (classes of oral agents and types of insulin analogs) in a large sample of nursing home patients with diabetes according to dementia status. Cross-sectional observational study. A total of 150 nursing homes across Italy. A total of 2258 patients with type 2 diabetes (dementia = 1138, no dementia = 1120). Diagnosis of dementia before nursing home admission. Data were collected regarding functional status, glycemic control, antidiabetic treatments, comorbidities, and biochemical and clinical measurements. Logistic regression models with severe hypoglycemia as the dependent variable were used to test associations with antidiabetic agents. Patients had a mean age (SD) of 82 (8) years, body mass index (BMI) of 25.4 (4.8), fasting plasma glucose (FPG) of 7.5 (3.0) mmol/L, postprandial glucose (PPG) of 10.3 (3.6) mmol/L, HbA1c of 7.1% (54 mmol/L), and impairments in activities of daily living (ADLs) of 3.7 (2.1). Severe hypoglycemia was more prevalent in patients with dementia (18%) compared with patients without dementia (8%). Patients with dementia were older, showed greater ADL impairments, greater number of comorbidities, lower FPG, and higher PPG compared with those without dementia. Adjusted logistic regression models in patients with dementia showed that rapid- and long-acting insulin analogs were associated with reduced odds ratio (OR) (OR 0.333; 95% confidence interval [CI] 0.184-0.602; OR 0.248, 95% CI 0.070-0.882, respectively), whereas sulphonylureas and combined metformin + sulphonylurea were associated with increased ORs (OR 8.805, 95% CI 4.260-18.201; OR 6.639; 95% CI 3.273-14.710, respectively) of experiencing severe hypoglycemia. No correlations were found in patients without dementia.

In older nursing home patients with type 2 diabetes, severe hypoglycemia was significantly higher in dementia. Our findings suggest that sulphonylureas should be used with caution, whereas rapid- and long-acting insulin analogs seem safer.

Does tGF-β induce Up-Regulation of Chondroitin Sulfate Synthase 1 ( CHSY1 ) in Nucleus Pulposus Cells Through MAPK Signaling?

Chondroitin sulfate synthase 1 (CHSY1) is a glycosyltransferases involved in the biosynthesis of chondroitin and dermatan sulfate glycosaminoglycan (GAG). TGF-β can stimulate sulfated GAG production in nucleus pulposus cells; however, the underlying mechanisms are poorly understood. CHSY1 expression was examined in rat nucleus pulposus treated with TGF-β using real-time PCR and Western blot analysis. Lentiviral knockdown was performed to determine the downstream effectors of TGF-β and to measure the effect of c-Jun and Sp1 on TGF-β mediated CHSY1 promoter activity and CHSY1 expression. TGF-β increased CHSY1 expression and promoter activity in the nucleus pulposus partially through activation of canonical Smad signaling pathway. Knockdown of c-Jun and Sp1 decreased CHSY1 promoter activity, CHSY1 expression and sGAG accumulation induced by TGF-β. Furthermore, we found that TGF-β-induced expression of CHSY1 was mediated through the activation of MAPK signaling. Moreover, we showed that silencing CHSY1 decreased sGAG accumulation in nucleus pulposus cells induced by TGF-β.

Our results suggest that TGF-β induced CHSY1 expression in the nucleus pulposus through the activation of MAPK signaling.

Should endometrial hyperplasia be regarded as a reason for abnormal uterine bleeding in users of the intrauterine contraceptive device?

Histopathological evaluation of the endometrium in cases having an intrauterine device (IUD) removed for abnormal uterine bleeding. Fifty-eight consecutive patients with a complaint of uterine bleeding leading to IUD removal were recruited for this study. Endometrial sampling, via Novak curette, was performed during IUD removal for histopathological evaluation of the endometrium. A total of 58 current IUD users, presenting with uterine bleeding, were retrospectively analyzed. Mean age of the women was 37.1 +/- 2.1 years (range 25-43). Mean duration of IUD use was 4.2 +/- 1.3 years (range 1-10). Out of 58 cases, 21 (36.2%) did not harbor any endometrial pathology. In eight cases (13.8%), hyperplastic endometrial changes were apparent, six of which were simple hyperplasia (four of them were focal). Two cases of complex hyperplasia were detected (one was diffuse with atypia and one was focal without atypia). In this series, there was one case with an endometrial polyp. Mean age of cases with hyperplastic endometrium was found to be statistically high compared to those with normal endometrial histology (42.4 +/- 3.2 vs. 37.6 +/- 2.1, p = 0.04). In cases with IUD use of>5 years, chronic endometritis was more prevalent, compared to those with<5 years of use (chi(2) 5.08, p = 0.02).

