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Is association of the large multifunctional proteasome ( LMP2 ) gene with Graves ' disease a result of linkage disequilibrium with the HLA haplotype DRB1*0304-DQB1*02-DQA1*0501?

The large multifunctional proteasome (LMP) molecules are over expressed in thyrocytes, the target cells of Graves' disease, and the LMP genes are found within the MHC class II region. The LMP genes may therefore play a role in susceptibility to Graves' disease. The aim of this this study was to determine whether polymorphisms of the LMP genes, LMP 2 and LMP 7 are in linkage disequilibrium with Graves' disease. Target DNA was amplified using the polymerase chain reaction. The distribution of an Arg-His polymorphism in the LMP 2 gene and a G/T polymorphism in the LMP 7 gene, both of which lead to the presence of an HhaI restriction site, were investigated in a population based case control and family based study in patients with Graves' disease. We obtained DNA from 306 patients with Graves' disease and 364 control subjects for the population based case-control study. In an independent family based study, DNA was obtained from 129 families including both parents, an affected sibling with Graves' disease and an unaffected sibling. All families, patients and control subjects were white caucasians. Frequencies of the alleles and genotypes of the LMP 2 and LMP 7 genes were compared between patients and control subjects using the chi2 test. Transmission of alleles from heterozygous parents to affected and unaffected offspring was assessed using the transmission disequilibrium test (TDT). In the case control study, no difference in allele or genotype frequency was seen between patients and control subjects at the LMP7 locus. At the LMP2 locus the R allele and the RH genotype were increased in subjects with Graves' disease when compared with control subjects (R allele: 36.3% vs. 29.5%, pc = 0. 0164; RH genotype: 56.5% vs. 45%, pc = 0.0102). However, the R allele was in linkage disequilibrium with the associated HLA DRB1*0304-DQB1*02-DQA1*0501 haplotype, delta = 0.102. Within the family based study, no preferential allelic transmission was seen from heterozygote parents to offspring at either loci.

These results show that association of the LMP 2 locus with Graves' disease is due to linkage disequilibrium with the associated HLA haplotype DRB1*0304-DQB1*02-DQA1*0501.

Are inflammatory adverse events associated with disease-free survival after vaccine therapy among patients with melanoma?

Multipeptide vaccines for melanoma may cause inflammatory adverse events (IAE). We hypothesize that IAE are associated with a higher rate of immune response (IR) to vaccination and improved clinical outcomes. Adult patients with resected, high-risk (stage IIB to IV) melanoma were vaccinated with a combination of 12 class I major histocompatibility complex (MHC)-restricted melanoma epitopes, and IAE were recorded. A separate category for hypopigmentation (vitiligo) was also assessed. CD8(+) T cell IR was assessed by direct interferon gamma ELISpot analysis. Overall survival and disease-free survival were analyzed by Cox proportional hazard modeling. Out of 332 patients, 57 developed IAE, the majority of which were dermatologic (minimum Common Terminology Criteria for Adverse Events [CTCAE] grade 3). Most nondermatologic IAE were CTCAE grade 1 and 2. Vitiligo developed in 23 patients (7 %). A total of 174 patients (53 %) developed a CD8(+) response. Presence of IAE was significantly associated with development of IR (70 vs. 49 %, p = 0.005) and with disease-free survival (hazard ratio 0.54, p = 0.043). There were no significant associations relating vitiligo or IR alone with clinical outcomes.

IAE are associated with a higher rate of CD8(+) T cell response after vaccination therapy for high-risk melanoma. Our findings suggest either that antitumor activity induced by class I MHC-restricted peptide vaccines may depend on immunologic effects beyond simple expansion of CD8(+) T cells or that the intrinsic inflammatory response of patients contributes to clinical outcome in melanoma.

Does silencing invariant chains of dendritic cells enhance anti-tumor immunity using small-interfering RNA?

Genetic modification of dendritic cells (DCs) has been used as an effective approach to enhance anti-tumor immunity. RNA interference (RNAi), which can cause the degradation of any RNA in a sequence-specific manner, is a post-transcriptional gene silencing mechanism. In this study, small-interfering RNA (siRNA) specific for the Ii gene was transfected into DCs, and the anti-tumor immunity of Ii-silenced DCs was assessed. The silencing effect of siRNA was evaluated by Western blotting and real-time PCR analyses. In vitro cytotoxic activity of T cells was evaluated using a Cytotox 96(®) non-radioactive cytotoxicity assay kit. The time to tumor onset and the tumor volumes were used as reliable indices to assess the anti-tumor immunity in vivo. To further examine the mechanisms underlying the anti-tumor immunity, flow cytometry analysis was used. The Ii expression of DCs was significantly reduced after Ii siRNA transfection. Significant in vitro anti-tumor ability was exhibited when DCs were co-transfected with Ii siRNA plus endogenous tumor antigen (P < 0.05). Furthermore, tumor growth was greatly inhibited when mice were immunized with DCs transfected with Ii siRNA plus tumor antigen prior to or subsequent to tumor implantation. Flow cytometry analysis in vitro and in vivo indicated that both CD4(+) and CD8(+) T cells were significantly activated in the Ii siRNA group (P < 0.05).

Silencing of the Ii gene of DCs may offer a potential approach to enhance DC-based anti-tumor immunity.

Does ingestion of ethanol just prior to sleep onset impair memory for procedural but not declarative tasks?

The aim of Experiment 1 was to determine if moderate ethanol consumption at bedtime would result in memory loss for recently learned cognitive procedural and declarative tasks. The aim of Experiment 2 was to establish that the memory loss due to alcohol consumption at bedtime was due to the effect of alcohol on sleep states. In Experiment 1, participants were asked to learn a cognitive procedural task and a declarative task in the evening. Then, either the same evening or 2 nights later, they were asked to drink ethanol (0.7g/kg). Sleep was monitored for 3 days and re-testing of the tasks was done on the eighth day after training at the same time of day. In Experiment 2, subjects were asked to learn a cognitive procedural task (Tower of Hanoi) and a motor procedural task (Pursuit Rotor) in the late afternoon. Then one group was asked to drink ethanol (0.9 g/kg) right after task acquisition (5 hours before bed), while the other was asked to drink the same dose of ethanol just prior to bedtime. Re-testing was done 8 days later at the same time of day. Subjects in Experiment 1 were 15 college students between the ages of 19 and 24 that appeared to be in good health and were relatively naive in terms of drinking alcohol. Subjects in Experiment 2 were 13 college students in the same age range. These subjects were considered to be more experienced drinkers than subjects in Experiment 1 but were not judged to be heavy drinkers. In Experiment 1, the alcohol ingestion resulted in memory loss for the cognitive procedural task but not the declarative task. Further, the effect was seen when alcohol ingestion occurred the same day or 2 days after the end of acquisition. In Experiment 2, alcohol ingestion at bedtime impaired memory for the cognitive procedural and motor procedural tasks. By contrast, alcohol ingestion in the afternoon, immediately after the acquisition of these two tasks, did not impair memory. There were clear changes in the nature of rapid eye movement (REM) sleep as a result of evening alcohol ingestion. In Experiment 1, the number of REMs and number of minutes of REM in the first half of the night were reduced. In Experiment 2, the reduction was in REM densities in the first half of the night.

Moderate doses of ethanol can modify the REM sleep architecture by reducing the number of REMs and/or REM densities as well as minutes of REM sleep, particularly in the first half of the night. These modifications result in memory impairment for recently learned cognitive procedural material. Alcohol may also have a subtle effect on Stage 2 sleep as well, since memory for a Stage 2 sensitive motor procedural task was also impaired.

Does spring-assisted cranioplasty alter the growth vectors of adjacent cranial sutures?

Expansile cranial springs are used to treat selected cases of sagittal synostosis. After sagittal suturectomy, springs are placed perpendicular to the line of the synostosis. Normal growth across the coronal suture is approximately perpendicular to the suture line. To allow cranial expansion in the parietal direction, it is hypothesized that the normal growth vectors across the coronal sutures are altered. Ten New Zealand White rabbits underwent suturectomy of the midline posterior frontal suture followed by insertion of an expansile spring. Radiologic markers were placed on either side of the normal coronal sutures (n = 20) perpendicular to the released suture. Serial radiology was performed up to 7 weeks. Growth vectors across the coronal suture were compared with those of 10 control rabbits. Dorsal cranial expansion was predominantly in an anterolateral direction in controls. The normal growth vector across the coronal suture occurred at an angle of -1 degree to the midline. A spring altered this growth vector to 63 degrees. Over 7 weeks, the cranial width increased 5.1 mm in rabbits with springs compared with 0.2 mm in the control group (p < 0.01). The increase in cranial length across the coronal suture was 1.82 mm in controls and 0.93 mm in rabbits with springs (p = 0.019).

Application of a spring across a midline cranial suture alters the growth vector of the normal adjacent sutures. The spring greatly increases cranial width at the expense of some of the normal increase in cranial length over this period. This would be beneficial in correcting the cranial index in sagittal synostosis.

Does propranolol reduce variceal pressure and wall tension in schistosomiasis presinusoidal portal hypertension?

Although prophylaxis with beta-blockers has been shown to decrease variceal pressure and wall tension in cirrhotic patients, this has not been demonstrated in non-cirrhotic portal hypertension caused by Schistosoma mansoni infection. Thirteen patients without history of previous gastrointestinal bleeding were included. All of them had high-risk esophageal varices at endoscopy. An endoscopic gauge and a high-frequency endoscopic ultrasonography miniprobe were used to assess transmural variceal pressure and wall tension before and after achieving beta-blockade with propranolol. Baseline variceal pressure decreased from 13.3 +/- 3.5 to 8.2 +/- 2.0 mmHg (P < 0.0001) and wall tension from 500.2 +/- 279.8 to 274.0 +/- 108.3 mg.mm(-1). The overall effect of propranolol on decreasing variceal pressure and wall tension expressed in percentage change in relation to baseline values was 35.7 +/- 18.4% and 35.9 +/- 26.7%, respectively (P = 0.9993).

Propranolol significantly reduced variceal pressure and wall tension in schistosomiasis.

Does proportion of fibrin and platelets differ in thrombi on ruptured and eroded coronary atherosclerotic plaques in humans?

To determine the proportion of platelets and fibrin in coronary thrombi. Immunohistochemical and morphometric means to examine the coronary arteries of 31 patients who died of acute myocardial infarction. Fresh thrombi were detected in the feeding arteries of infarction areas in 23 cases (74%) and were associated with plaque rupture in 18 (78%) and plaque erosion in 5 (22%). An immunohistochemical study showed that the thrombi consisted of a mixture of fibrin and platelets as well as some other types of blood cells. The fibrin and platelet positive areas in the thrombi associated with plaque rupture accounted for 74 (19)% and 35 (20)% (p < 0.01) and those associated with erosion accounted for 51 (6)% and 70 (21)%, respectively, of the total areas. Areas of positive immunoreactivity for tissue factor and C reactive protein were also significantly greater in ruptured than in eroded plaques.

These results indicate that the proportions of fibrin and of platelets differ in coronary thrombi on ruptured and eroded plaques. Higher proportions of tissue factor and C reactive protein contribute more significantly to thrombus formation on plaque rupture than on plaque erosion.

Analysis of third body particles generated during total knee arthroplasty: is metal debris an issue?

There is a concern amongst knee surgeons that during total knee arthroplasty (TKA) certain particles could be generated which might become left inside the knee, and which could become a possible cause of third body wear. Especially during the sawing process, when an oscillating saw is used through slots in a cutting block, there is a potential cause for metal debris generation. So far however, no data exist to substantiate and quantify the amount of metal debris left in situ after TKA, and it was the purpose of this study to evaluate this. In seven consecutive patients undergoing primary cemented TKA, we measured the amount of debris that was present at the end of the operation immediately before closure. The debris was retrieved using pulsed irrigation with 1 l of normal saline, which was then collected and centrifuged. Obtained pellets were subsequently washed and chemically treated to quantify the different components present in the debris. Overall an average amount of 134.9 mg (range 73.6-198.0 mg) debris was retrieved. The debris contained on average 75.8 mg of bone particles (range 41.2-109.3 mg), 57.2 mg (range 31.2-83.9 mg) of cement particles, and 1.96 mg (range 0-7.2 mg) of metal particles. On average the total amount of debris consisted for 56.5% of bony fragments, 42% cement fragments and 1.5% metal particles.

Even despite precise surgical technique including pressurized irrigation, a significant amount of debris is still present after TKA. The debris consists predominantly of bone and cement particles. The presence of metal debris is limited, and contributes only 1.5% to the total amount.

Are cytokine polymorphisms associated with poor sleep maintenance in adults living with human immunodeficiency virus/acquired immunodeficiency syndrome?

Cytokine activity and polymorphisms have been associated with sleep outcomes in prior animal and human research. The purpose of this study was to determine whether circulating plasma cytokines and cytokine polymorphisms are associated with the poor sleep maintenance commonly experienced by adults living with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Cross-sectional descriptive study. HIV clinics and community sites in the San Francisco Bay area. A convenience sample of 289 adults (193 men, 73 women, and 23 transgender) living with HIV/AIDS. None. A wrist actigraph was worn for 72 h to estimate the percentage of wake after sleep onset (WASO%) and total sleep time (TST), plasma cytokines were analyzed, and genotyping was conducted for 15 candidate genes involved in cytokine signaling: interferon-gamma (IFNG), IFNG receptor 1 (IFNGR1), interleukins (IL1B, IL1R2, IL1R2, IL2, IL4, IL6, IL8, IL10, IL13, IL17A), nuclear factor of kappa light polypeptide gene enhancer in B cells (NFKB1 and NFKB2), and tumor necrosis factor-alpha (TNFA). Controlling for demographic variables such as race and sex, and clinical variables such as CD4+ count and medications, higher WASO% was associated with single nucleotide polymorphisms (SNPs) of IL1R2 rs11674595 and TNFA rs1041981 and less WASO% was associated with IL2 rs2069776. IL1R2 rs11674595 and TNFA rs1041981 were also associated with short sleep duration.

This study strengthens the evidence for an association between inflammation and sleep maintenance problems. In this chronic illness population, cytokine polymorphisms associated with wake after sleep onset provide direction for intervention research aimed at comparing anti-inflammatory mechanisms with hypnotic agents for improving sleep maintenance and total sleep time.

Is hTK preservative solution associated with increased biliary complications among patients receiving DCD liver transplants : a single center experience?

This study compares biliary complication rates associated with use of two different preservative solutions, Histidine-Tryptophan-Ketoglutarate (HTK) and University of Wisconsin (UW), utilized in orthotopic liver transplantation (LT) with donations after cardiac death (DCDs). Between 1997-2010, we retrospectively studied 35 LTs performed utilizing DCD donors, preserved either with HTK (n=17) or UW(n=18). Biliary complications were defined by the presence of anastomotic strictures, non-anastomotic strictures, and/or biliary leak on endoscopic retrograde cholangiopancreatography. HTK and UW cohorts were similar in terms of demographics as well as pre- and post-operative biochemical profile. Donor age was significantly higher among HTK compared to UW recipients (41.5 ± 11.9 vs. 26.2 ± 8.8 years, p<0.001). The incidence of post-LT biliary complications was higher in the HTK group (76% vs. 39% in UW group, p=0.041). Hepatic arterial thrombosis (HAT) was observed among 3 HTK patients (17.7%) and 1 UW patient (5.6%), p=0.33. No patients underwent retransplantation in UW group, five recipients in HTK group underwent retransplantation (29%), p=0.019; 4 due to biliary complications and 1 due to HAT.

This single-center study reveals that the use of HTK preservative among DCD donors is associated with an increased risk of biliary complications. Multicenter retrospective studies are suggested to further verify this observation.

Does phase-Contrast Radiography enable Detection of Early Changes in Articular Cartilage in a Mouse Model of Osteoarthritis?

The aim of this study was to investigate the potential of using phase-contrast radiography to detect early changes in articular cartilage in a mouse model of osteoarthritis. An iodoacetate-induced mouse model of osteoarthritis was used. Mice with iodoacetate-induced osteoarthritis were euthanized at 1 and 2 wks after injection. Specimens were obtained by sectioning mouse tissue into 5-mm-thick slices. Sections were examined using synchrotron x-rays from the 7B2 beamline at the Pohang Light Source. Magnified microradiographic images, obtained by phase-contrast radiography, were used to identify changes in articular cartilage and the subchondral bone. Refraction-enhanced phase-contrast radiography enabled detailed microscopic and macroscopic observation of articular cartilage by a noninvasive technique. Detailed microscopic images revealed partially damaged articular cartilage, containing an irregular trilaminar zone, at 1 wk after injection. At 2 wks after injection, a necrotic cleft was clearly visible, along with collapse of the trilaminar zones. Shrunken chondrocytes and chondrocyte clusters of variable sizes were also observed.

Phase-contrast radiography is a feasible technique for the early detection of osteoarthritis and is capable of detecting microstructural changes in articular cartilage.

Does basal release of nitric oxide augment the Frank-Starling response in the isolated heart?

The Frank-Starling response contributes to the regulation of cardiac output. The major underlying subcellular mechanism is a length-dependent change in myofilament responsiveness to Ca2+. Recent studies indicate that nitric oxide decreases myofilament responsiveness to Ca2+ and modulates myocardial relaxation and left ventricular (LV) diastolic function. We therefore investigated the interaction between nitric oxide and the Frank-Starling response. Isolated ejecting guinea pig hearts (constant afterload and heart rate) were studied before and after interventions. Elevation of filling pressure from 10 to 20 cm H2O increased cardiac output, LV end-diastolic pressure (LVEDP), and peak LV pressure (LVPmax). In the presence of N(G)-monomethyl-L-arginine (L-NMMA, 10 micromol/L; n=10) or free hemoglobin (1 micromol/L; n=8), preload-induced increases in cardiac output were significantly attenuated but baseline cardiac output was unaffected. The effects of L-NMMA were inhibited in the presence of excess L-arginine (100 micromol/L; n=6). These changes were not attributable to alterations in coronary flow. Prostaglandin F2alpha (0.01 micromol/L; n=6), which reduced coronary flow, failed to alter the cardiac output response to preload elevation. The exogenous nitric oxide donor sodium nitroprusside (1 micromol/L; n=6) reduced cardiac output at the lowest preload but not at higher preloads. LVEDP was elevated after L-NMMA and hemoglobin but reduced after sodium nitroprusside.

Basal intracardiac production of nitric oxide significantly augments preload-induced rises in cardiac output in the isolated ejecting guinea pig heart. The mechanism appears to be unrelated to changes in coronary flow and may involve direct effects of nitric oxide on myocardial diastolic and/or systolic function.

Valve Selection in End-Stage Renal Disease: Should It Always Be Biological?

When valve replacement is required in patients with end-stage renal disease (ESRD), it is not clear if mechanical or bioprosthetic valve selection is better. We compared outcomes between ESRD patients who underwent either mechanical or biologic valve replacements at our institution. All patients with ESRD who underwent either mitral or aortic valve replacement from 2002 to 2014 at our institution were reviewed (n = 215; mechanical = 64, biological = 151). A Cox proportional hazards model was used to test the hypothesis that a mechanical valve was correlated with improved long-term survival. Among patients younger than 65 years (n = 123) we also compared survival with the Kaplan-Meier method. Similar unadjusted survival was found for patients who received either a bioprosthetic or mechanical valve (log-rank p = 0.55). Survival is clearly attenuated in this patient population, with only about half the patients younger than 65 years surviving beyond 2 years. In the proportional hazards model, a mechanical valve was not correlated with improved survival even when controlled for other variables, including shock, endocarditis, mitral valve replacement, and patient age (95% confidence interval for hazard ratio of mechanical valve: 0.64 to 1.62).