IUD use is a risk factor for chronic endometritis. Nevertheless, in 13.8% of cases in this series, as a reason for abnormal uterine bleeding, other than endometritis, endometrial hyperplasia constituted the second most common endometrial pathology among cases over the age of 40, on current IUD use and having complaints of abnormal uterine bleeding. Hence, this finding should prompt the physician to perform endometrial sampling in users of an IUD over the age of 40, presenting with abnormal uterine bleeding.

Are there racial differences in breast cancer treatments and clinical outcomes for women treated at M.D. Anderson Cancer Center?

To determine the influence of race on breast cancer treatment and on recurrence and breast cancer specific death. The study population consisted of 6,054 African-American or white women who were diagnosed with breast cancer and received at least one of the treatments including mastectomy or breast conservative surgery, radiation, adjuvant chemotherapy, neo-adjuvant chemotherapy, and adjuvant endocrine therapy at M.D. Anderson Cancer Center between June 1997 and February 2005. The clinical outcomes were disease-free survival and breast-cancer-specific survival. Logistic regression analysis was performed to investigate if race was associated with the selection of each primary treatment while adjusting for tumor characteristics at diagnosis. Cox proportional hazards model was used to determine the effect of race on recurrence-free survival and breast-cancer-specific survival controlling for tumor characteristics, presence of co-morbidity conditions and use of these treatments. The use of any primary treatment for breast cancer was not significantly different by race after adjusting for tumor characteristics and co-morbidity conditions. Although tumor characteristics at diagnosis explained the major differences in clinical outcomes, race remained an independent prognostic factor for breast-cancer-specific survival (P=0.002), and a marginally significant factor for disease-free survival (P=0.063) in multivariate analyses.

Equal treatment may not lead to equal clinical outcomes given similar tumor characteristics at diagnosis. To reduce racial differences in breast cancer recurrence and survival, it is important to have a better understanding of differences in tumor biology by race and to promote the use of early detection programs among African-American women.

Is hyperglycemia during cardiopulmonary bypass an independent risk factor for mortality in patients undergoing cardiac surgery?

Hyperglycemia is commonly present in the perioperative period in patients undergoing cardiac surgery, even during administration of insulin. A direct relationship between postoperative hyperglycemia and mortality has been established in diabetic patients undergoing cardiac surgery. However, this relationship might be confounded because patients with poor outcome receive more glucogenic drugs postoperatively. We assessed the influence of hyperglycemia (highest glucose level) during cardiopulmonary bypass on perioperative morbidity and mortality in diabetic and nondiabetic patients. We performed a multivariate logistic regression analysis on all diabetic (n = 1579) and nondiabetic (n = 4701) patients undergoing cardiac surgery at the Toronto General Hospital between 1999 and 2001. Boluses of insulin were given during cardiopulmonary bypass when the glucose level exceeded 15 mmol/L, when the serum potassium level exceeded 6.0 mmol/L, or both. Overall mortality was 1.8% (n = 115). A high glucose level during cardiopulmonary bypass was an independent predictor of mortality in both diabetic (odds ratio, 1.20; confidence interval, 1.08-1.32) and nondiabetic (odds ratio, 1.12; confidence interval, 1.06-1.19; per millimole per liter increase in glucose) patients. A high glucose level during cardiopulmonary bypass was also an independent predictor of all major adverse events in both patient groups (odds ratio, 1.06; confidence interval, 1.03-1.09). A high glucose level was not closely related to cardiopulmonary bypass (r = 0.3) or aortic crossclamp times (r = 0.4).

A high peak serum glucose level during cardiopulmonary bypass is an independent risk factor for death and morbidity in diabetic patients and unexpectedly also in nondiabetic patients.

Does myocardial infarction increase ACE2 expression in rat and humans?