It appears that there is minimal difference in survival after operation for ESRD patients who undergo bioprosthetic or mechanical valve replacement, even in patients younger than 65 years. The attenuated survival of the ESRD population after valve replacement makes the increased burden of anticoagulation (particularly in hemodialysis patients) unattractive. It is likely that only a small portion of ESRD patients benefit from the increased durability of a mechanical valve.

Does gas6/AXL signaling regulate self-renewal of chronic myelogenous leukemia stem cells by stabilizing β-catenin?

Quiescent leukemia stem cells (LSCs) are important resources of resistance and relapse in chronic myelogenous leukemia (CML). Thus, strategies eradicating CML LSCs are required for cure. In this study, we discovered that AXL tyrosine kinase was selectively overexpressed in primary CML CD34+ cells. However, the role of AXL and its ligand Gas6 secreted by stromal cells in the regulation of self-renewal capacity of LSCs has not been well investigated. The function of CML CD34+ cells was evaluated by flow cytometer, CFC/replating, long-term culture-initiating cells (LTC-IC), CML mouse model driven by human BCR-ABL gene and NOD-scid-IL2Rg-/- (NSI) mice. AXL was selectively overexpressed in primary CML CD34+ cells. AXL knockdown reduced the survival and self-renewal capacity of human CML CD34+ cells. Pharmacological inhibition of AXL reduced the survival and self-renewal capacity of human CML LSCs in vitro and in long-term grafts in NSI mice. Human CML CD34+ cells conscripted bone marrow-derived stromal cells (BMDSCs) and primary mesenchymal stem cells (MSCs) to secret Gas6 to form a paracrine loop that promoted self-renewal of LSCs. Suppression of AXL by shRNA and inhibitor prolonged survival of CML mice and reduced the growth of LSCs in mice. Gas6/AXL ligation stabilizes β-catenin in an AKT-dependent fashion in human CML CD34+ cells.

Our findings improve the understanding of LSC regulation and validate Gas6/AXL as a pair of therapeutic targets to eliminate CML LSCs.

Equity in interviews: do personal characteristics impact on admission interview scores?

Research indicates that some social groups are disadvantaged by medical school selection systems. The stage(s) of a selection process at which this occurs is unknown, but at interview, when applicant and interviewer are face-to-face, there is potential for social bias to occur. We performed a detailed audit of the interview process for a single-entry year to a large UK medical school. Our audit included investigating the personal characteristics of both interviewees and interviewers to find out whether any of these factors, including the degree of social matching between individual pairs of interviewees and interviewers, influenced the interview scores awarded. A total of 320 interviewers interviewed 734 applicants, providing complete data for 2007 interviewer-interviewee interactions. The reliability of the interview process was estimated using generalisability theory at 0.82-0.87. For both interviewers and interviewees, gender, ethnic background, socio-economic group and type of school attended had no influence on the interview scores awarded or achieved. Staff and student interviewer marks did not differ significantly. Although numbers in each group of staff interviewers were too small for formal statistical analysis, there were no obvious differences in marks awarded between different medical specialties or between interviewers with varying amounts of interviewing experience.

Our data provide reassurance that the interview does not seem to be the stage of selection at which some social groups are disadvantaged. These results support the continued involvement of senior medical students in the interview process. Despite the lack of evidence that an interview is useful for predicting future academic or clinical success, most medical schools continue to use interviews as a fundamental component of their selection process. Our study has shown that at least this arguably misplaced reliance upon interviewing is not introducing further social bias into the selection system.

Is protamine-induced cardiotoxicity prevented by anti-TNF-alpha antibodies and heparin?

We investigated the role of tumor necrosis factor alpha (TNF-alpha) in protamine-induced cardiotoxicity and the possibility of preventing or decreasing this effect by anti TNF-alpha antibodies and heparin. Isolated rat hearts were perfused for 60 min with Krebs-Henseleit solution (KH). The control group was perfused with KH alone, the KH > protamine > KH group was treated from the 20th to the 40th minute with protamine, and the KH + anti-TNF > protamine + anti-TNF > KH + anti-TNF group was treated the same as the KH > protamine > KH group but with anti-TNF-alpha antibodies added throughout perfusion. The KH + heparin > protamine + heparin > KH + heparin group was treated the same as the KH > protamine > KH group but with heparin added to KH throughout perfusion. The KH > protamine > KH + heparin was perfused the same as the KH> protamine > KH group but with heparin added to KH for the last 20 min. Left ventricular (LV) function and coronary flow were measured every 10 min. TNF-alpha was measured in the coronary sinus effluent. Left ventricular TNF messenger RNA was determined in the control and KH > protamine > KH groups at baseline and after the 40-min perfusion. Protamine caused a significant decrease of peak systolic pressure and dP/dt (to 25% of baseline). Significant amounts of TNF-alpha in the effluent in the KH > protamine > KH group (102.3 +/- 15.5 pg/min) and TNF messenger RNA expression in left ventricular samples were detected. TNF-alpha was below detectable concentrations in the control, KH + anti-TNF > protamine + anti-TNF > KH + anti-TNF, and KH + heparin > protamine + heparin > KH + heparin groups. TNF-alpha concentrations correlated with depression of LV peak systolic pressure (r = 0.984; P = 0.01) and first derivate of the increase of LV pressure (r = 0.976; P = 0.001). Heparin improved LV recovery and decreased protamine-induced TNF-alpha release (KH > protamine > KH + heparin group).

Anti-TNF-alpha antibodies and heparin prevent protamine-induced TNF-alpha release and depression of LV function. Heparin improves protamine-induced depression of cardiac function.

Does pentoxifylline increase gut ketogenesis following trauma and hemorrhagic shock?

Although pentoxifylline produces various beneficial effects following adverse circulatory conditions, it is not known whether this agent has any effects on gut lipid metabolism after trauma-hemorrhage and resuscitation. The aim of this study, therefore, was to determine whether or not administration of pentoxifylline after trauma-hemorrhagic shock has any salutary effects on gut ketogenesis. A prospective, controlled animal study. A university research laboratory. Fifty-six male Sprague-Dawley rats. Rats underwent a midline laparotomy (i.e., trauma-induced) and were bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the shed blood volume was returned in the form of lactated Ringer's solution. The animals were then resuscitated with four times the volume of maximal bleedout with lactated Ringer's solution over 60 mins. Pentoxifylline (50 mg/kg body weight) or an equivalent volume of normal saline was infused intravenously over 100 mins during and after resuscitation. For in vivo lipid loading, one milliliter of olive oil was given intraduodenally on the completion of resuscitation. Blood samples from portal vein and carotid artery, as well as enterocytes from proximal small intestine, were obtained at 1.5 hrs after fat feeding. Mitochondrial fatty acid beta-oxidation enzyme (i.e., palmitoyl-coenzyme A dehydrogenase) activity, as well as portal and arterial plasma beta-hydroxybutyrate values, were determined. Palmitoyl-coenzyme A dehydrogenase activity in villus tip cells and plasma beta-hydroxybutyrate values in portal vein and carotid artery were significantly reduced after trauma-hemorrhage and resuscitation. Pentoxifylline administration, however, significantly increased mitochondrial fatty acid beta-oxidation enzyme activity and portal plasma beta-hydroxybutyrate concentration without significantly affecting arterial concentrations under such conditions.

Pentoxifylline promotes gut ketogenesis following trauma-hemorrhage and resuscitation.

Is kITENIN associated with tumor progression in human gastric cancer?

KAI1 COOH-terminal interacting tetraspanin (KITENIN) promotes tumor cell migration, invasion and metastasis in colon, bladder, head and neck cancer. The aims of current study were to evaluate whether KITENIN affects tumor cell behavior in human gastric cancer cell line and to document the expression of KITENIN in a well-defined series of gastric tumors, including complete long-term follow-up, with special reference to patient prognosis. To evaluate the impact of KITENIN knockdown on behavior of a human gastric cancer cell line, AGS, migration, invasion and proliferation assays using small-interfering RNA were performed. The expression of activator protein-1 (AP-1) target genes and AP-1 transcriptional activity were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and luciferase reporter assay. The expression of KITENIN and AP-1 target genes by RT-PCR and Western blotting or immunohistochemistry was also investigated in human gastric cancer tissues. The knockdown of KITENIN suppressed tumor cell migration, invasion and proliferation in AGS cells. The mRNA expression of matrix metalloproteinase-1 (MMP-1), MMP-3, cyclooxygenase-2 (COX-2), and CD44 was reduced by knockdown of KITENIN in AGS. AP-1 transcriptional activity was significantly decreased by knockdown of KITENIN in AGS cells. KITENIN expression was significantly increased in human cancer tissues at RNA and protein levels. Expression of MMP-1, MMP-3, COX-2 and CD44 were significantly increased in human gastric cancer tissues. Immunostaining of KITENIN was predominantly identified in the cytoplasm of cancer cells. Expression of KITENIN was significantly associated with tumor size, Lauren classification, depth of invasion, lymph node metastasis, tumor stage and poor survival.

These results indicate that KITENIN plays an important role in human gastric cancer progression by AP-1 activation.

Is a one-week course of triple anti-Helicobacter pylori therapy sufficient to control active duodenal ulcer?

Triple therapy currently forms the cornerstone of the treatment of patients with Helicobacter pylori-positive duodenal ulcer.AIM: To establish whether prolonged antisecretory therapy is necessary in patients with active duodenal ulcer. A total of 77 patients with H. pylori-positive duodenal ulcer were included in a prospective, controlled, double-blind study. All patients received a 7-day treatment with omeprazole 20 mg b.d., clarithromycin 500 mg b.d. and amoxicillin 1000 mg b.d. Patients in the omeprazole group underwent an additional 14-day therapy with omeprazole 20 mg; patients in placebo group received placebo. Endoscopy was performed upon inclusion in the study and after 3 and 8 weeks. Seventy-four patients were eligible for a per protocol analysis after 3 weeks, and 65 after 8 weeks. After 3 weeks, the healing rate was 89% in the omeprazole group and 81% in the placebo group (P=0.51). After 8 weeks, the ulcer healed in 97% of the patients in the total group (95% CI: 92.7-100%). H. pylori was eradicated in 88% of patients in the omeprazole group and in 91% in the placebo group (P=1.0). No statistically significant differences between the groups were found in ulcer-related symptoms or in ulcer healing.

In patients with H. pylori-positive duodenal ulcer, a 7-day triple therapy alone is sufficient to control the disease.

Do reactive oxygen metabolites induce a biphasic contractile response in microvascular lung pericytes?

The changes in microvascular permeability characteristic of postinjury inflammation and sepsis may involve dysfunctional regulatory mechanisms at the capillary level. Pericytes, positioned abluminal to microvascular endothelium may, by their contractility, contribute to this regulation. Reactive oxygen metabolites (ROMs), well-known participants in lung inflammation, may exert an effect on pericytes, leading to changes in permeability and adult respiratory distress syndrome. This study investigates the effect of ROMs and antioxidants in an established in vitro assay of pericyte contractility. Rat lung pericytes were cultured on collagen gel matrices. After exposure to the ROMs, the surface area of the collagen disks was digitally quantified (an integrated measure of cellular contraction) at 10 and 30 minutes. The cells were exposed to hydrogen peroxide and pyrogallol at 10, 100, and 1,000 micromol/L. Antioxidant effects of catalase (100 micromol/L), superoxide dismutase (100 micromol/L), and pretreatment with vitamin E (1 mmol/L) were quantified. Hydrogen peroxide and pyrogallol induced concentration-dependent relaxation at 10 minutes. Conversely, concentration-dependent contraction was seen at 30 minutes. Catalase completely attenuated both responses, whereas superoxide dismutase had no effect. Vitamin E had no effect at 10 minutes but partially attenuated the contraction seen at 30 minutes.

ROMs are capable of producing early relaxation and late contraction in cultured lung pericytes. Whereas catalase attenuates both responses, membrane-bound vitamin E only partially attenuates late contraction. This suggests two separate mechanisms: early physiologic relaxation through signaling pathways affecting actin/myosin tone, and late membrane damage causing contraction. Either pathway may cause dysfunction in pulmonary capillary fluid regulation.

Does dexamethasone reduce reintubation rate due to postextubation stridor in a high-risk paediatric population?

To study the effect of dexamethasone on postextubation stridor (PS) incidence and reintubation rate due to PS in a high-risk paediatric intensive care population. All children aged between 4 wk and 6 y, who were intubated for at least 24 h and extubated between August 1999 and May 2002, were retrospectively included (n=60). Medical records of the included patients were studied; records of patients treated with dexamethasone prior to and following extubation (n=23) were compared with control patients who had not received prophylactic medication (n=37). Nine patients in the control group developed significant postextubation stridor, necessitating nebulized epinephrine or glucocorticosteroids. In six of these children, reintubation as a result of postextubation stridor was indicated. None of the patients treated with dexamethasone developed severe postextubation stridor or required reintubation.

The risk of postextubation stridor is relatively high in the group of children aged between 4 wk and 6 y with intubation exceeding 24 h. We found dexamethasone to be effective in preventing reintubation due to postextubation stridor in this paediatric high-risk group.

Is disruption of gallbladder smooth muscle function an early feature in the development of cholesterol gallstone disease?

BACKGROUND; Decreased gallbladder smooth muscle (GBSM) contractility is a hallmark of cholesterol gallstone disease, but the interrelationship between lithogenicity, biliary stasis, and inflammation are poorly understood. We studied a mouse model of gallstone disease to evaluate the development of GBSM dysfunction relative to changes in bile composition and the onset of sterile cholecystitis. BALB/cJ mice were fed a lithogenic diet for up to 8 weeks, and tension generated by gallbladder muscle strips was measured. Smooth muscle Ca(2+) transients were imaged in intact gallbladder. Lipid composition of bile was altered lithogenically as early as 1 week, with increased hydrophobicity and cholesterol saturation indexes; however, inflammation was not detectable until the fourth week. Agonist-induced contractility was reduced from weeks 2 through 8. GBSM normally exhibits rhythmic synchronized Ca(2+) flashes, and their frequency is increased by carbachol (3 μm). After 1 week, lithogenic diet-fed mice exhibited disrupted Ca(2+) flash activity, manifesting as clustered flashes, asynchronous flashes, or prolonged quiescent periods. These changes could lead to a depletion of intracellular Ca(2+) stores, which are required for agonist-induced contraction, and diminished basal tone of the organ. Responsiveness of Ca(2+) transients to carbachol was reduced in mice on the lithogenic diet, particularly after 4-8 weeks, concomitant with appearance of mucosal inflammatory changes.

These observations demonstrate that GBSM dysfunction is an early event in the progression of cholesterol gallstone disease and that it precedes mucosal inflammation.

Does treatment with hydrogen sulfide attenuate sublesional skeletal deterioration following motor complete spinal cord injury in rats?

Treatment with hydrogen sulfide mitigates spinal cord injury-induced sublesional bone loss, possibly through abating oxidative stress, suppressing MMP activity, and activating Wnt/β-catenin signaling. Spinal cord injury (SCI)-induced sublesional bone loss represents the most severe osteoporosis and is resistant to available treatments to data. The present study was undertaken to explore the therapeutic potential of hydrogen sulfide (H SCI was generated by surgical transaction of the cord at the T3-T4 levels in rats. Treatment with NaHS was initiated through intraperitoneal injection of 0.1 ml/kg/day of 0.28 mol/l NaHS from 12 h following the surgery and over 14 subsequent days. H

Treatment with H

Does neuropsychological test performance predict outcome of cognitive behavior therapy for Chronic Fatigue Syndrome and what is the role of underperformance?

A subgroup of patients with Chronic Fatigue Syndrome (CFS) has cognitive impairments, reflected by deviant neuropsychological test performance. However, abnormal test scores can also be caused by suboptimal effort. We hypothesized that worse neuropsychological test performance and underperformance were related to each other and to a smaller reduction in fatigue, functional impairments, physical limitations and higher dropout rates following cognitive behavior therapy (CBT) for CFS. Data were drawn from a previous trial, in which CFS patients were randomized to two conditions; 1) guided self-instruction and additional CBT (n=84) or 2) waiting period followed by regular CBT for CFS (n=85). Underperformance was assessed using the Amsterdam Short Term Memory Test (<84). To test neuropsychological test performance, the Symbol Digit Modalities Task, a simple reaction time task and a choice reaction time task were used. Interaction effects were determined between underperformance and neuropsychological test performance on therapy outcomes. Underperformance was associated to worse neuropsychological test performance, but there were no significant interaction effects of these two factors by therapy on fatigue severity, functional impairments and physical limitations, but there was a significant main effect of underperformance on functional impairments, physical limitations and dropout rates.

Underperformance or neuropsychological test performance was not related to the change in fatigue, functional impairments, and physical limitations following CBT for CFS. However, underperforming patients did drop out more often. Therapists should pay attention to beliefs and behavioral or environmental factors that might maintain underperformance and increase the risk of dropout.

Underage drinking, brief interventions, and trauma patients: Are they really special?

While the relationship between underage drinking and injury has been well established, few studies have examined whether presenting for an acute injury moderates the efficacy of a brief intervention (BI) on alcohol misuse. Patients (aged 14-20 years) in the emergency department screening positive for risky drinking (Alcohol Use Disorders Identification Test-Consumption score) completed a baseline assessment, were randomized to conditions (a standalone computer-delivered BI [n = 277], a therapist-delivered BI [n = 278], or a control condition [n = 281]), and completed a 3-month follow-up. This secondary analysis of Project U-Connect examined regression models (controlling for baseline values) to examine the main effects of injury and the interaction effects of injury by BI condition on alcohol consumption and consequences. Among 836 youth enrolled in the randomized controlled trial (mean age, 18.6 years; 51.6% were male; 79.4% were white), 303 (36.2%) had a primary complaint of intentional or unintentional injury. At baseline, injured patients were more likely to be male (p<0.001) and have higher alcohol consumption (p<0.01), but were less likely to misuse prescription drugs (p = 0.02) than those presenting for medical reasons. Regression models (controlling for baseline values) demonstrated that injury presentation predicted greater alcohol consumption prior to a BI. The computer BI was more effective at reducing alcohol consequences among those presenting with injury than those presenting for other reasons. Injury did not affect the efficacy of the computer BI on alcohol consumption, and injury did not affect the efficacy of the therapist BI on alcohol outcomes.

A therapist or computer BI reduced alcohol consumption and consequences among risky drinkers regardless of reason for emergency department presentation highlighting the opportunity to reach a broad array of youth. Although the therapist BI was not moderated by injury presentation, the computer BI was particularly effective at reducing alcohol consequences among those presenting with injury at 3-month follow-up.

Is haemoglobin A1c even within non-diabetic level a predictor of cardiovascular disease in a general Japanese population : the Hisayama Study?