Angiotensin converting enzyme (ACE) 2 catalyses the cleavage of angiotensin (Ang) I to Ang 1-9 and of Ang II to Ang 1-7. ACE2 deficiency impairs cardiac contractility and upregulates hypoxia-induced genes, suggesting a link with myocardial ischaemia. We studied the expression of ACE2 after myocardial infarction (MI) in the rat as well as in human failing hearts. Rats were killed at days 1, 3, and 28 after MI, or treated for 4 weeks with the ACE inhibitor ramipril (1 mg/kg). Cardiac gene and protein expression of ACE and ACE2 were assessed by quantitative real-time reverse transcriptase-polymerase chain reaction and immunohistochemistry/activity assays/in vitro autoradiography, respectively. Both ACE (P = 0.022) and ACE2 (P = 0.015) mRNA increased in the border/infarct area compared with the viable area at day 3 after MI. By day 28, increases in ACE (P = 0.005) and ACE2 (P = 0.006) mRNA were also seen in the viable myocardium of MI rats compared with myocardium of control rats. ACE2 protein localized to macrophages, vascular endothelium, smooth muscle, and myocytes. Ramipril attenuated cardiac hypertrophy and inhibited cardiac ACE. In contrast, ramipril had no effect on cardiac ACE2 mRNA, which remained elevated in all areas of the MI rat heart. Immunoreactivity of both ACE and ACE2 increased in failing human hearts.

The increase in ACE2 after MI suggests that it plays an important role in the negative modulation of the renin angiotensin system in the generation and degradation of angiotensin peptides after cardiac injury.

Is rac1 required for cardiomyocyte apoptosis during hyperglycemia?

Hyperglycemia induces reactive oxygen species (ROS) and apoptosis in cardiomyocytes, which contributes to diabetic cardiomyopathy. The present study was to investigate the role of Rac1 in ROS production and cardiomyocyte apoptosis during hyperglycemia. Mice with cardiomyocyte-specific Rac1 knockout (Rac1-ko) were generated. Hyperglycemia was induced in Rac1-ko mice and their wild-type littermates by injection of streptozotocin (STZ). In cultured adult rat cardiomyocytes, apoptosis was induced by high glucose. The results showed a mouse model of STZ-induced diabetes, 7 days of hyperglycemia-upregulated Rac1 and NADPH oxidase activation, elevated ROS production, and induced apoptosis in the heart. These effects of hyperglycemia were significantly decreased in Rac1-ko mice or wild-type mice treated with apocynin. Interestingly, deficiency of Rac1 or apocynin treatment significantly reduced hyperglycemia-induced mitochondrial ROS production in the heart. Deficiency of Rac1 also attenuated myocardial dysfunction after 2 months of STZ injection. In cultured cardiomyocytes, high glucose upregulated Rac1 and NADPH oxidase activity and induced apoptotic cell death, which were blocked by overexpression of a dominant negative mutant of Rac1, knockdown of gp91(phox) or p47(phox), or NADPH oxidase inhibitor. In type 2 diabetic db/db mice, administration of Rac1 inhibitor, NSC23766, significantly inhibited NADPH oxidase activity and apoptosis and slightly improved myocardial function.

Rac1 is pivotal in hyperglycemia-induced apoptosis in cardiomyocytes. The role of Rac1 is mediated through NADPH oxidase activation and associated with mitochondrial ROS generation. Our study suggests that Rac1 may serve as a potential therapeutic target for cardiac complications of diabetes.

Does sphingosine kinase 1 activation enhance epidermal innate immunity through sphingosine-1-phosphate stimulation of cathelicidin production?

The ceramide metabolite, sphingosine-1-phosphate (S1P), regulates multiple cellular functions in keratinocytes (KC). We recently discovered that production of a key innate immune element, cathelicidin antimicrobial peptide (CAMP), is stimulated via a NF-κB-dependent mechanism that is activated by S1P when S1P is generated by sphingosine kinase (SPHK) 1. We investigated whether pharmacological modulation of SPHK1 activity, using a novel synthetic SPHK1 activator, (S)-methyl 2-(hexanamide)-3-(4-hydroxyphenyl) propanoate (MHP), stimulates CAMP expression. MHP-mediated changes in both S1P and CAMP downstream mediators were analyzed in normal cultured human KC by qRT-PCR, Western immunoblot, ELISA, confocal microscopy for immunohistochemistry, HPLC and ESI-LC/MS/MS, and microbial pathogen invasion/colonization in a human epidermal organotypic model. Treatment with MHP directly activated SPHK1 and increased cellular S1P content in normal cultured human KC. Because MHP did not inhibit S1P lyase activity, which hydrolyses S1P, augumented S1P levels could be attributed to increased synthesis rather than blockade of S1P degradation. Next, we found that exogenous MHP significantly stimulated CAMP mRNA and protein production in KC, increases that were significantly suppressed by siRNA directed against SPHK1, but not by a scrambled control siRNA. NF-κB activation, assessed by nuclear translocation of NF-κB, occurred in cells following incubation with MHP. Conversely, pretreatment with a specific inhibitor of SPHK1 decreased MHP-induced nuclear translocation of NF-κB, and significantly attenuated the MHP-mediated increase in CAMP production. Finally, topical MHP significantly suppressed invasion of the virulent Staphylococcus aureus into murine skin explants.