There is little information about predictive ability of haemoglobin A1c (HbA1c) for cardiovascular disease (CVD) in Asians. To investigate the discriminatory ability of HbA1c to identify subjects who are at greater risk of developing CVD in a prospective study of a defined community-dwelling Japanese population. A total of 2,851 subjects aged 40-79 years were stratified into five groups (HbA1c levels with ≤ 5.0, 5.1-5.4, 5.5-6.4, and ≥ 6.5% and a group with antidiabetic medication) and followed up prospectively for 7 years (2002-2009). During the follow-up, 119 subjects developed CVD. The multivariable-adjusted risk of CVD was significantly increased in subjects with HbA1c levels of 5.5-6.4 and ≥ 6.5% and diabetic medication compared to HbA1c level with ≤ 5.0% (hazard ratio, 2.26 [95% confidence interval, 1.29-3.95] for the 5.5-6.4%; 4.43 [2.09-9.37] for the ≥ 6.5%; and 5.15 [2.65-10.0] for the antidiabetic medication group). With regard to CVD subtype, the positive associations between HbA1c levels and the risk of coronary heart disease (CHD) and ischaemic stroke were also significant, but no such associations were seen for haemorrhagic stroke. The C statistic for developing CVD was significantly increased by adding HbA1c values to the model including other risk factors (0.789 vs. 0762, p = 0.006), and the net reclassification improvement was 0.105 (p = 0.004).

Our findings suggest that elevated HbA1c levels are an independent risk factor for CVD, especially CHD and ischaemic stroke, and that the addition of HbA1c to the model with traditional risk factors significantly improves the predictive ability of CVD.

Is kinesin involved in regulation of rat pancreatic amylase secretion?

Kinesin has recently been localized to zymogen granules of pancreatic acini and is suggested to participate in exocytosis of exocrine pancreas. We examined the function of kinesin in regulated exocytosis of pancreatic acini in this study. Kinesin function in exocytosis was examined by introducing hexahistidine-tagged recombinant kinesin protein and antikinesin monoclonal antibody into streptolysin-O-permeabilized acini. Intracellular localization of introduced recombinant kinesin was investigated by immunohistochemistry. Interaction between recombinant kinesin and the microtubule network was confirmed by nocodazole pretreatment of acini. Kinesin regulation by secretagogues was investigated by examining their effect on adenosine triphosphatase (ATPase) activity of endogenous kinesin. Recombinant kinesin enhanced calcium-stimulated amylase release from streptolysin-O-permeabilized acini. Introduced recombinant kinesin was localized to both the microtubule network and zymogen granule. Nocodazole pretreatment of acini abolished the enhancing effect of recombinant kinesin on calcium-stimulated amylase release. Antikinesin antibody inhibited amylase release stimulated by the combination of calcium and cyclic adenosine monophosphate (cAMP) but not that stimulated by calcium alone. Secretin and 8-bromo-cAMP increased ATPase activity of endogenous kinesin.

Kinesin plays a stimulatory role in regulated exocytosis of pancreatic acini and is involved in stimulus-secretion coupling through a cAMP-dependent pathway.

Does electro-acupuncture reduce the need for additional anesthetics in experimental studies?

To evaluate the possible beneficial effects of electro-acupuncture in rats subjected to ketamine/xylazine (KX) intra-peritoneal (i.p.) anesthesia. Forty-eight male Wistar rats were distributed in four equal groups. All rats received i.p. injections of ketamine (90 mg/kg) +xylazine (10 mg/kg) anesthesia. Basal values group (control) rats (BV) received no additional treatment. The equivalent of the human right ST36 (Zusanli) and CV-12(Zhongwan) acupoints were chosen for needling and electrical stimulation. AC rats were needled with sterilized disposable stainless steel needles at right ST36 and CV12 acupoints; needles were retained for 30 minutes. EAC10 rats, after needle insertion as described, had electrodes connected to both needles and to an electro stimulator model NKL EL-608; pulsed square waves, 10 Hz, 10 mA, was applied for 30 minutes. EAC100 rats were submitted to EA as described. However, a greater frequency (100 Hz) was used. Thirty-seven rats remained under adequate anesthetic level during the experiment. However, maintenance anesthesia was required by 11 rats. Need for additional anesthesia decreased to 9.1% in EAC100 rats compared to BV (36.3%).

Both the AC and the EAC10/100 prolong the anesthetic effect of the combination Ketamine-xylazine in rats, allowing longer duration of anesthesia with a lower dose of anesthetic, thereby reducing the occurrence of complications.

Does both bias and lack of knowledge influence organizational focus on first case of the day start?

The economic costs of reducing first case delays are often high, because efforts need to be applied to multiple operating rooms (ORs) simultaneously. Nevertheless, delays in starting first cases of the day are a common topic in OR committee meetings. We added three scientific questions to a 24 question online, anonymous survey performed before the implementation of a new OR information system. The 57 respondents cared sufficiently about OR management at the United States teaching hospital to complete all questions. The survey revealed reasons why personnel may focus on the small reductions in nonoperative time achievable by reducing tardiness in first cases of the day. (A) Respondents lacked knowledge about principles in reducing over-utilized OR time to increase OR efficiency, based on their answering the relevant question correctly at a rate no different from guessing at random. Those results differed from prior findings of responses at a rate worse than random, resulting from a bias on the day of surgery of making decisions that increase clinical work per unit time. (B) Most respondents falsely believed that a 10 min delay at the start of the day causes subsequent cases to start at least 10 min late (P < 0.0001 versus random chance). (C) Most respondents did not know that cases often take less time than scheduled (P = 0.008 versus chance). No one who demonstrated knowledge (C) about cases sometimes taking less time than scheduled applied that information to their response to (B) regarding cases starting late (P = 0.0002).

Knowledge of OR efficiency was low among the respondents working in ORs. Nevertheless, the apparent absence of bias shows that education may influence behavior. In contrast, presence of bias on matters of tardiness of start times shows that education may be of no benefit. As the latter results match findings of previous studies of scheduling decisions, interventions to reduce patient and surgeon waiting from start times may depend principally on the application of automation to guide decision-making.

Does leading-process actomyosin coordinate organelle positioning and adhesion receptor dynamics in radially migrating cerebellar granule neurons?

During brain development, neurons migrate from germinal zones to their final positions to assemble neural circuits. A unique saltatory cadence involving cyclical organelle movement (e.g., centrosome motility) and leading-process actomyosin enrichment prior to nucleokinesis organizes neuronal migration. While functional evidence suggests that leading-process actomyosin is essential for centrosome motility, the role of the actin-enriched leading process in globally organizing organelle transport or traction forces remains unexplored. We show that myosin ii motors and F-actin dynamics are required for Golgi apparatus positioning before nucleokinesis in cerebellar granule neurons (CGNs) migrating along glial fibers. Moreover, we show that primary cilia are motile organelles, localized to the leading-process F-actin-rich domain and immobilized by pharmacological inhibition of myosin ii and F-actin dynamics. Finally, leading process adhesion dynamics are dependent on myosin ii and F-actin.

We propose that actomyosin coordinates the overall polarity of migrating CGNs by controlling asymmetric organelle positioning and cell-cell contacts as these cells move along their glial guides.

Does liposomal gene transfer of keratinocyte growth factor improve wound healing by altering growth factor and collagen expression?

Growth factors affect the complex cascade of wound healing; however, interaction between different growth factors during dermal and epidermal regeneration are still not entirely defined. In the present study, we thought to determine the interaction between keratinocyte growth factor (KGF) administered as liposomal cDNA with other dermal and epidermal growth factors and collagen synthesis in an acute wound. Rats received an acute wound and were divided into two groups to receive weekly subcutaneous injections of liposomes plus the Lac-Z gene (0.22 microg, vehicle), or liposomes plus the KGF cDNA (2.2 microg) and Lac-Z gene (0.22 microg). Histological and immunohistochemical techniques were used to determine growth factor, collagen expression, and dermal and epidermal structure. KGF cDNA increased insulin-like growth factor-I (IGF-I), insulin-like growth factor binding protein-3 (IGFBP-3), and fibroblast growth factor (FGF), decreased transforming growth factor-beta (TGF-beta), while it had no effect on platelet-derived growth factor (PDGF) levels in the wound. KGF cDNA significantly increased collagen Type IV at both the wound edge as well as the wound bed, while it had no effect on collagen Type I and III. KGF cDNA increased re-epithelialization, improved dermal regeneration, and increased neovascularization.

Exogenous administered KGF cDNA causes increases in IGF-I, IGF-BP3, FGF, and collagen IV and decreases TGF-beta concentration. KGF gene transfer accelerates wound healing without causing an increase in collagen I or III.

Are levels of drug and antidrug antibodies associated with outcome of interventions after loss of response to infliximab or adalimumab?

There is controversy about whether levels of anti-tumor necrosis factor (TNF) and antidrug antibodies (ADAs) are accurate determinants of loss of response to therapy. We analyzed the association between trough levels of anti-TNF agents or ADAs and outcomes of interventions for patients with loss of response to infliximab or adalimumab. We performed a retrospective study of pediatric and adult patients with inflammatory bowel disease and suspected loss of response to anti-TNF agents treated at medical centers throughout Israel from October 2009 through February 2013. We examined the correlation between outcomes of different interventions and trough levels of drug or ADAs during loss of response. An additional subanalysis was performed including only patients with a definite inflammatory loss of response (clinical worsening associated with increased levels of C-reactive protein or fecal calprotectin, or detection of inflammation by endoscopy, fistula discharge, or imaging studies). Among 247 patients (42 with ulcerative colitis), there were 330 loss-of-response events (188 to infliximab and 142 to adalimumab). Trough levels of adalimumab greater than 4.5 mcg/mL and infliximab greater than 3.8 mcg/mL identified patients who failed to respond to an increase in drug dosage or a switch to another anti-TNF agent with 90% specificity; these were set as adequate trough levels. Adequate trough levels identified patients who responded to expectant management or out-of-class interventions with more than 75% specificity. Levels of antibodies against adalimumab >4 microgram per mL equivalent (mcg/mL-eq) or antibodies against infliximab >9 mcg/mL-eq identified patients who did not respond to an increased drug dosage with 90% specificity. Patients with high titers of ADAs had longer durations of response when anti-TNF agents were switched than when dosage was increased (P = .03; log-rank test), although dosage increases were more effective for patients with no or low titers of ADAs (P = .02). An analysis of definite inflammatory loss-of-response events (n = 244) produced similar results; patients with adequate trough levels had a longer duration of response when they switched to a different class of agent than when anti-TNF was optimized by either a dosage increase or by a switch within the anti-TNF class (P = .002; log-rank test).

The results of this retrospective analysis suggest that trough levels of drug or ADAs may guide therapeutic decisions for more than two-thirds of inflammatory bowel disease patients with either clinically suspected or definite inflammatory loss of response to therapy.

Does pioglitazone improve endothelial and adipose tissue dysfunction in pre-diabetic CAD subjects?

To investigate the effect of pioglitazone on endothelial and adipose tissue dysfunction in newly detected IGT patients with CAD. Participants (n=25) were randomized to treatment with either placebo or pioglitazone (30 mg/day) for 12 weeks. Before and after treatment we evaluated endothelial function (flow-mediated dilation--FMD--of the brachial artery), circulating adipose and inflammatory markers (adiponectin isoforms, TNF-alpha, and high sensitivity-CRP), and insulin sensitivity (euglycemic hyperinsulinemic clamp). No significant changes were observed in subjects (n=12) treated with placebo. By contrast, subjects (n=13) treated with pioglitazone had significant improvement in FMD (10.8±5.3 vs 13.3±3.6%, p<0.01), accompanied by increased high molecular weight adiponectin (HMW-Ad) (1.7±1.2 vs 4.8±3.6 μg/ml, p<0.05) and decreased TNF-alpha (4.3±1.9 vs 3.2±1.2 pg/ml, p<0.05) associated to an increased glucose disposal (4.8±1.9 vs 5.4±2.0 mg kg(-1) min(-1), p<0.05). A multiple regression analysis indicated that increasing of HMW-Ad after pioglitazone predicted increased FMD.

Pioglitazone significantly improves endothelial and adipose tissue dysfunction in pre-diabetic patients with CAD.

Does a philosophical perspective support the need for patient-outcome studies in diagnostic test evaluation?

Most clinical research evaluates diagnostic tests by accuracy measures such as sensitivity and specificity. This is stimulated by the Standards for Reporting of Diagnostic Accuracy initiative, which focuses on accuracy in the widely accepted guidelines. Referring to the clinical consequences of diagnostic tests, many epidemiologists recognize the importance of patient outcome studies in addition to accuracy studies. However, there is a theoretical argument that stipulates the need for patient outcome studies, which has thus far been overlooked. Using a philosophical argument, we show that the definition of disease necessarily involves the concept of function, which in turn is inextricably related to outcome. Consequently, diagnostic tests that establish the presence or absence of disease cannot be evaluated without measuring patient outcome. Patient outcome studies are therefore the definitive means to assess the merits of a diagnostic test.

The need for patient outcome studies is not due to pragmatic reasons, as previous authors argued, but is based on a philosophical argument relating the definition of disease to the concept of function. We propose that authors justify the use of accuracy studies in papers reporting diagnostic test evaluation by describing the association between gold standard test and patient outcome.

The amphetamine sensitization model of schizophrenia: relevance beyond psychotic symptoms?

A sensitized dopamine system may be linked to the genesis of psychotic symptoms in schizophrenia. Following withdrawal from amphetamine exposures, psychotic-like traits have been robustly demonstrated, but the presence of cognitive/mnemonic deficits remains uncertain. Adult male Lewis and Fischer rats, differing in cognitive performance, were exposed intermittently to escalating doses of amphetamine over 5 weeks. This was effective in producing behavioral sensitization to a subsequent amphetamine challenge. Following 27 days of drug withdrawal, the animals were assessed in Pavlovian conditioning, object recognition, and spatial working memory. In addition, prepulse inhibition (PPI), spontaneous motor activity, and anxiety-like behavior were measured. Amphetamine pretreatment induced behavioral sensitization in both rat strains similarly. Working memory was enhanced in Fischer but not Lewis rats following withdrawal. Spontaneous novel object preference was enhanced in sensitized Fischer rats, but was impaired in sensitized Lewis rats, thus effectively reversing the strain difference in non-sensitized controls. In contrast, Pavlovian fear conditioning remained unaffected and so were anxiety-like behavior, open field activity, and PPI.

The face validity of the amphetamine withdrawal model for cognitive deficits was limited to the object recognition memory impairment observed in sensitized Lewis rats. Yet, the possibility that enhancing dopaminergic neurotransmission may facilitate object recognition and spatial working memory performance was demonstrated in sensitized Fischer rats. Identification of the mechanisms underlying such strain-dependent effects would be instrumental in the further specifications of the construct validity, and therefore the limitations and potential of the amphetamine sensitization model of schizophrenia.

Does lentivirus-mediated shRNA interference targeting SLUG inhibit lung cancer growth and metastasis?

Lung cancer is a deadly cancer, whose kills more people worldwide than any other malignancy. SLUG (SNAI2, Snail2) is involved in the epithelial mesenchymal transition in physiological and in pathological contexts and is implicated in the development and progression of lung cancer. We constructed a lentivirus vector with SLUG shRNA (LV-shSLUG). LV-shSLUG and a control lentivirus were infected into the non-small cell lung cancer cell A549 and real-time PCR, Western blot and IHC were applied to assess expression of the SLUG gene. Cell proliferation and migration were detected using MTT and clony formation methods. Real-time PCR, Western Blot and IHC results confirmed down-regulation of SLUG expression by its shRNA by about 80%~90% at both the mRNA and protein levels. Knockdown of SLUG significantly suppressed lung cancer cell proliferation. Furthermore, knockdown of SLUG significantly inhibited lung cancer cell invasion and metastasis. Finally, knockdown of SLUG induced the down-regulation of Bcl-2 and up-regulation of E-cadherin.

These results indicate that SLUG is a newly identified gene associated with lung cancer growth and metastasis. SLUG may serve as a new therapeutic target for the treatment of lung cancer in the future.

Does 2-Methoxyestradiol inhibit hepatocellular carcinoma cell growth by inhibiting Cdc25 and inducing cell cycle arrest and apoptosis?

2-Methoxyestradiol (2-ME) is a physiological metabolite of estrogen, which can inhibit growth of many types of tumor cells, including hepatocellular carcinoma, both in vitro and in vivo. The exact mechanisms of its action are still unclear. We have studied the mechanisms of growth inhibition of several of human and rat hepatoma and normal liver cells by 2-ME. Human (Hep3B, HepG2, PLC/PRF5) and rat (McA-RH7777, JM-1) hepatoma and normal rat (CRL-1439) and human (CRL-11233) liver cell lines were cultured in vitro, in presence of 2-ME, and its IC50s were determined. Cell cycle arrest, Cdc25 phosphatase inhibition and apoptosis induction were studied. Finally, the effect of 2-ME on the growth of JM-1 rat hepatoma cells in rat liver was determined in vivo. The IC50 range for growth inhibition of hepatoma cells was found to be between 0.5 and 3 microM. In contrast, normal rat hepatocytes and liver cell lines were resistant to 2-ME up to 20 microM. JM-1 cells were arrested in the G2/M phase of the cell cycle. Cdc25A and Cdc25B, cell cycle controlling phosphatases, activities were inhibited in vitro and 2-ME was found to likely bind to their catalytic site cysteines. As a consequence, their cellular substrates Cdk1 and Cdk2 were tyrosine phosphorylated. Caspase-3 was cleaved suggesting apoptotic cell death. Moreover, growth of JM-1 tumors, which were transplanted into rat liver, was also inhibited by treatment with 2-ME in vivo.

2-Methoxyestradiol is a selective, potent and relatively non-toxic hepatoma growth inhibitor both in vitro and in vivo. Cell cycle arrest of hepatoma cells was likely mediated by binding and inactivation of the Cdc25 phosphatases and induction of apoptosis.

Does ultrastructure of stomatal development in early-divergent angiosperms reveal contrasting patterning and pre-patterning?

Angiosperm stomata consistently possess a pair of guard cells, but differ between taxa in the patterning and developmental origin of neighbour cells. Developmental studies of phylogenetically pivotal taxa are essential as comparative yardsticks for understanding the evolution of stomatal development. We present a novel ultrastructural study of developing stomata in leaves of Amborella (Amborellales), Nymphaea and Cabomba (Nymphaeales), and Austrobaileya and Schisandra (Austrobaileyales), representing the three earliest-divergent lineages of extant angiosperms (the ANITA-grade). Alternative developmental pathways occur in early-divergent angiosperms, resulting partly from differences in pre-patterning and partly from the presence or absence of highly polarized (asymmetric) mitoses in the stomatal cell lineage. Amplifying divisions are absent from ANITA-grade taxa, indicating that ostensible similarities with the stomatal patterning of Arabidopsis are superficial. In Amborella, 'squared' pre-patterning occurs in intercostal regions, with groups of four protodermal cells typically arranged in a rectangle; most guard-mother cells are formed by asymmetric division of a precursor cell (the mesoperigenous condition) and are typically triangular or trapezoidal. In contrast, water-lily stomata are always perigenous (lacking asymmetric divisions). Austrobaileya has occasional 'giant' stomata.

Similar mature stomatal phenotypes can result from contrasting morphogenetic factors, although the results suggest that paracytic stomata are invariably the product of at least one asymmetric division. Loss of asymmetric divisions in stomatal development could be a significant factor in land plant evolution, with implications for the diversity of key structural and physiological pathways.