MHP activation of SPHK1, a target enzyme of CAMP production, can stimulate innate immunity.

Is inspiratory work capacity more severely depressed than inspiratory muscle strength in patients with heart failure : Novel applications for inspiratory muscle training?

We hypothesized that the ability to sustain maximal inspiratory pressure (SPImax) over time as a measure of work capacity may be more severely affected than inspiratory muscle strength (PImax) in patients with heart failure (HF). We retrospectively investigated eighty patients with HF, NYHA II/III/ ambulatoryIV and a (mean±SD) LVEF 27±8%, and compared them to 25 healthy subjects (HS). During a maximal inspiratory manoeuvre from residual volume (RV) to total lung capacity, PImax was measured as the maximum mouth pressure at RV, inspiratory contraction time (ICT) as the time from RV to end of inspiration and SPImax as the area under a pressure-time curve using an electronic pressure manometer with designed-purpose software (Trainair(®), Project Electronics Ltd., London, UK). Exercise capacity was assessed with cardiopulmonary exercise testing and the 6-minute walk test (6MWT). Patients achieved a (mean±SD) peak VO2=15.7±3.4ml/kg/min and 6MWT=338±88m. PImax, ICT and SPImax were reduced in HF pts at 75%***, 61%*** and 52%*** of HS, and correlated with NYHA (r=-0.485***), (r=-0.507**), (r=-0.500***), peakVO2 (r=0.501***), (r=0.655***), (r=0.508***) and 6MWT (r=0.477***), (r=0.345**), (r=0.530***), respectively (*p<0.05, **p<0.01, ***p<0.001).

PImax, ICT and SPImax were impaired in HF patients compared to HS and may be important determinants of exercise capacity. SPImax was severely depressed even in patients with relatively preserved PImax and should be considered as an additional target index for inspiratory muscle training. Equations are provided to predict SPImax in relation to age and exercise capacity in HF.

Does computational analysis of a novel mutation in ETFDH gene highlight its long-range effects on the FAD-binding motif?

Multiple acyl-coenzyme A dehydrogenase deficiency (MADD) is an autosomal recessive disease caused by the defects in the mitochondrial electron transfer system and the metabolism of fatty acids. Recently, mutations in electron transfer flavoprotein dehydrogenase (ETFDH) gene, encoding electron transfer flavoprotein:ubiquinone oxidoreductase (ETF:QO) have been reported to be the major causes of riboflavin-responsive MADD. To date, no studies have been performed to explore the functional impact of these mutations or their mechanism of disrupting enzyme activity. High resolution melting (HRM) analysis and sequencing of the entire ETFDH gene revealed a novel mutation (p.Phe128Ser) and the hotspot mutation (p.Ala84Thr) from a patient with MADD. According to the predicted 3D structure of ETF:QO, the two mutations are located within the flavin adenine dinucleotide (FAD) binding domain; however, the two residues do not have direct interactions with the FAD ligand. Using molecular dynamics (MD) simulations and normal mode analysis (NMA), we found that the p.Ala84Thr and p.Phe128Ser mutations are most likely to alter the protein structure near the FAD binding site as well as disrupt the stability of the FAD binding required for the activation of ETF:QO. Intriguingly, NMA revealed that several reported disease-causing mutations in the ETF:QO protein show highly correlated motions with the FAD-binding site.

Based on the present findings, we conclude that the changes made to the amino acids in ETF:QO are likely to influence the FAD-binding stability.

Is mortality from nonulcer bleeding similar to that of ulcer bleeding in high-risk patients with nonvariceal hemorrhage : a prospective database study in Italy?