Does the Finnish lapphund retinal atrophy locus map to the centromeric region of CFA9?

Dogs have the second largest number of genetic diseases, after humans. Among the diseases present in dogs, progressive retinal atrophy has been reported in more than a hundred breeds. In some of them, the mutation has been identified and genetic tests have allowed the identification of carriers, thus enabling a drastic reduction in the incidence of the disease. The Finnish lapphund is a dog breed presenting late-onset progressive retinal atrophy for which the disease locus remains unknown. In this study we mapped the progressive retinal atrophy locus in the Finnish lapphund using a DNA pooling approach, assuming that all affected dogs within the breed share the same identical-by descent-mutation as the cause of the disease (genetic homogeneity). Autosomal recessive inheritance was also assumed, after ruling out, from pedigree analysis, dominant and X-linked inheritance. DNA from 12 Finnish lapphund cases was mixed in one pool, and DNA from 12 first-degree relatives of these cases was mixed to serve as the control pool. The 2 pools were tested with 133 microsatellite markers, 3 of which showed a shift towards homozygosity in the cases. Individual genotyping with these 3 markers confirmed homozygosity for the GALK1 microsatellite only (chromosome 9). Further individual genotyping with additional samples (4 cases and 59 controls) confirmed the association between this marker and the disease locus (p < 0.001). Closely related to this breed are the Swedish lapphund and the Lapponian herder for which a small number of retinal atrophy cases have been reported. Swedish lapphund cases, but not Lapponian herder cases, had the same GALK1 microsatellite genotype as Finnish lapphund cases.

The locus for progressive rod-cone degeneration is known to be close to the GALK1 locus, on the telomeric region of chromosome 9, where the retinal atrophy locus of the Finnish lapphund has been mapped. This suggests that the disease in this breed, as well as in the Swedish lapphund, may correspond to progressive rod-cone degeneration. This would increase the number of known dog breeds having this particular form of progressive retinal atrophy.

STMN-1 gene: a predictor of survival in stage iia esophageal squamous cell carcinoma after Ivor-Lewis esophagectomy?

Prognosis of esophageal squamous cell carcinoma (ESCC) is stage-specific; however, some patients with the same stage have different survival outcomes. Clinically, it is significant to explore the biological marker to predict patient's outcome. We investigated the association between the stathmin1 gene (STMN-1) expression and the prognosis of patients who underwent Ivor-Lewis esophagectomy. A total of 162 patients who suffered from midthoracic stage IIA ESCC and completely resected with Ivor-Lewis esophagectomy were studied for STMN-1 expression by qRT-PCR in fresh-frozen tissue and validated by immunohistochemistry in matched formalin fixed-paraffin embedded tissue samples. STMN-1 level was evaluated as a prognostic factor in ESCC. SPSS 21.0 software was used to analyze the relationship between STMN-1 expression and clinicopathological characteristics and survival probability. The overall 3- and 5-year survival was 72.20 and 42.00 % respectively. Ninety-four patients (58.02 %) experienced disease recurrence with a disease-free interval of 21.50 ± 1.20 months. qRT-PCR result showed that STMN-1 mRNA level in patients who were alive at the end of follow-up was lower compared with patients who died during the follow-up period (p<0.05). Immunohistochemical results showed that 94 patients had STMN-1 protein overexpression (58.02 %), patient with STMN-1 overexpression had worse survival compared with patients who had low STMN-1 expression (p = 0.00). Cox regression analysis revealed that STMN-1 protein expression and T classification are independent prognostic factors.

Even localized ESCC are potential to relapse with poor prognosis. This study demonstrates that STMN-1 level is an independent prognostic factor after Ivor-Lewis esophagectomy. In addition, assessment of STMN-1 level could improve stratification of stage IIA ESCC patients.

Are aortic root dynamics asymmetric?

The presence of conformational changes in the aortic root during the cardiac cycle is well known, but precise information on time-related changes at each level of the root is lacking. High-resolution, 3D sonomicrometry (200 Hz) was applied in an acute sheep model. Twelve crystals were implanted in eight sheep at each base (n = 3), commissure (n = 3), sinotubular junction (n = 3) and ascending aorta (n = 3). Under stable hemodynamic conditions, geometric changes of the perimeter of each sinus of Valsalva, sinus height, and twist and root tilt angles were time-related to left ventricluar (LV) and aortic pressures. Expansion of the perimeter of the three sinuses of Valsalva was homogeneous, but in significantly different proportions (p < 0.001): the right sinus expanded (+32.4 +/- 2.4%) more than the left (+29.3 +/- 3.2%), and more than the non-coronary (NC) sinus (+25.8 +/- 1.7%). A similar pattern was found for aortic root height: right greater than left, and left greater than NC sinus (p < 0.001). This asymmetry resulted in changes of the root's twist and tilt angles. Although the twist deformation was consistent for each sheep, no general pattern was found. The aortic root tilt angle (between the basal plane and the commissural plane) was 16.3 +/- 1.5 degrees at end-diastole (angle oriented posteriorly and to the left). During systole, it was reduced by 6.6 +/- 0.5 degrees, aligning the LV outflow tract with the ascending aorta. This tilt angle returned to its original value after valve closure.

Aortic root expansion is asymmetric, generating precise changes in its tilt angle. During systole, tilt angle reduction resulted in a straight cylinder that probably facilitates ejection; during diastole, the tilt angle increased, probably reducing leaflet stress. These findings should impact upon surgical procedures and the design of new prostheses.

Is chronic fatigue syndrome associated with chronic enterovirus infection of the stomach?

The aetiology for chronic fatigue syndrome (CFS) remains elusive although enteroviruses have been implicated as one of the causes by a number of studies. Since most CFS patients have persistent or intermittent gastrointestinal (GI) symptoms, the presence of viral capsid protein 1 (VP1), enterovirus (EV) RNA and culturable virus in the stomach biopsy specimens of patients with CFS was evaluated. 165 consecutive patients with CFS underwent upper GI endoscopies and antrum biopsies. Immunoperoxidase staining was performed using EV-specific monoclonal antibody (mAb) or a control mAb specific for cytomegalovirus (CMV). RT-PCR ELISA was performed on RNA extracted from paraffin sections or samples preserved in RNA later. Biopsies from normal stomach and other gastric diseases served as controls. 75 samples were cultured for EV. 135/165 (82%) biopsies stained positive for VP1 within parietal cells, whereas 7/34 (20%) of the controls stained positive (p< or =0.001). CMV mAb failed to stain any of the biopsy specimens. Biopsies taken from six patients at the onset of the CFS/abdominal symptoms, and 2-8 years later showed positive staining in the paired specimens. EV RNA was detected in 9/24 (37%) paraffin-embedded biopsy samples; 1/21 controls had detectable EV RNA (p<0.01); 1/3 patients had detectable EV RNA from two samples taken 4 years apart; 5 patient samples showed transient growth of non-cytopathic enteroviruses.

Enterovirus VP1, RNA and non-cytopathic viruses were detected in the stomach biopsy specimens of CFS patients with chronic abdominal complaints. A significant subset of CFS patients may have a chronic, disseminated, non-cytolytic form of enteroviral infection, which could be diagnosed by stomach biopsy.

Is the prevalence of nonalcoholic steatohepatitis greater in morbidly obese men compared to women?

Nonalcoholic steatohepatitis (NASH) is a form of liver injury that is common in morbidly obese subjects. It has been shown that gender differences exist in the spectrum of nonalcoholic fatty liver disease (NAFLD). The focus of this study was to further characterize these gender differences based on ATP III criteria used to diagnose the metabolic syndrome (MS). We retrospectively assessed NAFLD 58 men and 307 women who underwent gastric bypass, for the presence of NASH, MS, and positive predictors of NASH. There was no statistical difference in age, gender, or the presence of diabetes. The prevalence of NASH in men and women was 60.3% and 30.9%, respectively (P<0.001). Multivariate logistic analysis showed an association of male gender with NASH (2.7; 95% CI, 1.3-5.6, P=0.006) as well as age, AST, and diabetes. MS was diagnosed in 91.4% and 76.2% of men and women (P=0.008), and men tended to have more criteria for MS compared to women. The only positive predictor of MS that was statistically significant between genders was high triglycerides (P=0.003). Controlling for BMI and excess body weight produced similar results.

Gender differences do exist within NAFLD and MS, that may be associated with free fatty acid flow to the liver.

Does extending ventilator circuit change interval beyond 2 days reduce the likelihood of ventilator-associated pneumonia?

To determine the risk of acquiring ventilator-associated pneumonia (VAP) and the impact on costs when extending ventilator circuit change intervals beyond 2 days to 7 and 30 days. Prospective 4-year review of mechanically ventilated patients. The respiratory and medical ICUs of an 800-bed tertiary teaching Veterans Affairs hospital. All adult patients receiving mechanical ventilation from January 1991 through December 1994. Ventilator circuits with active heated water humidifiers were changed at 2-day intervals during a 2-year control period, followed by 7-day and 30-day intervals (for 1 year each). Heated wire circuits were adopted with the 30-day interval. The rate of VAP per 1,000 ventilator days was calculated for each circuit change interval group. Survival analysis was used to model VAP with ventilator circuit change to determine risk. During the study period, 637 patients received mechanical ventilation. During the 2 years with 2-day change intervals, the VAP per 1,000 ventilator days was 11.88 (n=343), compared with 3.34 (n=137) and 6.28 (n=157) for 7-day and 30-day change intervals, respectively. The risk of acquiring a VAP for those with a circuit change every 2 days was significantly greater (relative risk, 3.1; p=0.0004; 95% confidence interval, 1.662, 5.812) than those with the 7- and 30-day circuit changes. Extending circuit change intervals reduced supply and labor costs averaging $4,231/yr for each ventilator in use.

Circuit change intervals of 7 and 30 days have lower risks for VAP than the 2-day intervals, yielding substantial reductions in morbidity as well labor and supply costs.

Does medial prefrontal cortex infusion of alpha-flupenthixol attenuate systemic d-amphetamine-induced disruption of conditional discrimination performance in rats?

It has been argued that tasks that necessitate the use of context in the service of goal-directed behaviour are disrupted in both schizophrenic patients and in animal analogues by dopamine (DA) manipulation with the prefrontal cortex being implicated. To determine the effects on conditional discrimination performance of direct infusion of the DA D(1)/D(2) receptor antagonist alpha-flupenthixol into the medial prefrontal cortex (mPFC) and of its reversal potential on d-amphetamine-induced task disruption. Conditional discrimination performance in which rats learn to respond on an appropriate lever, conditional upon specific auditory stimuli, was acquired and later tested under the above drug treatment protocol in extinction. Conditional discrimination performance was unaffected by bilateral intra-mPFC alpha-flupenthixol at doses of 12, 24 or 36 microg/microl. A dose of D-amphetamine (1.5 mg/kg) shown previously to disrupt conditional discrimination performance was attenuated by direct PFC infusion of alpha-flupenthixol at doses of 24 and 36 but not 12 microg/microl per site.

These results show that conditional discrimination performance is at least in part mediated by prefrontal DA as local PFC DA antagonism attenuates task performance disruption by the indirect DA agonist d-amphetamine further implicating the role of dysfunctional forebrain DA in cognitive deficits evident in schizophrenia.

An observational study of upper gastrointestinal bleeding in intensive care units: is Helicobacter pylori the culprit?

Upper gastrointestinal bleeding (UGIB) related to stress ulcers was formerly a fearsome complication of intensive care. The incidence of this event has decreased over the years. However, the morbidity, mortality, and causes of UGIB, particularly the etiologic role of Helicobacter pylori infection, are still controversial. Therefore, we prospectively assessed the incidence of UGIB in the intensive care unit (ICU) and evaluated the role of H. pylori infection. A prospective observational study followed by a case-control study. Seven ICUs in the Paris area, five of them located in teaching hospitals. All patients admitted consecutively to seven ICUs during a 1-year period were monitored for signs of clinically relevant UGIB. None. Only cases of endoscopically confirmed UGIB were analyzed. Patients whose hemorrhage originated from the stomach and/or duodenum were tested for H. pylori infection, by means of serology, histologic examination, and stool antigen detection. The possible association between H. pylori and UGIB was examined in a case-control study. Twenty-nine of the 4,341 patients admitted to the seven ICUs during the study period had clinically relevant, endoscopically confirmed UGIB (incidence, 0.67%; 95% confidence interval, 0.56%-0.77%). Ulcers were most frequently observed endoscopically. Patients who bled had a higher Simplified Acute Physiology Score (SAPS II) at admission (mean +/- sd, 47 +/- 14 vs. 36 +/- 28; p<.001). Despite a higher in-ICU mortality rate among patients who bled (73% vs. 16%; p<.001), death was never due to bleeding. H. pylori infection was more frequent in patients who bled than in matched controls (36% vs. 16%; p = .04).

Clinically relevant, endoscopically confirmed UGIB is a rare event in the ICU setting and tends to occur in severely ill patients. H. pylori infection is more frequent in patients with gastroduodenal hemorrhage than in nonbleeding patients.

Is heavy episodic drinking a trait-state : a cautionary note?

Heavy episodic (binge) drinking is common in and problematic for undergraduates. Researchers often assume that an individual's heavy episodic drinking is stable and trait-like. However, this fails to consider fluctuating, state-like variation in heavy episodic drinking. This study proposes and tests a novel conceptualization of heavy episodic drinking as a trait-state wherein the contribution of both trait-like stability and state-like fluctuations are quantified. It was hypothesized that heavy episodic drinking is a trait-state such that individuals have trait-like tendencies to engage in heavy episodic drinking, and state-like differences in the expression of this tendency over time. A sample of 114 first-year undergraduates from a Canadian university completed self-report measures of heavy episodic drinking at 3 time points across 130 days. Hypotheses were tested with repeated-measures analysis of variance (ANOVA), test-retest correlations, and generalizability theory analyses. A substantial proportion of the variance in heavy episodic drinking is attributable to trait-like stability, with a smaller proportion attributable to state-like fluctuations.

The heavy episodic drinker seems characterized by a stable, trait-like tendency to drink in a risky manner, and this trait-like tendency seems to fluctuate in degree of expression over time. Findings complement research suggesting that people have trait-like predispositions that increase their risk for heavy episodic drinking. However, despite this stable tendency to drink heavily, the frequency of heavy episodic drinking appears to be at least partly sporadic or situation dependent. These findings serve as a caution to alcohol researchers and clinicians who often assume that a single assessment of heavy episodic drinking captures a person's usual drinking behavior.

Do silent polymorphisms in the RYR1 gene modify the phenotype of the p.4898 I > T pathogenic mutation in central core disease : a case report?

Central core disease is a congenital myopathy, characterized by presence of central core-like areas in muscle fibers. Patients have mild or moderate weakness, hypotonia and motor developmental delay. The disease is caused by mutations in the human ryanodine receptor gene (RYR1), which encodes a calcium-release channel. Since the RYR1 gene is huge, containing 106 exons, mutation screening has been limited to three 'hot spots', with particular attention to the C-terminal region. Recent next-generation sequencing methods are now identifying multiple numbers of variants in patients, in which interpretation and phenotype prevision is difficult. In a Brazilian Caucasian family, clinical, histopathological and molecular analysis identified a new case of central core disease in a 48-year female. Sanger sequencing of the C-terminal region of the RYR1 gene identified two different missense mutations: c.14256 A > C polymorphism in exon 98 and c.14693 T > C in exon 102, which have already been described as pathogenic. Trans-position of the 2 mutations was confirmed because patient's daughter, mother and sister carried only the exon 98's mutation, a synonymous variant that was subsequently found in the frequency of 013-0,05 of alleles. Further next generation sequencing study of the whole RYR1 gene in the patient revealed the presence of additional 5 common silent polymorphisms in homozygosis and 8 polymorphisms in heterozygosis.

Considering that patient's relatives showed no pathologic phenotype, and the phenotype presented by the patient is within the range observed in other central core disease patients with the same mutation, it was concluded that the c.14256 A > C polymorphism alone is not responsible for disease, and the associated additional silent polymorphisms are not acting as modifiers of the primary pathogenic mutation in the affected patient. The case described above illustrates the present reality where new methods for wide genome screening are becoming more accessible and able to identify a great variety of mutations and polymorphisms of unknown function in patients and their families.

Extracapsular axillary node extension in patients receiving adjuvant systemic therapy: an indication for radiotherapy?

This is a retrospective review into the patterns of failure of 82 patients with Stage II or III breast cancer who had extracapsular extension (ECE) of axillary nodal metastases and who received systemic chemotherapy or hormonal therapy without loco-regional radiation. The clinical records of patients with axillary node positive (T1-T3, N1, 2) Stage II or III breast cancer seen at the London Regional Cancer Centre between 1980-1989 were reviewed. Patients were identified who underwent segmental mastectomy with axillary node dissection or modified radical mastectomy and received adjuvant chemotherapy or tamoxifen but did not undergo loco-regional radiation. Eighty-two patients within this group had pathologic evidence of extracapsular axillary node extension (ECE). For 45 of these patients the extension was extensive, and for the remaining 37 it was microscopic. This ECE-positive group was compared to a subgroup of 172 patients who did not have pathologic evidence of extracapsular axillary node extension but had metastatic carcinoma confined within the nodal capsule. Median age of the 82 ECE-positive patients was 56 years. Twenty-five patients had had a segmental mastectomy, the remainder a modified radical mastectomy. Median actuarial survival was 60 months, with a median disease-free and loco-regional failure-free survival of 38 months. Seventy-eight percent of these patients developed a recurrence, which was loco-regional in 60% (21% local, 21% regional, 2% local and regional, and 16% loco-regional and metastatic). There was a 36% recurrence rate in intact breast, 14% the chest wall following modified radical mastectomy, 7% relapsed in the axilla, 12% in supraclavicular nodes, and 1% in the internal mammary nodes. A comparison of the 82 ECE-positive patients with a group of 172 ECE-negative patients determined that there was a statistically significant difference between the two groups in terms of survival (overall and disease-free) and loco-regional recurrence. Univariate analysis of the entire 254 node-positive patient group revealed extracapsular nodal extension (ECE) to be a prognostically significant factor for actuarial and disease-free survival as well as for loco-regional failure, but ECE did not remain an independently prognostic factor after multivariate analysis. Segmental mastectomy, positive resection margins, and ER negative status increased the risk of loco-regional recurrence within the ECE-positive group.

Extracapsular axillary node extension is a prognostically significant factor for actuarial survival, disease-free survival, and loco-regional failure but not independent of other adverse prognostic factors. It is a marker for increased loco-regional recurrence associated with doubling of breast, chest wall, and supraclavicular recurrence rates. The risk of axillary relapse in patients who have had an adequate level I and II axillary dissection but demonstrate extracapsular extension is low (7%). We recommend breast/chest wall and supraclavicular radiation for all patients with pathologic evidence of such extranodal extension who have had a level I and II axillary dissection regardless of the number of positive axillary nodes. Axillary irradiation should be considered for patients who have had only an axillary sampling or level I axillary dissection.

Is placental size associated with mental health in children and adolescents?

The role of the placenta in fetal programming has been recognized as a highly significant, yet often neglected area of study. We investigated placental size in relation to psychopathology, in particular attention deficit hyperactivity disorder (ADHD) symptoms, in children at 8 years of age, and later as adolescents at 16 years. Prospective data were obtained from The Northern Finland Birth Cohort (NFBC) 1986. Placental weight, surface area and birth weight were measured according to standard procedures, within 30 minutes after birth. ADHD symptoms, probable psychiatric disturbance, antisocial disorder and neurotic disorder were assessed at 8 years (n = 8101), and ADHD symptoms were assessed again at 16 years (n = 6607), by teachers and parents respectively. We used logistic regression analyses to investigate the association between placental size and mental health outcomes, and controlled for gestational age, birth weight, socio-demographic factors and medical factors, during gestation. There were significant positive associations between placental size (weight, surface area and placental-to-birth-weight ratio) and mental health problems in boys at 8 and 16 years of age. Increased placental weight was linked with overall probable psychiatric disturbance (at 8 y, OR= 1.14 [95% CI= 1.04-1.25]), antisocial behavior (at 8 y, OR = 1.14 [95% CI= 1.03-1.27]) and ADHD symptoms (inattention-hyperactivity at 16 y, OR= 1.19 [95% CI = 1.02-1.38]). No significant associations were detected among girls.

Compensatory placental growth may occur in response to prenatal insults. Such overgrowth may affect fetal development, including brain development, and ultimately contribute to psychopathology.

Does [ SHENG MAI ZHUSHEYE improve the viability and movement parameters of human sperm in vitro ]?

To observe the effect of SHENG MAI ZHUSHEYE on the movement parameters and viability of human sperm in vitro. We collected sperm samples from 33 normal fertile men, divided each into two, and cultured them in vitro with SHENG MAI ZHUSHEYE + Hams-F10 and Hams-F10 alone, respectively. Then we measured the straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP) and the amplitude of lateral head displacement (ALH) of the sperm by computer-aided semen analysis at 0.5, 1, 2, 4, 8 and 12 h. And the sperm viability was detected. VCL was significantly higher at 8 h (P < 0.05) and very significantly higher at 12 h (P < 0.01) in the SHENG MAI ZHUSHEYE + Hams-F10 group than in the Hams-F10 group. VSL, VAP and ALH were significantly increased in the former group at 4, 8 and 12 h as compared with the latter (P < 0.05). The sperm viability was significantly decreased in the Hams-F10 group at 12 h (P < 0.05).

SHENG MAI ZHUSHEYE can improve sperm movement parameters and increase sperm viability in vitro.

Does chronic exercise preserve renal structure and hemodynamics in spontaneously hypertensive rats?

Exercise training (ExT) is a recommended adjunct to many pharmaceutical antihypertensive therapies. The effects of chronic ExT on the development of hypertension-induced renal injury remain unknown. We examined whether ExT would preserve renal hemodynamics and structure in the spontaneously hypertensive rat (SHR), and whether these effects were mediated by improved redox status and decreased inflammation. Normotensive WKY rats and SHR underwent moderate-intensity ExT for 16 weeks. One group of SHR animals was treated with hydralazine to investigate the pressure-dependent/independent effects of ExT. Acute renal clearance experiments were performed prior to sacrifice. Tissue free radical production rates were measured by electron paramagnetic resonance; gene and protein expression were measured by real time RT-PCR and Western blot or immunofluorescence, respectively. Plasma angiotensin II levels and kidney antioxidants were assessed. Training efficacy was assessed by citrate synthase activity assay in hind-limb muscle. ExT delayed hypertension, prevented oxidative stress and inflammation, preserved antioxidant status, prevented an increase in circulating AngII levels, and preserved renal hemodynamics and structure in SHR. In addition, exercise-induced effects, at least, in part, were found to be pressure-independent. This study is the first to provide mechanistic evidence for the renoprotective benefits of ExT in a model of hypertension. Our results demonstrate that initiation of ExT in susceptible patients can delay the development of hypertension and provide renoprotection at the functional and ultrastructural level.

Chronic ExT preserves renal hemodynamics and structure in SHR; these effects are partially mediated by improved redox status and decreased inflammation.

Does preservative-free versus preserved latanoprost eye drop in patients with open-angle glaucoma or ocular hypertension?

This study compared the efficacy, safety, and pharmacokinetics of a preservative-free latanoprost formulation to an established, benzalkonium chloride (BAK) containing formulation for the treatment of open-angle glaucoma or ocular hypertension. This was a phase II, randomized, cross-over, investigator-masked, multi-center, pilot study (NCT01494753). A total of 30 untreated adult patients (aged ≥18 years) with primary open angle glaucoma, pseudo-exfoliative glaucoma, pigmentary glaucoma, or ocular hypertension received either preservative-free or preserved latanoprost once daily in both eyes for 6 weeks, before crossing over to receive the other treatment. Efficacy (intraocular pressure [IOP] at 8 am, midday, 4 pm and 8 pm, and global efficacy assessment by investigator), safety (adverse events, ocular symptoms and global tolerance, slit lamp examination, funduscopy, visual field examination, visual acuity, and heart rate), and pharmacokinetics were assessed at Days 0, 42, and 84. Both treatments resulted in a reduction in IOP that was similar for the preservative-free and the preserved formulation at all time points. Similarly, the overall diurnal reduction was similar in both groups (6.3 mmHg [27.9% reduction] and 6.4 mmHg [28.1% reduction] for preserved and preservative-free latanoprost, respectively). There were no differences in global efficacy assessment or in the safety and tolerance of each treatment. Systemic concentrations of latanoprost were very low; AUC0-30 and Cmax were lower and tmax was longer for preservative-free latanoprost.

Preservative-free latanoprost showed similar efficacy at all time points compared to BAK preservative containing formulation, with no difference in tolerance, allowing progression to phase III clinical development.

Are cerebrospinal fluid antibodies to tubulin elevated in the patients with multiple sclerosis?

The aim of this study was to compare the levels of anti-tubulin antibodies (anti-TU) in cerebrospinal fluid (CSF) and serum using bovine tubulin as the antigen in one enzyme-linked immunosorbent assay (ELISA) method (anti-TUb antibodies) and a synthetic neuron-specific octapeptide of tubulin in a second ELISA method (anti-TUs antibodies). Paired CSF and serum samples were obtained from 34 multiple sclerosis (MS) patients, 13 patients with various other neurological diseases (control diseases) and 17 normal control patients (CN). CSF levels of anti-TUs and anti-TUb antibodies were significantly lower in the CN group when compared to those in the MS group. On the contrary, serum levels of anti-TU antibodies did not differ among groups. The intrathecal synthesis of anti-TUs antibodies in comparison with anti-TUb was significantly increased in all groups. Significant correlations between anti-TUb and anti-TUs antibodies were observed in the CSF of all three groups. However, with regard to serum, a similar relationship was only found in the MS group.

The estimation of anti-TU in CSF can contribute to the overall assessment of axonal damage; on the contrary serum anti-tubulin antibodies were not useful for differential purposes in MS. The antibodies to the neuron-specific portion of tubulin seemed to be synthesised predominantly intrathecally.

Does sulfhydration of p66Shc at cysteine59 mediate the antioxidant effect of hydrogen sulfide?

Mitochondrion is considered as the major source of intracellular reactive oxygen species (ROS). H2S has been reported to be an antioxidant, but its mechanism remains largely elusive. P66Shc is an upstream activator of mitochondrial redox signaling. The aim of this study was to explore whether the antioxidant effect of H2S is mediated by p66Shc. Application of exogenous H2S with its donor, NaHS, or overexpression of its generating enzyme, cystathionine β-synthase, induced sulfhydration of p66Shc, but inhibited its phosphorylation caused by H2O2/D-galactose in SH-SY5Y cells or in the mice cortex. H2S also decreased mitochondrial ROS production and protected neuronal cells against stress-induced senescence. PKCβII and PP2A are the two key proteins to regulate p66Shc phosphorylation. Although H2S failed to affect the activities of these two proteins, it disrupted their association. Cysteine-59 resides in proximity to serine-36, the phosphorylation site of p66Shc. The C59S mutant attenuated the above-described biological function of H2S. We revealed a novel mechanism for the antioxidant effect of H2S and its role in oxidative stress-related diseases.

H2S inhibits mitochondrial ROS production via the sulfhydration of Cys-59 residue, which in turn, prevents the phosphorylation of p66Shc.

Does pulmonary artery pulsatility index predict right ventricular failure after left ventricular assist device implantation?

Right ventricular failure (RVF) is a major cause of morbidity and mortality after left ventricular assist device (LVAD) implantation. The pulmonary artery pulsatility index (PAPi) is a novel hemodynamic index that predicts RVF in the setting of myocardial infarction, although it has not been shown to predict RVF after LVAD implantation. We performed a retrospective, single-center analysis to examine the utility of the PAPi in predicting RVF and RV assist device (RVAD) implantation in 85 continuous-flow LVAD recipients. We performed a multivariate logistic regression analysis incorporating previously identified predictors of RVF after LVAD placement, including clinical and echocardiographic variables, to determine the independent effect of PAPi in predicting RVF or RVAD after LVAD placement. In this cohort, the mean PAPi was 3.4 with a standard deviation of 2.9. RVF occurred in 33% of patients, and 11% required a RVAD. Multivariate analysis, adjusting for age, blood urea nitrogen (BUN), and Interagency Registry for Mechanically Assisted Circulatory Support profile, revealed that higher PAPi was independently associated with a reduced risk of RVAD placement (odds ratio [OR], 0.30; 95% confidence interval [CI], 0.07-0.89). This relationship did not change significantly when echocardiographic measures were added to the analysis. Stratifying the analysis by the presence of inotropes during catheterization revealed that PAPi was more predictive of RVAD requirement when measured on inotropes (OR, 0.21; 95% CI, 0.02-0.97) than without (OR, 0.49; 95% CI, 0.01-1.94). Furthermore, time from catheterization to LVAD did not significantly affect the predictive value of the PAPi (maximum time, 6 months). Receiver operating characteristic curve analysis revealed that optimal sensitivity and specificity were achieved using a PAPi threshold of 2.0.

In LVAD recipients, the PAPi is an independent predictor of RVF and the need for RVAD support after LVAD implantation. This index appears more predictive in patients receiving inotropes and was not affected by time from catheterization to LVAD in our cohort.

Is extracellular matrix metalloproteinase inducer associated with severity of brain oedema following experimental subarachnoid haemorrhage in rats?

Brain oedema is a major cause of clinical deterioration and death following brain trauma; cellular mechanisms involved in its development remain elusive. This study investigated the role of extracellular matrix metalloproteinase inducer (EMMPRIN) in brain oedema. The monofilament puncture model was used to induce subarachnoid haemorrhage. Adult male Sprague-Dawley rats were divided into five groups (n = 20 per group): sham-operated, sacrificed immediately after surgery (sham group); sacrificed 12, 24 or 72 h after subarachnoid haemorrhage induction (SAH-12, SAH-24 and SAH-72 groups, respectively); treated with EMMPRIN inhibitor immediately after subarachnoid haemorrhage, sacrificed at 24 h (SAH-inhibition group). Mean brain water content, and EMMPRIN mRNA and protein levels, were determined. Compared with the sham group, mean brain water content, EMMPRIN mRNA and protein levels in the SAH-12, SAH-24 and SAH-72 groups increased rapidly and significantly (maximal at 24 h). EMMPRIN inhibition significantly reduced mean brain water content and EMMPRIN mRNA and protein levels in the SAH-inhibition group, compared with the SAH-24 group.

EMMPRIN upregulation may be important in the formation of brain oedema; inhibition of EMMPRIN activity may provide a potential approach to reduce oedema after subarachnoid haemorrhage.

Is the early asthmatic response associated with glycolysis , calcium binding and mitochondria activity as revealed by proteomic analysis in rats?

The inhalation of allergens by allergic asthmatics results in the early asthmatic response (EAR), which is characterized by acute airway obstruction beginning within a few minutes. The EAR is the earliest indicator of the pathological progression of allergic asthma. Because the molecular mechanism underlying the EAR is not fully defined, this study will contribute to a better understanding of asthma. In order to gain insight into the molecular basis of the EAR, we examined changes in protein expression patterns in the lung tissue of asthmatic rats during the EAR using 2-DE/MS-based proteomic techniques. Bioinformatic analysis of the proteomic data was then performed using PPI Spider and KEGG Spider to investigate the underlying molecular mechanism. In total, 44 differentially expressed protein spots were detected in the 2-DE gels. Of these 44 protein spots, 42 corresponded to 36 unique proteins successfully identified using mass spectrometry. During subsequent bioinformatic analysis, the gene ontology classification, the protein-protein interaction networking and the biological pathway exploration demonstrated that the identified proteins were mainly involved in glycolysis, calcium binding and mitochondrial activity. Using western blot and semi-quantitative RT-PCR, we confirmed the changes in expression of five selected proteins, which further supports our proteomic and bioinformatic analyses.

Our results reveal that the allergen-induced EAR in asthmatic rats is associated with glycolysis, calcium binding and mitochondrial activity, which could establish a functional network in which calcium binding may play a central role in promoting the progression of asthma.

Is gender-related outcome difference related to course of sepsis on mixed ICUs : a prospective , observational clinical study?

Impact of gender on severe infections is in highly controversial discussion with natural survival advantage of females described in animal studies but contradictory to those described human data. This study aims to describe the impact of gender on outcome in mixed intensive care units (ICUs) with a special focus on sepsis. We performed a prospective, observational, clinical trial at Charité University Hospital in Berlin, Germany. Over a period of 180 days, patients were screened, undergoing care in three mainly surgical ICUs. In total, 709 adults were included in the analysis, comprising the main population ([female] n = 309, [male] n = 400) including 327 as the sepsis subgroup ([female] n = 130, [male] n = 197). Basic characteristics differed between genders in terms of age, lifestyle factors, comorbidities, and SOFA-score (Sequential Organ Failure Assessment). Quality and quantity of antibiotic therapy in means of antibiotic-free days, daily antibiotic use, daily costs of antibiotics, time to antibiotics, and guideline adherence did not differ between genders. ICU mortality was comparable in the main population ([female] 10.7% versus [male] 9.0%; P = 0.523), but differed significantly in sepsis patients with [female] 23.1% versus [male] 13.7% (P = 0.037). This was confirmed in multivariate regression analysis with OR = 1.966 (95% CI, 1.045 to 3.701; P = 0.036) for females compared with males.

No differences in patients' outcome were noted related to gender aspects in mainly surgical ICUs. However, for patients with sepsis, an increase of mortality is related to the female sex.

Is breast-feeding a protective factor for infectious diseases?

Case-control study. Retrospective study during 1998. Primary care. Teaching health center. Population of 22,500 inhabitants. We consider an alpha risk of .05 and a beta risk of .20, unilateral hypothesis. We estimated a sample of 48 children in the "case" group and 144 in the "control" group. Random sample. Cases were children with 3 or more acute respiratory or digestive infectious episodes during the first year of age. Controls were children with less than 3 episodes. We analyze exposure to breast feeding (BF) and we consider it if the child had at least 3 months of BF. In other case we define as a non-exposure child. We have calculated odds ratio (OR) and its 95% confidence intervals. 60.4% of the children were BF after birth. But only 28.5% lasted 3 months, and only 13.9% lasted 1 year. The median of BF was 3.3 months (SD 4.2). 79.2% of the children have a respiratory infectious disease in the first year and 14.6% diarrhea. In children less than 1 year old the OR was 0.81 (95% CI, 0.40-1.63). In the period 3-6 months old we have found an OR of 0.346 (95% CI 0.161-0.744).

In children aged less than 1 year, we have not found protective effects of breast-feeding against acute respiratory infectious diseases. Only in the period 3-6 months old, we have found that BF protects against these diseases.

Are super-races likely to dominate a fungal population within a life time of a perennial crop plantation of cultivar mixtures : a simulation study?

Deployment of cultivars with different resistance in mixtures is one means to manage plant diseases and prolong the life of resistance genes. One major concern in adopting mixtures is the development of 'super-races' that can overcome many resistance genes present in the mixture. A stochastic simulation model was developed to study the dynamics of virulence alleles in two-cultivar mixtures of perennial crops, focusing on the effects of cost of virulence and pathogen reproduction mechanism. The simulated mechanism of virulence has characteristics of both major and minor genes. Random genetic drift due to repeated population crashes during the overwintering phase led to fixation of a single fungal genotype (in terms of its virulence), often within 100 seasons. Overall, cost of virulence is most important in determining the virulence dynamics under the present model formulation. With cost of virulence incorporated, nearly all simulation runs ended up with a single fungal genotype that can infect only one of the two cultivars. In absence of cost of virulence, most of the simulation runs ended up with fungal genotypes that can infect both host cultivars but in many cases do not contain the maximum possible number of virulence alleles due to random drift. A minimum of 20% sexual reproduction between strains from different cultivars is necessary to ensure that the final fixed strains are able to infect both cultivars. Although the number of virulence alleles in the final genotype and the time to fixation are affected by simulation factors, most of the variability was among replicate simulation runs (i.e. stochastic in nature). The time to fixation is generally long relative to cropping cycles.

A single fungal genotype will dominate a population due to the bottleneck in overwintering with cost of virulence primarily determining whether the dominant genotype can infect both cultivars. However, the dominant genotype is unlikely to accumulate all the virulence alleles due to genetic drift. The risk of emergence and spread of super-races is insufficiently great to prevent the use of cultivar mixtures of perennial crops as a means to reduce disease development provided that host resistance structure in mixtures is altered every cropping cycle.

Is sARS-CoV envelope protein palmitoylation or nucleocapid association required for promoting virus-like particle production?

Coronavirus membrane (M) proteins are capable of interacting with nucleocapsid (N) and envelope (E) proteins. Severe acute respiratory syndrome coronavirus (SARS-CoV) M co-expression with either N or E is sufficient for producing virus-like particles (VLPs), although at a lower level compared to M, N and E co-expression. Whether E can release from cells or E/N interaction exists so as to contribute to enhanced VLP production is unknown. It also remains to be determined whether E palmitoylation or disulfide bond formation plays a role in SARS-CoV virus assembly. SARS-CoV N is released from cells through an association with E protein-containing vesicles. Further analysis suggests that domains involved in E/N interaction are largely located in both carboxyl-terminal regions. Changing all three E cysteine residues to alanines did not exert negative effects on E release, E association with N, or E enhancement of VLP production, suggesting that E palmitoylation modification or disulfide bond formation is not required for SARS-CoV virus assembly. We found that removal of the last E carboxyl-terminal residue markedly affected E release, N association, and VLP incorporation, but did not significantly compromise the contribution of E to efficient VLP production.

The independence of the SARS-CoV E enhancement effect on VLP production from its viral packaging capacity suggests a distinct SARS-CoV E role in virus assembly.

Is inducible costimulator ( ICOS ) a marker for highly suppressive antigen-specific T cells sharing features of TH17/TH1 and regulatory T cells?