Nonulcer causes of bleeding are often regarded as minor, ie, associated with a lower risk of mortality. To assess the risk of death from nonulcer causes of upper GI bleeding (UGIB). Secondary analysis of prospectively collected data from 3 national databases. Community and teaching hospitals. Consecutive patients admitted for acute nonvariceal UGIB. Early endoscopy, medical and endoscopic treatment as appropriate. Thirty-day mortality, recurrent bleeding, and need for surgery. A total of 3207 patients (65.8% male), mean (standard deviation) age 68.3 (16.4) years, were analyzed. Overall mortality was 4.45% (143 patients). According to the source of bleeding, mortality was 9.8% for neoplasia, 4.8% for Mallory-Weiss tears, 4.8% for vascular lesions, 4.4% for gastroduodenal erosions, 4.4% for duodenal ulcer, and 3.1% for gastric ulcer. Frequency of death was not different among benign endoscopic diagnoses (overall P = .567). Risk of death was significantly higher in patients with neoplasia compared with benign conditions (odds ratio 2.50; 95% CI, 1.32-4.46; P < .0001). Gastric or duodenal ulcer significantly increased the risk of death, but this was not related to the presence of high-risk stigmata (P = .368). The strongest predictor of mortality for all causes of nonvariceal UGIB was the overall physical status of the patient measured with the American Society of Anesthesiologists score (1-2 vs 3-4, P < .001).

No data on the American Society of Anesthesiologists class score in the Prometeo study.

Does epidural anesthesia increase the incidences of urinary retention and hesitancy in micturition after ambulatory hemorrhoidectomy?

This randomized, prospective study was designed to evaluate the role of various anesthesias in postoperative urinary retention and hesitancy in micturition in patients receiving hemorrhoidectomy on ambulatory basis. In a randomized order, 128 ambulatory patients, ASA physical status I or II, were divided into two groups to receive hemorrhoidectomy under epidural or local anesthesia. In all patients, the intraoperative intravenous fluid given was limited to 200 ml +/- 2 ml/kg/h of Ringer's lactate solution. Patients were requested to void urine voluntarily before discharge. The incidences of postoperative urinary retention and hesitancy in micturition were evaluated by telephone interview 24 hours after surgery. Neither the incidence of urinary retention, nor the incidence of hesitancy in micturition was significantly different between the two groups. Patients with age over 50 had a significantly higher incidence of hesitancy in micturition than younger patients. The incidence of hesitancy in micturition seemed higher in male patients (31.3%) than that in females (15.6%), but the difference was not statistically different (P = 0.0585).

With judicious intraoperative fluid restriction and voluntary voiding before discharge, epidural anesthesia does not increase the incidence of postoperative urinary retention or hesitancy in micturition following ambulatory hemorrhoidectomy.

Does low paraoxonase activity predict coronary events in the Caerphilly Prospective Study?

The hypothesis that paraoxonase (PON1) has a role in preventing atherosclerosis is based on experimental, transgenic, and case-control studies but has not previously been tested prospectively. The Caerphilly Prospective Study is a cohort study of men aged 49 to 65 years observed for coronary heart disease (CHD) events (fatal and nonfatal myocardial infarction) over a mean period of 15 years. Serum PON1 activity toward paraoxon was measured in 1353 participants. PON1 activity was 20% lower in the 163 men who had a coronary event (P=0.039). Men in the highest quintile of PON1 activity had a decreased risk compared with those in the lowest quintile (OR 0.57 [95% CI, 0.33 to 0.96]). The inverse relationship between quintiles of serum PON1 activity and CHD risk was graded, the median change in OR across each quintile being 0.87 (0.77 to 0.98). After adjustment for all other CHD risk factors, including HDL cholesterol, this median value became 0.90 (0.78 to 1.02). PON1 was most predictive of a new CHD event in patients at highest risk by virtue of preexisting CHD (adjusted median OR for each quintile, 0.74 [0.59 to 0.93]; n=313) or the presence of other risk factors. For the highest tertile of CHD risk (n=390) calculated by the Framingham equation, adjusted median OR for each quintile was 0.84 (0.66 to 1.05); n=390.

Low serum PON1 activity toward paraoxon is an independent risk factor for coronary events in men at high risk because of preexisting disease or other CHD risk factors.