CD4(+)CD25(+) regulatory T (Treg) cells are involved in the downmodulation of numerous immune responses to pathogens, tumors, or allergens. In this study, we further characterized the nature of Treg cells that control skin inflammatory reactions to haptens. In a model of contact hypersensitivity to 2,4-dinitro-fluorobenzene, we have investigated the phenotype, the specificity, and the origin of Treg cells that modulate the priming of effector CD8(+) T cells responsible for the development of the pathology. 2,4-Dinitrofluorobenzene immunization induced a population of CD4(+)CD25(+) Treg cells that controlled CD8(+) T-cell effector responses in a hapten-specific manner in vivo. High levels of inducible costimulator (ICOS) expression defined a population of CD4(+)CD25(+)FoxP3(+) (forkhead box protein 3) Treg cells that presented superior suppressive activity. Importantly, ICOS(+) Treg cells were distinguishable from all other FoxP3(+) Treg cells by the expression of IL-10, IL-17, and IFN-gamma. Hapten-specific Treg cells proliferating in response to their cognate antigen in vivo predominantly displayed a CD25(+)FoxP3(+)ICOS(+) phenotype. By using reporter mice, we showed that ICOS(+) Treg cells derived from the expansion of natural CD4(+)FoxP3(+) Treg cells rather than generation of adaptive Treg cells. Furthermore, the generation of ICOS(+) Treg cells depended on innate cells rather than the effector CD8(+) T-cell population.

Taken together, our data show that a population of CD4(+)CD25(+)FoxP3(+) T cells upregulates ICOS on in vivo sensitization and specifically suppresses hapten-reactive CD8(+) T cells both in vivo and in vitro.

Is intrahepatic natural killer cell activation , but not function , associated with HBsAg levels in patients with HBeAg-negative chronic hepatitis B?

Natural killer (NK) cells play an important role in the immune response to viruses. As the hepatitis B virus (HBV) replicates in hepatocytes, examination of the liver of chronic hepatitis B (CHB) patients is crucial to better understand the role of NK cells in HBV. HBeAg-negative CHB differs in many aspects from HBeAg-positive CHB, and until now little is known about the intrahepatic NK cell response in HBeAg-negative patients. Intrahepatic immune control might be different in HBeAg-negative as compared with HBeAg-positive patients. Liver NK cells were investigated in 21 HBeAg-positive and 35 HBeAg-negative CHB patients. Biopsy specimens were processed for routine histopathology and staging according to Ishak scores. Intrahepatic and blood NK cell frequencies, activation status and function of NK cells were analysed by flow cytometry. In HBeAg-negative CHB patients, compared to blood, liver NK cells displayed a more activated phenotype and stimulation further increased the activation status, but production of IFN-γ was markedly less. There was no difference with HBeAg-positive CHB. Only in HBeAg-negative CHB, but not in HBeAg-positive CHB, NK cell activation was inversely correlated with HBsAg levels.

The present study indicates that liver NK cells of CHB have a higher activation status compared to blood. However, they are not capable to increase cytokine production above levels reached by activated blood NK cells. In HBeAg-negative CHB, the levels of HBsAg may contribute to the incapacity of activated liver NK cells to increase cytokine production.

Cefuroxime for empiric treatment of community-acquired pneumococcal pneumonia: is there a generation gap?

Streptococcus pneumoniae infection is an important cause of morbidity and mortality. The recommendations to use expanded-spectrum beta-lactam drugs for patients with community-acquired pneumonia derived from the growing prevalence of penicillin-resistant pneumococci. Controversy exists regarding the use of second generation cephalosporins for empirical treatment of community-acquired pneumonia. In a retrospective study, 31 adult patients with pneumococcal pneumonia and bacteremia caused by S. pneumoniae that was intermediately resistant to penicillin were compared with 31 control patients with similar infection caused by penicillin-susceptible pneumococci. All patients were treated empirically with cefuroxime, alone or in combination with other antibiotics. Morbidity and mortality were studied. All unsusceptible pneumococci isolates were intermediately resistant to penicillin. No cases of fully resistant pneumococci were isolated from blood cultures in our hospital. Two factors were significantly associated with non-susceptibility to penicillin: hematologic malignancy and immunosuppression. No significant difference in morbidity or mortality was detected between the 2 groups, and penicillin minimum inhibitory concentration was not found to be a factor associated with mortality.

Patients with pneumococcal pneumonia caused by intermediately resistant pneumococci can be empirically treated with cefuroxime. In regions where fully resistant pneumococci are rare, the use of a second generation cephalosporin for empiric treatment of community-acquired pneumonia may be appropriate.

Does diastolic and systolic right ventricular dysfunction precede left ventricular dysfunction in patients paced from right ventricular apex?

Cardiac dysfunction after right ventricular (RV) apical pacing is well known but its extent, time frame of appearance and individual effect on left ventricular (LV), RV systolic and diastolic parameters has not evaluated in a systematic fashion. Patients with symptomatic bradycardia and ACC-AHA Class I indication for permanent pacemaker implantation (PPI) were implanted a single chamber (VVI) pacemaker. They were followed prospectively by echocardiographic examination which was done at baseline, 1 week, 1 month and 6 months after implantation. Parameters observed were chamber dimensions (M-line), chamber volumes, cardiac output (modified Simpson's method), systolic functions (ejection fraction, pre-ejection period, ejection time and ratio) and diastolic functions( isovolumic relaxation time & deceleration time) of left and right heart. Forty eight consecutive patients (mean age 65.6+/-11.8 yrs, 66.7% males, mean EF 61.82+/-10.36%) implanted a VVI pacemaker were enrolled in this study. The first significant change to appear in cardiac function after VVI pacing was in diastolic properties of RV as shown by increase in RV isovolumic relaxation time (IVRT) from 65.89+/-15.93 to 76.58+/-17.00 ms,(p<0.001) at 1week and RV deceleration time (DT) from 133.84+/-38.13 to 153.09+/-31.41 ms, (p=0.02) at 1 month. Increase in RV internal dimension (RVID) from 1.26+/-0.41 to 1.44+/-0.44, (p<0.05) was also noticed at 1 week. The LV diastolic parameters were significantly altered after 1 month with increase in LV-IVRT from 92.36+/-21.47 to 117.24+/-27.21ms, (p<0.001) and increase in LV DT from 147.56+/-31.84 to 189.27+/-28.49ms,(p<0.01). This was followed by LV systolic abnormality which appeared at 6 months with an increase in LVPEP from 100.33+/-14.43 to 118.41+/-21.34ms, (p<0.001) and increase in LVPEP/LVET ratio from 0.34+/-0.46 to 0.44+/-0.10, (p<0.001)]. The reduction in LV EF was manifested at 6 months falling from 61.82+/-10.36% to52.52+/-12.11%, (p<0.05) without any significant change in the resting cardiac output.

The present study shows that dysfunction of right ventricle is the first abnormality that occurs in VVI paced patients, which manifests by 1 week followed by LV dysfunction which starts appearing by 1 month and the diastolic dysfunctions precede the systolic dysfunction in both ventricles.

Are mitochondrial and oxidative stress genes differentially expressed in neutrophils of sJIA patients treated with tocilizumab : a pilot microarray study?

Various pathways involved in the pathogenesis of sJIA have been identified through gene expression profiling in peripheral blood mononuclear cells (PBMC), but not in neutrophils. Since neutrophils are important in tissue damage during inflammation, and are elevated as part of the acute phase response, we hypothesised that neutrophil pathways could also be important in the pathogenesis of sJIA. We therefore studied the gene profile in both PBMC and neutrophils of sJIA patients treated with tocilizumab. We studied the transcriptomes of peripheral blood mononuclear cells (PBMC) and neutrophils from eight paired samples obtained from 4 sJIA patients taken before and after treatment, selected on the basis that they achieved ACR90 responses within 12 weeks of therapy initiation with tocilizumab. RNA was extracted and gene expression profiling was performed using Affymetrix GeneChip Human Genome U133 Plus 2.0 microarray platform. A longitudinal analysis using paired t-test (p < 0.05 and FC ≥ 1.5) was applied to identify differentially expressed genes (DEGs) between the two time points followed by ingenuity pathway analysis. Gene Set Enrichment Analysis (GSEA) and quantitative real-time PCR were then performed to verify the microarray results. Gene ontology analysis in neutrophils revealed that response to tocilizumab significantly altered genes regulating mitochondrial dysfunction and oxidative stress (p = 4.6E-05). This was independently verified with GSEA, by identifying a set of oxidative genes whose expression correlated with response to tocilizumab. In PBMC, treatment of sJIA with tocilizumab appeared to affect genes in Oncostatin M signalling and B cell pathways.

For the first time we demonstrate that neutrophils from sJIA patients responding to tocilizumab showed significantly different changes in gene expression. These data could highlight the importance of mitochondrial genes that modulate oxidative stress in the pathogenesis of sJIA.

Does ursodeoxycholic acid decrease sodium-glucose cotransporter ( SGLT2 ) expression and oxidative stress in the kidney of diabetic rats?

Oxidative stress has been associated with diabetic complications like nephropathies. Recent studies indicate that ursodeoxycholic acid (UDCA) may be beneficial preventing diabetes-induced oxidative stress and secondary complications. Thus, we study if the UDCA-treatment decreases the expression of sodium-glucose cotransporter (SGLT2) and the oxidative stress in the kidney of diabetic rats. The diabetes model was established by intraperitoneal injection of streptozotocin (50mg/kg). SGLT2 expression was evaluated by western blot and RT-PCR. Oxidative stress was assessed by catalase (CAT), glutathione peroxidase (GPx) and superoxide dismutase activities (SOD) and immunohistochemical analysis of 3-nitrotyrosine (3-NT). Streptozotocin-induced diabetes caused hyperglycemia and lower body weight. The SGLT2 expression and mRNA levels increased in cortex of kidney from diabetic rats. The CAT activity decreased in cortex and medulla from diabetic rats, otherwise the GPx activity increased. Furthermore the 3-NT staining of kidney from diabetic rats increased compared to control rats. The UDCA treatment was able to decrease hyperglycemia and prevents the SGLT2 over-expression, restores the CAT and GPX activities and decreases 3-NT staining.

The UDCA treatment prevents the over-expression of SGLT2 and oxidative stress in kidney of diabetic rats.

Does quinoline derivative KB3-1 potentiate paclitaxel induced cytotoxicity and cycle arrest via multidrug resistance reversal in MES-SA/DX5 cancer cells?

The resistance to chemotherapeutic drugs is a major problem for successful cancer treatment. Multidrug resistance (MDR) phenotype is characterized by over-expression of P-glycoprotein (P-gp) on the cancer cell plasma membrane that extrudes drugs out of the cells. Therefore, novel MDR reversal agents are desirable for combination therapy to reduce MDR and enhance anti-tumor activity. Thus, the present study was aimed to evaluate the potent efficacy of novel quinoline derivative KB3-1 as a potent MDR-reversing agent for combined therapy with TAX. MDR reversing effect and TAX combined therapy were examined by Rhodamine accumulation and efflux assay and Confocal immunofluorescence microscopy, Western blotting, TUNEL assay, and cell cycle analysis. The discovery of quinoline-3-carboxylic acid [4-(2-[benzyl-3[-(3,4-dimethoxy-phenyl)-propionyl]-amino]-ethyl)-phenyl]-amide (KB3-1) as a novel MDR-reversal agent. KB3-1 significantly enhanced the accumulation and retention of a P-gp substrate, rhodamine-123 in the P-gp-expressing MES-SA/DX5 uterine sarcoma cells but not in the P-gp-negative MES-SA cells at non-toxic concentrations of 1 microM and 3 microM. Similarly, fluorescence microscopy observation revealed that KB3-1 reduced the effluxed rhodamine-123 expression on the membrane of MES-SA/DX5 cells. Consistent with decreased P-gp pumping activity, confocal microscopic observation revealed that KB3-1 effectively diminished the expression of P-gp in paclitaxel (TAX)-treated MES-SA/DX-5 cells. Furthermore, Western blotting confirmed that KB3-1 reduced P-gp expression and enhanced cytochrome C release and Bax expression in TAX treated MES-SA/DX-5 cells. In addition, KB3-1 enhanced TAX-induced apoptotic bodies in MES-SA/DX5 cells by TdT-mediated-dUTP nick-end labeling (TUNEL) staining assay aswell as potentiated TAX- induced cytotoxicity, G2/M phase arrest and sub-G1 apoptosis in MES-SA/DX5 cells but not in MES-SA cells. Interestingly, KB3-1 at 3 microM had comparable MDR-reversal activity to 10 microM verapamil, a well-known MDR- reversal agent.

KB3-1 can be a MDR-reversal drug candidate for combination chemotherapy with TAX.

Does detection of BRAF p.V600E Mutations in Melanoma by Immunohistochemistry have a Good Interobserver Reproducibility?

Assessment of BRAF p.V600E mutational status has become necessary for treatment of patients with metastatic melanoma. Detection of p.V600E mutation by immunohistochemistry was recently reported in several tumor types. To evaluate the interobserver reproducibility of BRAF p.V600E detection by immunohistochemistry in melanoma. Immunohistochemistry with VE1 antibody was performed on metastatic melanomas of 67 patients. Staining interpretation was performed on digital image virtual slides of tissue microarrays. The p.V600E status was determined by 7 pathologists from 3 European laboratories, blinded for other interpretations and for molecular biology results. Melanomas had p.V600E (n = 30), p.V600K (n = 4), p.K601E (n = 1), p.600-601delinsE (n = 1), or no p.V600 mutations (n = 31). Staining of p.V600E within mutated cells was cytoplasmic and diffuse, and for each case the staining on the 3 tissue microarray cores was similar. In 53 cases (79.1%) the 7 pathologists had perfect concordance. Agreement of interobserver reproducibility was almost perfect (κ = 0.81 [0.77-0.85]). Only 2 false-positive responses (0.9%) were obtained. The specificities reported were 100% for 5 pathologists (two of whom previously trained for p.V600E interpretation), and 97% for 2 untrained pathologists.

Detection of BRAF p.V600E mutation by immunohistochemistry in melanomas has an excellent interobserver reproducibility. Our results suggest that immunohistochemistry could be used as a first step for detection of BRAF p.V600E mutation, to identify patients with melanoma as candidates for BRAF inhibitors.

Does a YAP/TAZ-Regulated Molecular Signature be Associated with Oral Squamous Cell Carcinoma?

Oral squamous cell carcinoma (OSCC) is a prevalent form of cancer that develops from the epithelium of the oral cavity. OSCC is on the rise worldwide, and death rates associated with the disease are particularly high. Despite progress in understanding the mutational and expression landscape associated with OSCC, advances in deciphering these alterations for the development of therapeutic strategies have been limited. Further insight into the molecular cues that contribute to OSCC is therefore required. Here, we show that the transcriptional regulators YAP (YAP1) and TAZ (WWTR1), which are key effectors of the Hippo pathway, drive protumorigenic signals in OSCC. Regions of premalignant oral tissues exhibit aberrant nuclear YAP accumulation, suggesting that dysregulated YAP activity contributes to the onset of OSCC. Supporting this premise, we determined that nuclear YAP and TAZ activity drives OSCC cell proliferation, survival, and migration in vitro, and is required for OSCC tumor growth and metastasis in vivo. Global gene expression profiles associated with YAP and TAZ knockdown revealed changes in the control of gene expression implicated in protumorigenic signaling, including those required for cell cycle progression and survival. Notably, the transcriptional signature regulated by YAP and TAZ significantly correlates with gene expression changes occurring in human OSCCs identified by The Cancer Genome Atlas (TCGA), emphasizing a central role for YAP and TAZ in OSCC biology.

This study defines a YAP/TAZ-regulated transcriptional program in OSCC and reveals novel roles for nuclear YAP/TAZ activity in the onset and progression of this cancer.

Do Different Cyclooxygenase Inhibitors Impair Rotator Cuff Healing in a Rabbit Model?

The effect of selective and non-selective cyclooxygenase (COX) inhibitors on tendon healing was variable. The purpose of the study was to evaluate the influence of non-selective COX inhibitor, ibuprofen and flurbiprofen axetil and selective COX-2 inhibitor, celecoxib on the tendon healing process in a rabbit model. Ninety-six New Zealand rabbits were used as rotator cuff repair models. After surgery, they were divided randomly into four groups: ibuprofen (10 mg·kg-1·d-1), celecoxib (8 mg·kg-1·d-1), flurbiprofen axetil (2 mg·kg-1·d-1), and control group (blank group). All drugs were provided for 7 days. Rabbits in each group were sacrificed at 3, 6, and 12 weeks after tendon repair. Tendon biomechanical load failure tests were performed. The percentage of type I collagen on the bone tendon insertion was calculated by Picric acid Sirius red staining and image analysis. All data were compared among the four groups at the same time point. All data in each group were also compared across the different time points. Qualitative histological evaluation of the bone tendon insertion was also performed among groups. The load to failure increased significantly with time in each group. There were significantly lower failure loads in the celecoxib group than in the control group at 3 weeks (0.533 vs. 0.700, P = 0.002), 6 weeks (0.607 vs. 0.763, P = 0.01), and 12 weeks (0.660 vs. 0.803, P = 0.002), and significantly lower percentage of type I collagen at 3 weeks (11.5% vs. 27.6%, P = 0.001), 6 weeks (40.5% vs. 66.3%, P = 0.005), and 12 weeks (59.5% vs. 86.3%, P = 0.001). Flurbiprofen axetil showed significant differences at 3 weeks (failure load: 0.600 vs. 0.700, P = 0.024; percentage of type I collagen: 15.6% vs. 27.6%, P = 0.001), but no significant differences at 6 and 12 weeks comparing with control group, whereas the ibuprofen groups did not show any significant difference at each time point.

Nonsteroidal anti-inflammatory drugs can delay tendon healing in the early stage after rotator cuff repair. Compared with nonselective COX inhibitors, selective COX-2 inhibitors significantly impact tendon healing.

Is interferon-alpha-induced TRAIL on natural killer cells associated with control of hepatitis C virus infection?

Pegylated interferon-alpha (PEG-IFNalpha), in combination with ribavirin, controls hepatitis C virus (HCV) infection in approximately 50% of patients by mechanisms that are not completely understood. Beside a direct antiviral effect, different immunomodulatory effects have been discussed. Natural killer (NK) cells might be associated with control of HCV infection. We examined the effects of IFNalpha on human NK cells and its relevance to HCV infection. We performed gene expression profiling studies of NK cells following stimulation of peripheral blood mononuclear cells with IFNalpha. We evaluated IFNalpha-induced tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) expression using flow cytometry analyses of NK cells isolated from patients with acute or chronic hepatitis C that had received PEG-IFNalpha therapy. TRAIL was among the most up-regulated genes after IFNalpha stimulation of NK cells from healthy controls. After in vitro stimulation with IFNalpha, CD56(dim) NK cells from patients who had responded to PEG-IFNalpha therapy expressed higher levels of TRAIL than cells from patients with chronic hepatitis C. TRAIL expression, ex vivo, was inversely correlated with HCV-RNA levels during the early phase of PEG-IFNalpha therapy. In patients with acute hepatitis C, TRAIL expression on CD56(bright) NK cells increased significantly compared with cells from controls. In in vitro studies, IFNalpha-stimulated NK cells eliminated HCV-replicating hepatoma cells by a TRAIL-mediated mechanism.

IFNalpha-induced expression of TRAIL on NK cells is associated with control of HCV infection; these observations might account for the second-phase decline in HCV-RNA levels during PEG-IFNalpha therapy.

Are nEK1 and DYNC2H1 both involved in short rib polydactyly Majewski type but not in Beemer Langer cases?

The lethal short rib polydactyly syndromes (SRP type I-IV) are characterised by notably short ribs, short limbs, polydactyly, multiple anomalies of major organs, and autosomal recessive mode of inheritance. Among them, SRP type II (Majewski; MIM 263520) is characterised by short ovoid tibiae or tibial agenesis and is radiographically closely related to SRP type IV (Beemer-Langer; MIM 269860) which is distinguished by bowed radii and ulnae and relatively well tubulated tibiae. NEK1 mutations have been recently identified in SRP type II. Double heterozygosity for mutations in both NEK1 and DYNC2H1 in one SRP type II case supported possible digenic diallelic inheritance. The aim of this study was to screen DYNC2H1 and NEK1 in 13 SRP type II cases and seven SRP type IV cases. It was not possible to screen DYNC2H1 in two patients due to insufficient amount of DNA. The study identified homozygous NEK1 mutations in 5/13 SRP type II and compound heterozygous DYNC2H1 mutations in 4/12 cases. Finally, NEK1 and DYNC2H1 were excluded in 3/12 SRP type II and in all SRP type IV cases. The main difference between the mutation positive SRP type II group and the mutation negative SRP type II group was the presence of holoprosencephaly and polymycrogyria in the mutation negative group.

This study confirms that NEK1 is one gene causing SRP type II but also reports mutations in DYNC2H1, expanding the phenotypic spectrum of DYNC2H1 mutations. The exclusion of NEK1 and DYNC2H1 in 3/12 SRP type II and in all SRP type IV cases further support genetic heterogeneity.

Does non-dominant hand movement facilitate the frontal N30 somatosensory evoked potential?

Previous literature has shown that the frontal N30 is increased during movement of the hand contralateral to median nerve stimulation. This finding was a result of non-dominant left hand movement in right-handed participants. It is unclear however if the effect depends upon non-dominant hand movement or if this is a generalized phenomenon across the upper-limbs. This study tests the effect of dominant and non-dominant hand movement upon contralateral frontal and parietal somatosensory evoked potentials (SEPs) and further tests if this relationship persists in left hand dominant participants. Median nerve SEPs were elicited from the wrist contralateral to movement in both right hand and left hand dominant participants alternating the movement hand in separate blocks. Participants were required to volitionally squeeze (~20% of a maximal voluntary contraction) a pressure-sensitive bulb every ~3 seconds with the hand contralateral to median nerve stimulation. SEPs were continuously collected during the task and individual traces were grouped into time bins relative to movement according to the timing of components of the Bereitschaftspotential. SEPs were then averaged and quantified from both FCZ and CP3/4 scalp electrode sites during both the squeeze task and at rest. The N30 is facilitated during non-dominant hand movement in both right and left hand dominant individuals. There was no effect for dominant hand movement in either group.

N30 amplitude increase may be a result of altered sensory gating from motor areas known to be specifically active during non-dominant hand movement.

Do high expression of IL-17 and IL-17RE associate with poor prognosis of hepatocellular carcinoma?

Hepatocellular carcinoma (HCC) is a typical malignancy in a background of chronic inflammation. Th17 cells (a major source of IL-17) constitute crucial components of infiltrating inflammatory/immune cells in HCC and can amplify inflammatory response via binding to interleukin-17 receptor (IL-17R). Thus, we investigated the expression and clinical significance of IL-17 and IL-17 receptor family cytokines in HCC. The expression and prognostic value of IL-17 and IL-17R (A-E) were examined in 300 HCC patients after resection. Six Th17 associated cytokines in serum (n = 111) were quantified using enzyme-linked immunosorbent assays. Phenotypic features of IL-17+ CD4+ T cells were determined by flow cytometry analysis. High expression of intratumoral IL-17 and IL1-7RE were significantly associated with poorer survival (p = 0.016 and <0.001, respectively) and increased recurrence (both P < 0.001) of HCC patients. Moreover, intratumoral IL-17, individually or synergistically with IL-17RE, could predict HCC early recurrence and late recurrence. Also, peritumoral IL-17RE showed the prognostic ability in HCC (P < 0.001 for OS/TTR). Furthermore, expression levels of Th17 associated cytokines including IL-6, -22, -17R and TNF-α were increased in serum of HCC patients compared to haemangioma patients. Importantly, activated human hepatic stellate cells induced in vitro expansion of IL-17+ CD4+ T cells.

High expression of IL-17 and IL-17RE were promising predictors for poor outcome of HCC patients. The protumor power of IL-17 producing CD4+ T cells was probably involved in the crosstalk with different types of inflammatory/immune cells in HCC.

Is melanoma expression of matrix metalloproteinase-23 associated with blunted tumor immunity and poor responses to immunotherapy?

Matrix metalloproteinase-23 (MMP-23) can block the voltage-gated potassium channel Kv1.3, whose function is important for sustained Ca(2+) signaling during T cell activation. MMP-23 may also alter T cell activity and phenotype through cleavage of proteins affecting cytokine and chemokine signaling. We therefore tested the hypothesis that MMP-23 can negatively regulate the anti-tumor T cell response in human melanoma. We characterized MMP-23 expression in primary melanoma patients who received adjuvant immunotherapy. We examined the association of MMP-23 with the anti-tumor immune response - as assessed by the prevalence of tumor-infiltrating lymphocytes and Foxp3(+) regulatory T cells. Further, we examined the association between MMP-23 expression and response to immunotherapy. Considering also an in trans mechanism, we examined the association of melanoma MMP-23 and melanoma Kv1.3 expression. Our data revealed an inverse association between primary melanoma MMP-23 expression and the anti-tumor T cell response, as demonstrated by decreased tumor-infiltrating lymphocytes (TIL) (P = 0.05), in particular brisk TILs (P = 0.04), and a trend towards an increased proportion of immunosuppressive Foxp3(+) regulatory T cells (P = 0.07). High melanoma MMP-23 expression is also associated with recurrence in patients treated with immune biologics (P = 0.037) but not in those treated with vaccines (P = 0.64). Further, high melanoma MMP-23 expression is associated with shorter periods of progression-free survival for patients receiving immune biologics (P = 0.025). On the other hand, there is no relationship between melanoma MMP-23 and melanoma Kv1.3 expression (P = 0.27).

Our data support a role for MMP-23 as a potential immunosuppressive target in melanoma, as well as a possible biomarker for informing melanoma immunotherapies.

Does cerebrospinal fluid Presenilin-1 increase at asymptomatic stage in genetically determined Alzheimer 's disease?

Presenilin-1 (PS1), the active component of the intramembrane γ-secretase complex, can be detected as soluble heteromeric aggregates in cerebrospinal fluid (CSF). The aim of this study was to examine the different soluble PS1 complexes in the lumbar CSF (CSF-PS1) of individuals with Alzheimer's disease (AD), particularly in both symptomatic and asymptomatic genetically determined AD, in order to evaluate their potential as early biomarkers. Western blotting, differential centrifugation and co-immunoprecipitation served to determine and characterize CSF-PS1 complexes. We also monitored the assembly of soluble PS1 into complexes in a cell model, and the participation of Aβ in the dynamics and robustness of the stable PS1 complexes. There was an age-dependent increase in CSF-PS1 levels in cognitively normal controls, the different complexes represented in similar proportions. The total levels of CSF-PS1, and in particular the proportion of the stable 100-150 kDa complexes, increased in subjects with autosomal dominant AD that carried PSEN1 mutations (eight symptomatic and six asymptomatic ADAD) and in Down syndrome individuals (ten demented and ten non-demented DS), compared with age-matched controls (n = 23), even prior to the appearance of symptoms of dementia. The proportion of stable CSF-PS1 complexes also increased in sporadic AD (n = 13) and mild-cognitive impaired subjects (n = 12), relative to age-matched controls (n = 17). Co-immunoprecipitation demonstrated the association of Aβ oligomers with soluble PS1 complexes, particularly the stable complexes.

Our data suggest that CSF-PS1 complexes may be useful as an early biomarker for AD, reflecting the pathology at asymptomatic state.

Is bispectral index dynamics during propofol hypnosis similar in red-haired and dark-haired subjects?

We have previously shown that red hair is associated with increased desflurane requirement for immobility, compared with dark hair. The effect of red hair on IV anesthetic requirement remains unknown. We tested the hypothesis that the propofol concentration in the effect site associated with half maximal electroencephalogram response, Ce50, is at least 50% higher in subjects with red hair. We modeled the propofol concentration versus electroencephalogram response relationship using a 2-step approach in 29 healthy dark- and red-haired volunteers receiving a propofol infusion to produce loss of consciousness. Bispectral Index (BIS) was the measure of drug effect. The parameters of a 3-compartment pharmacokinetic model were fit to measured arterial propofol concentrations. The relationship between effect-site propofol concentration (Ce) and BIS was characterized using a sigmoid Emax model. Model performance and accuracy of the estimated parameters were evaluated using accepted metrics and bootstrap resampling. The effect of hair color on the Ce50 for BIS response in the final model was assessed using a threshold of 6.63 (P<0.01) in reduction of -2 log likelihood. The influence of body weight on the model was also assessed. The inclusion of hair color as a model covariate did not improve either the pharmacokinetic or the pharmacodynamic model. A separate analysis for the dark- and red-haired subjects estimated a median (95% confidence interval) Ce50 BIS of 2.71 μg/mL (2.28-3.36 μg/mL) and 2.57 μg/mL (1.68-3.60 μg/mL), respectively. Body weight was a significant covariate for the CL1 and V1.

Red hair phenotype does not affect the pharmacokinetics or pharmacodynamics of propofol.

Does the natural history of duplex-detected stenosis after femoropopliteal endovascular therapy suggest questionable clinical utility of routine duplex surveillance?

Duplex ultrasound (DU) surveillance (DUS) criteria for vein graft stenosis and thresholds for reintervention are well established. The natural history of DU-detected stenosis and the threshold criteria for reintervention in patients undergoing endovascular therapy (EVT) of the femoropopliteal system have yet to be determined. We report an analysis of routine DUS after infrainguinal EVT. Consecutive patients undergoing EVT of the superficial femoral artery (SFA) or popliteal artery were prospectively enrolled in a DUS protocol (≤1 week after intervention, then at 3, 6, and 12 months thereafter). Peak systolic velocity (PSV) and velocity ratio (Vr) were used to categorize the treated artery: normal was PSV <200 cm/s and Vr <2, moderate stenosis was PSV = 200-300 cm/s or Vr = 2-3, and severe stenosis was PSV >300 cm/s or Vr >3. Reinterventions were generally performed for persistent or recurrent symptoms, allowing us to analyze the natural history of DU-detected lesions and to perform sensitivity and specificity analysis for DUS criteria predictive of failure. Ninety-four limbs (85 patients) underwent EVT for SFA-popliteal disease and were prospectively enrolled in a DUS protocol. The initial scans were normal in 61 limbs (65%), and serial DU results remained normal in 38 (62%). In 17 limbs (28%), progressive stenoses were detected during surveillance. The rate of thrombosis in this subgroup was 10%. Moderate stenoses were detected in 28 (30%) limbs at initial scans; of these, 39% resolved or stabilized, 47% progressed to severe, and occlusions developed in 14%. Five (5%) limbs harbored severe stenoses on initial scans, and 80% of lesions resolved or stabilized. Progression to occlusion occurred in one limb (20%). The last DUS showed 25 limbs harbored severe stenoses; of these, 13 (52%) were in symptomatic patients and thus required reintervention regardless of DU findings. Eleven limbs (11%) eventually occluded. Sensitivity and specificity of DUS to predict occlusion were 88% and 60%, respectively.

DUS does not reliably predict arterial occlusion after EVT. Stenosis after EVT appears to have a different natural history than restenosis after vein graft bypass. EVT patients are more likely to have severe stenosis when they present with recurrent symptoms, in contrast to vein graft patients, who commonly have occluded grafts when they present with recurrent symptoms. The potential impact of routine DU-directed reintervention in patients after EVT is questionable. The natural history of DU-detected stenosis after femoropopliteal endovascular therapy suggests questionable clinical utility of routine DUS.

Does nutritional screening with Subjective Global Assessment predict hospital stay in patients with digestive diseases?

Nutritional status is an important factor that determines hospital stay, and the Subjective Global Assessment (SGA) is a candidate tool for nutritional screening on admission. However, the significance of the SGA has not been evaluated well in the ward for digestive diseases. We conducted the present study to test whether the SGA predicts hospital stay of these patients. Two hundred sixty-two patients with digestive diseases were consecutively enrolled between July 2004 and April 2005. They consisted of 145 males and 117 females and included 110 patients with cancer. Disease category was gastrointestinal in 94, hepatic in 111, and biliary/pancreatic in 57. The SGA was performed by a certified dietician. Effects of SGA and other nutritional parameters on hospital stay were examined by simple and multiple regression analysis. Among tested variables, simple regression analysis identified the SGA, disease category, presence of malignancy, serum albumin level, percent triceps skinfold thickness, and percent arm muscle circumference as significant predictive parameters for hospital stay. Multiple regression analysis revealed that the SGA had the best predictive power, followed by the presence of malignancy and disease category.

The SGA is a simple and reliable predictor for hospital stay in patients with digestive diseases.

Does anandamide mediate hyperdynamic circulation in cirrhotic rats via CB ( 1 ) and VR ( 1 ) receptors?

Hyperdynamic circulation and mesenteric hyperaemia are found in cirrhosis. To delineate the role of endocannabinoids in these changes, we examined the cardiovascular effects of anandamide, AM251 (CB(1) antagonist), AM630 (CB(2) antagonist) and capsazepine (VR1 antagonist), in a rat model of cirrhosis. Cirrhosis was induced by bile duct ligation. Controls underwent sham operation. Four weeks later, diameters of mesenteric arteriole and venule (intravital microscopy), arterial pressure, cardiac output, systemic vascular resistance and superior mesenteric artery (SMA) flow were measured after anandamide, AM251 (with or without anandamide), AM630 and capsazepine administration. CB(1), CB(2) and VR1 receptor expression in SMA was assessed by western blot and RT-PCR. Anandamide increased mesenteric vessel diameter and flow, and cardiac output in cirrhotic rats, but did not affect controls. Anandamide induced a triphasic arterial pressure response in controls, but this pattern differed markedly in cirrhotic rats. Pre-administration of AM251 blocked the effects of anandamide. AM251 (without anandamide) increased arterial pressure and systemic vascular resistance, constricted mesenteric arterioles, decreased SMA flow and changed cardiac output in a time-dependent fashion in cirrhotic rats. Capsazepine decreased cardiac output and mesenteric arteriolar diameter and flow, and increased systemic vascular resistance in cirrhotic rats, but lacked effect in controls. Expression of CB(1) and VR1 receptor proteins were increased in cirrhotic rats. AM630 did not affect any cardiovascular parameter in either group.

These data suggest that endocannabinoids contribute to hyperdynamic circulation and mesenteric hyperaemia in cirrhosis, via CB(1)- and VR1-mediated mechanisms.

Does neural tissue engineering scaffold with sustained RAPA release relieve neuropathic pain in rats?

To investigate the effect of locally slow-released rapamycin (RAPA) from bionic peripheral nerve stent to reduce the incidence of neuropathic pain or mitigate the degree of pain after nerve injury. We constructed a neural tissue engineering scaffold with sustained release of RAPA to repair 20mm defects in rat sciatic nerves. Four presurgical and postsurgical time windows were selected to monitor the changes in the expression of pain-related dorsal root ganglion (DRG) voltage-gated sodium channels 1.3 (Nav1.3), 1.7 (Nav1.7), and 1.8 (Nav1.8) through immunohistochemistry (IHC) and Western Blot, along with the observation of postsurgical pathological pain in rats by pain-related behavior approaches. Relatively small upregulation of DRG sodium channels was observed in the experimental group (RAPA+poly(lactic-co-glycolic acid) (PLGA)+stent) after surgery, along with low degrees of neuropathic pain and anxiety, which were similar to those in the Autologous nerve graft group.

Autoimmune inflammatory response plays a leading role in the occurrence of post-traumatic neuropathic pain, and that RAPA significantly inhibits the abnormal upregulation of sodium channels to reduce pain by alleviating inflammatory response.

Does delayed blastocyst development influence the outcome of frozen-thawed transfer cycles?

To compare the outcome of transfer of thawed blastocysts frozen on either day 5 or day 6 after in vitro fertilisation. Cohort observational study. Tertiary assisted conception unit in London, UK. Six hundred and forty-two consecutive nondonor programmed thawed blastocyst transfer (TBT) cycles. High-grade blastocysts were frozen on day 5 (n = 314) or day 6 (n = 328) after fertilisation using a slow-freezing protocol. Endometrial preparation was performed using estradiol valerate. Progesterone supplementation was commenced when the endometrial thickness had reached 7 mm or more. Frozen blastocysts were thawed on day 6 of progesterone supplementation and assessed immediately after thawing for survival, and after 3-4 hours for blastocoele re-expansion. Main outcome measures Thawed blastocyst survival and re-expansion rates, and pregnancy and live birth rates, per TBT. Thawed blastocyst survival and re-expansion rates were comparable between the day 5 and day 6 groups (87% versus 87%, P = 0.50 and 73% versus 71%, P = 0.35, respectively). The live birth rate was similar between the two groups (29% versus 28.5%, P = 0.93, respectively). After adjusting for confounding variables, the odds ratio (OR) of a live birth in cycles in which the thawed blastocysts were frozen on day 6 compared with day 5 was 1.23 [95% confidence interval (CI), 0.81-1.86, P = 0.34].

The pregnancy potential of high-grade blastocysts frozen on day 5 and day 6 after in vitro fertilisation and replaced in programmed TBT cycles is comparable.

Are follicular helper T cells responsible for IgE responses to Der p 1 following house dust mite sensitization in mice?

Th2 cells have long been considered responsible for the switching of B cells to production of IgE during cognate interaction, primarily due to their expression of CD40L and secretion of IL-4. This concept has been challenged by the more recent definition of follicular helper T cells (Tfh) as the key T cell subset in B cell isotype switching, due to their physical location at the boundary of T cell:B cell areas in lymphoid follicles and ability to express IL-4 and CD40L. To determine whether Tfh cells are responsible for IgE responses to Der p 1 allergen after house dust mite (HDM)-induced allergic sensitization. Mice deficient in Tfh cells were sensitized to HDM and Der p 1-specific IgE measured by ELISA. Mice with a mutation in T cell-expressed IL-6R were unable to expand Tfh populations after HDM sensitization, and their anti-Der p 1 IgE, IgG1 and total IgE responses were reduced by 80-90% compared with wild-type mice. These animals displayed unaltered lung Th2 and eosinophilic responses after intranasal HDM challenge and normal IL-4 production, but B cell infiltration of the airways was abrogated.

Our data indicate that Tfh cells are largely responsible for switching B cells to IgE synthesis, most likely via an IgG1(+) intermediate. However, Th2 cells are the major source of IL-4 during HDM sensitization and this might contribute to IgE synthesis at a stage distal to Tfh-mediated isotype switching. The IL-6/follicular helper T cell pathway is a potential therapeutic target in allergic disease.

Is reduced expression of endothelial connexins 43 and 37 in hypertensive rats rectified after 7-day carvedilol treatment?

The aim of this study was to clarify the response of endothelial connexins to hypertension and antihypertensive drugs. Rats remained normotensive (group 1, N = 10) or were made hypertensive using N(omega)-nitro-L-arginine methyl ester (L-NAME) (groups 2 to 4, N = 10/group). Seven weeks later groups 3 and 4 were respectively fed atenolol or carvedilol daily for 7 days and the aortic endothelial gap junctions were analyzed. In parallel the effects of atenolol, carvedilol, labetalol, vitamin C, and vitamin E on connexin43 (Cx43) in human umbilical vein endothelial cells (HUVEC) were compared. Endothelial Cx43 was reduced by 35% and Cx37 by 59% in hypertensive rats (P < .001). The reduction was recovered fully by carvedilol but only partially by atenolol (P < .05), although both drugs lowered the blood pressure to similar levels. Cx40 remained stationary in all groups. In HUVEC, carvedilol (10 microg/mL) increased Cx43 protein expression by >70% (P < .01), whereas other drugs had minimal effects. The upregulation by carvedilol was associated with increased transcripts and decreased proteolysis of Cx43.

The study showed that L-NAME-induced hypertension has differential effects on endothelial connexins, which respond variously to carvedilol and atenolol. In HUVEC carvedilol directly upregulates endothelial Cx43 and the effect is independent of its antioxidant activity.

Does abdominal shape of gastric cancer patients influence short-term surgical outcomes?

There is a prevailing belief that both obesity and abdominal shape influence abdominal accessibility, and thus affect short-term surgical outcomes of gastric cancer surgery. We measured the thickness of subcutaneous fat (SCF), abdominal anterior-posterior diameter (APD), transverse diameter (TD), and intra-abdominal fat volume (IFV) at the umbilicus level by using the abdominal CT scans of 291 gastric cancer patients who had undergone subtotal gastrectomy and D2 lymph node dissection. Clinicopathological factors including body mass index (BMI), APD, TD, IFV, and SCF and surgical outcomes, i.e., dissected lymph node number, morbidity, and mortality were analyzed. SCF thickness, APD, TD, IFV, and BMI mean values were 20.0 mm (range 2.0-64.0), 188.4 mm (range 128.0-332.0), 301.4 mm (range 160.0-651.0), 198.3 mm(2) (range 123.4-312.1), and 23.9 kg/m(2) (range 16.6-34.6), respectively. In male patients, APD was found to correlate with the number of retrieved lymph nodes (P = 0.045). Whereas in female patients, this was not the case (P = 0.093). Twenty-one patients experienced postoperative complications but no postoperative mortality occurred. Female patients who experienced postoperative complications had higher APD (32.9 +/- 10.0 mm versus 26.1 +/- 7.9 mm, P = 0.044) and BMI (27.3 +/- 4.1 kg/m(2) versus 24.3 +/- 3.5 kg/m(2), P = 0.049) values than those who did not.

We conclude that obesity and abdominal shape of gastric cancer patients both influence the short-term surgical outcomes of subtotal gastrectomy with D2 lymph node dissection.

Does resection margin influence survival after pancreatoduodenectomy for distal cholangiocarcinoma?

Distal cholangiocarcinoma remains a rare cancer associated with a dismal outcome. There is a lack of effective treatment options and where disease is amendable to resection, surgery affords the best potential for long-term survival. The aim of this study was to examine the survival outcomes and prognostic factors of patients undergoing pancreatoduodenectomy for distal cholangiocarcinoma. Between January 2004 to May 2016, patients who had undergone pancreatoduodenectomy with histologically proven distal cholangiocarcinoma were identified. Clinicopathologic data and survival outcomes were reported. Pancreatoduodenectomy alone was performed in 20 patients (71%) and eight patients (29%) required concomitant vascular resection. The major complication rate was 43% (n = 12). Nineteen patients (68%) had node positive disease. Eighteen patients (64%) had R0 resection. The median survival was 36 months (95%CI 9.7 to 63.8) and 5-year survival rate was 24%. Univariate analysis identified ASA (P < 0.001), tumor grade (P = 0.009) and margin status (P = 0.042) as prognostic factors associated with survival.

Long-term survival may be achieved in selected patients undergoing pancreatoduodenectomy for distal cholangiocarcinoma, especially in patients who achieved an R0 resection.

Does initial bradykinin trigger calcium-induced calcium release in C6 glioma cells and its significance?

To investigate the underlying mechanism for the selective modulation of the permeability of blood-tumor barrier (BTB) by small dose of bradykinin (BK). C6 glioma cells were treated with BK, and changes of intracellular nitric oxide (NO) and intracellular calcium level were measured with fluorescent spectrophotometer. The initial application of BK easily triggered extracellular calcium influx, which resulted in intracellular calcium store release in C6 glioma cells. The above mechanism was also named ryanodine mediated calcium induced calcium release (CICR). We also detected a long-lasting intracellular NO elevation in C6 glioma cells upon BK treatment. Further study showed that ryanodine mediated CICR contributed greatly to the secondary NO elevation induced by BK treatment.

These results suggested that BK triggered CICR in C6 glioma cells and the associated NO generation might be the underlying mechanism for the selective modulation of BTB permeability by BK.

Do positive short-term subjective effect of sports drink supplementation during recovery?

The aim of this study was to investigate the effects of a naturally composed sports drink containing proteins and carbohydrates used during recovery in competitive badminton players. The hypothesis was that the use of a recovery drink would lead to positive subjective effects, enhanced physical performance and less signs of overtraining. During an in-door season 18 badminton players were instructed to drink at least 250 mL of a given sports drink immediately after each training or playing session. The study design was prospective double blind crossover with one active drink and one placebo. The active drink was based on natural products containing whey and orange juice, and the placebo was made of diluted apple juice. Evaluation of effects was done with laboratory tests, self-registered values and field tests. The players perceived statistically significant short-term subjective positive effects after using the active drink, compared with after using placebo. The blood hemoglobin concentration was also higher after the period with active drink. There were no other differences concerning other laboratory tests (leg strength, endurance, body fat percent, lean arm and leg masses), self-registered values (body weight, pulse, training amount and intensity) or field tests (speed, explosive effort, grip strength, endurance and POMS) between the periods with the different sports drinks.

Supplementation with a sports drink during recovery showed a significant short-term subjective positive effect compared with placebo. However, no effects were seen on physical performance or signs of overtraining.

Do long-term performance of the attain model 4193 left ventricular lead?

Cardiac resynchronization therapy (CRT) has proven to be a valuable therapy addition for patients with drug-refractory heart failure and a ventricular conduction delay. Delivery of CRT is dependent upon the successful implantation and chronic performance of a left ventricular (LV) pacing lead. This study assessed the long-term electrical performance and safety of a steroid-eluting, transvenous, over-the-wire, cardiac vein pacing lead. The Attain Model 4193 LV lead (Medtronic, Inc, Minneapolis MN, USA) was successfully implanted in 1,070 patients with 286 patients completing 3 years of follow-up. Clinical data were collected at pre-implant, implant, and at 6-month intervals for 3 years. Over 3 years, the mean chronic pacing threshold ranged from 1.9 V to 2.1 V, the mean R-wave sensing amplitudes ranged between 13.6 mV and 15.0 mV, and the mean pacing impedance ranged between 562 ohms and 590 ohms. Additionally, the observed freedom from first post-implant LV-lead-related complications was 90.4%. Of 1,070 total patients, 82 experienced 89 LV-lead-related adverse events requiring invasive interventions or resulting in the termination of the CRT therapy. The LV lead was repositioned in 31 patients, replaced in 21 patients, and explanted/capped in four patients. There were no deaths related to the LV lead during implantation or during the follow-up period.

The data suggest that the 4193 LV lead is safe and effective over time. The LV lead electrical measurements remained stable through follow-up, demonstrating reliable long-term performance within the recommended value range at 36 months and had an acceptable complication rate.

Do hospital employee assault rates before and after enactment of the california hospital safety and security act?

This study examines changes in violent event rates to hospital employees before and after enactment of the California Hospital Safety and Security Act in 1995. We compared pre- and post-initiative employee assault rates in California (n = 116) emergency departments and psychiatric units with those in New Jersey (n = 50), where statewide workplace violence initiatives do not exist. Poisson regression with generalized estimating equations was used to compare assault rates between a 3-year pre-enactment period (1993-1995) and a 6-year post-enactment period (1996-2001) using New Jersey hospitals as a temporal control. Assault rates among emergency department employees decreased 48% in California post-enactment, compared with emergency department employee assault rates in New Jersey (rate ratio [RR] = 0.52, 95% confidence interval [CI]: 0.31, 0.90). Emergency department employee assault rates decreased in smaller facilities (RR = 0.46, 95% CI: 0.21, 0.96) and for-profit-controlled hospitals (RR = 0.39, 95% CI: 0.19, 0.79) post-enactment. Among psychiatric units, for-profit-controlled hospitals (RR = 0.41, 95% CI: 0.19, 0.85) and hospitals located in smaller communities (RR = 0.44, 95% CI: 0.21, 0.92) experienced decreased assault rates post-enactment.

Policy may be an effective method to increase safety to health care workers.

Do indications for internal cardioverter defibrillator implantation predict time to first shock and the modulating effect of beta-blockers?

Patients receive implantable cardioverter defibrillator (ICD) for varying indications. Whether these indications influence the time to first ICD shock is suspected but not confirmed. The modulating effect of beta-blockers on shock-free survival is not fully elucidated. A retrospective analysis of 230 consecutive patients (age 63 +/- 14 years, 79% men, 75% ischemic, 70% beta-blockers) implanted with an ICD was performed. Patients were divided into 4 groups depending on the ICD indication: groups A (secondary prevention of sudden death), B (left ventricular ejection fraction < or = 35% and positive electrophysiology study [EPS]), C (left ventricular ejection fraction < or = 35% and negative EPS or no EPS performed), and D (patients who did not meet inclusion criteria for groups A, B, or C). Time to shock was analyzed by the Kaplan-Meier method. During a mean follow-up of 489 +/- 280 days, 57 (24.7%) patients received 82 shocks (49% appropriate). The 1-year shock-free survival for patients in groups A, B, C, and D were 57%, 77%, 79%, and 91%, respectively (P = .03), for total shocks and 75%, 92%, 92%, and 100%, respectively (P = .007), for appropriate shocks. For patients in group A, the use of beta-blockers increased the 1-year shock-free survival from 48% to 61% for total shocks and from 65% to 79% for appropriate shocks.

Time to first shock is determined by the indication for ICD implantation and is not predicted by the results of EPS. Patients with secondary indications for ICD implantation are at highest risk of shocks and may deserve consideration for prophylactic antiarrhythmic drugs. beta-Blockers increase the time to first ICD shock in patients implanted for secondary prevention of sudden death.

Is the number of regular T cells and immature dendritic cells involved in mycosis fungoides linked to the tumor stage?

The balance between immune surveillance and immune escape determines the outcome of patients with primary mycosis fungoides (MF). FOXP3+ regulatory T cells (Tregs) and DC-SIGN+ immature dendritic cells (imDCs) play a central role in regulating the immune state in the progression of MF. However, whether the mechanisms used by these factors depend on MF stage is still underdetermined and even controversial. FOXP3+ Tregs and DC-SIGN+ imDCs were detected by immunohistochemical staining of formalin-fixed, paraffin-embedded specimens obtained from the lesion biopsies of 89 patients with MF, comprising 69 patients at the patch stage, 12 at the plaque stage, and 8 at the tumor stage. The number of FOXP3+ Tregs and DC-SIGN+ imDCs in each stage was counted and compared. The expression of FOXP3 and DC-SIGN varied with the MF stage. The number of cells expressing FOXP3 was higher at the patch and plaque stages than at the tumor stage (p < 0.05), but no significant difference was noted between the patch and plaque stages (p = 0.715). DC-SIGN expression increased continuously, concomitant with tumor progression, through the three stages (p < 0.05).

The predominant factor influencing the immune state is different for each MF stage. Therefore, therapeutic strategies that modulate the antitumor immune responses should be developed depending on MF progression.

Are hFE gene mutations associated with osteoarthritis in the index or middle finger metacarpophalangeal joints?

. To test the hypothesis that possession of either C282Y or H63D mutations in the HFE gene is associated with primary osteoarthritis (OA) in joints commonly affected in hemochromatotic arthropathy. HFE genotyping was performed in 87 patients with radiologically proven OA in 3 joint regions: index or middle finger metacarpophalangeal joints (MCP2,3; n = 52), elbow joints (n = 8), ankle, intertarsal or tarsometatarsal joints (ankle/IT/TMT; n = 27); and in 56 patients with radiologically proven OA in finger interphalangeal (IP) joints, but not MCP2,3 joints (IP OA control group). HFE mutation frequencies in these groups were also compared to those in a similar population (Busselton population control group). A statistically significant association between HFE mutations and OA was observed for the MCP2,3 joints (p = 0.0001) and the ankle/IT/TMT joint group (p = 0.002) as well as for the 3 joint regions collectively (p = 0.0001), but not for the elbow joints (p = 0.062). Comparison with the Busselton population controls showed similar statistically significant associations, except for the elbow and ankle/IT/TMT groups, where similar trends were observed.

HFE gene mutations are associated with OA in the MCP2,3 joints. These mutations may be markers for a polyarticular OA phenotype.

Does generalizability of clinical trial result for generalized anxiety disorder to community samples?

There has been little research on the generalizability of clinical trials for generalized anxiety disorder (GAD). The present study examines the generalizability of pharmacological and psychotherapy clinical trials' results of individuals with DSM-IV GAD to a large community sample. Data were drawn from the National Epidemiological Survey on Alcohol and Related Conditions (NESARC), a large national representative face-to-face sample of 43,093 adults of the United States population. We applied a standard set of eligibility criteria representative of GAD pharmacological and psychotherapy clinical trials to all adults with past 12 months GAD (n = 894), and to a subgroup of participants seeking treatment (n = 329). Our aim was to assess how many participants with GAD would fulfil typical eligibility criteria. We found that more than seven out of 10 participants with GAD were excluded by at least one criterion. In the subgroup of GAD participants who sought treatment, the exclusion rate by at least one criterion raised to more than eight out of 10 participants with GAD. For the overall sample and the treatment-seeking subsample, having a current depression was the criterion excluding the highest percentage of individuals. Having a lifetime history of bipolar disorder, a current significant medical condition, a current diagnosis of alcohol abuse or dependence, and a social or specific phobia also excluded a substantial proportion of individuals in both samples.

Clinical trials exclude a majority of adults with GAD. Clinical trials should carefully consider the impact of eligibility criteria on the generalizability of their results.

Does gamma-secretase inhibition combined with platinum compounds enhance cell death in a large subset of colorectal cancer cells?

Notch signalling is essential for the development and maintenance of the colonic epithelium. Its inhibition induces a differentiation phenotype in vivo and reduces adenomas in APCmin mice. Whether Notch signals are also required in colorectal cancer (CRC) has remained elusive. Therefore, 64 CRC cell lines were analysed for the occurrence of proteolytically processed, active Notch. 63 CRC lines contained a fragment with approximately the size of the Notch1 intracellular domain (NICD), which is required for signalling. Subsequent analyses with an antibody that specifically recognises the free Val1744 residue generated by gamma-secretase-mediated cleavage of Notch1 showed that a subset of CRC cells lacks this specific Val1744-NICD. Surprisingly, inhibition of Val1744-NICD signalling with different gamma-secretase inhibitors (GSI) did not lead to substantial effects on CRC cell line growth or survival. However, transient activation of Erk upon GSI treatment was detected. Since cisplatin relies on Erk activation for bioactivity in some cells, platinum compounds were tested together with GSI and enhanced cell killing in a subset of Val1744-NICD-positive CRC cell lines was detected. Erk inhibition ablated this combination effect.

We conclude that gamma-secretase inhibition results in activation of the MAP kinases Erk1/2 and, when used in conjunction, enhances cell death induced by platinum compounds in a large subset of colorectal cancer cell lines.Furthermore the activation of Erk appears to be of particular importance in mediating the enhanced effect seen, as its inhibition abrogates the observed phenomenon. These findings do not only highlight the importance of signalling pathway crosstalk but they may also suggest a new avenue of combination therapy for some colorectal cancers.

Does stimulation of serotonin2C receptors influence cocaine-seeking behavior in response to drug-associated stimuli in rats?

It has been suggested that the increase in serotonin transmission induced by indirect agonists such as fenfluramine and fluoxetine attenuates cue-elicited reinstatement of cocaine-seeking in rats through a 5-HT2C receptor-dependent mechanism. We investigated whether Ro 60-0175, a nonselective 5-HT2B-2C agonist, influences cue-elicited reinstatement of cocaine-seeking behavior. We evaluated the 5-HT2C receptor's role in Ro 60-0175 by studying its interaction with SB-242,084, a selective 5-HT2C antagonist. The study also explored whether Ro 60-0175 influences cue-elicited seeking behavior associated with sucrose, a highly palatable nutritive reinforcer. Different groups of free-feeding rats were trained to associate discriminative stimuli (SDs) with the availability of cocaine or a sucrose pellet or no-reward in two-lever operant cages. Cocaine and sucrose pellets were available under an FR1 schedule of reinforcement, and each reinforcer was followed by a 20-s timeout signaled by a cue light coming above the active lever. After extinction of reinforced responding in the absence of cue, the reinforcer-associated stimuli were reintroduced in reinstatement sessions in which reinforcers were withheld. Ro 60-0175, at IP doses from 0.1 to 1 mg/kg, dose-dependently reduced cocaine-seeking behavior, while 1 mg/kg had no such effect for the sucrose pellet. Pretreatment with 1 mg/kg SC SB-242,084 completely prevented the effect on cocaine-seeking behavior.

These findings, provided they can be extrapolated to abstinent human addicts, suggest therapeutic potential for the selective 5-HT2C agonist in preventing cue-controlled cocaine-seeking and relapse.

Can we improve the diagnosis of spondyloarthritis in patients with uncertain diagnosis?

Power Doppler ultrasound (PDUS) has proved to be a highly sensitive tool for assessing enthesitis in spondyloarthritis (SpA). In patients with a suspected SpA, diagnosis could be improved by detecting enthesitis with PDUS. To evaluate the performance of PDUS for the diagnosis of SpA alone or combined with other clinical, laboratory and imaging findings in patients consulting for a suspected SpA. Prospective, multicenter French cohort study (Boulogne-Billancourt, Brest, Caen, Grenoble, Marseille and Nancy). Outpatients consulting for symptoms suggestive of SpA (inflammatory back pain [IBP], arthritis or inflammatory arthralgia [IA], enthesitis or dactylitis [ED], HLA-B27 positive uveitis [B27+U], familiarity for SpA [Fam]) were recruited and followed up for at least 2 years. Sample size was set to 500 patients (for estimated prevalence of SpA of 30±5% after 2 years). At baseline, patients were submitted to standardized physical examination, pelvic X-ray, sacroiliac joints magnetic resonance imaging (MRI), HLA-B typing, and other tests judged useful for diagnosis. For each patient, a blinded PDUS examination of 14 enthesitic sites was performed at baseline and at years 1 and 2. Patients were planned to be followed during 5 years. The diagnosis of SpA ascertained by an experts' committee, blind to PDUS results, after at least 2 years of follow-up, with a revaluation of doubtful patients at 5 years will be used as gold standard for evaluating the diagnostic performance of PDUS and the best diagnostic procedure by combining PDUS, clinical symptoms and other tests. Between January 2005 and September 2007, 489 patients were included (96% of the target population). Nineteen patients (0.2%) retired their informed consensus or were lost to follow-up immediately after their inclusion. At baseline, mean age of the 470 remaining patients was 40 years, mean duration of symptoms was 6.1 years; 42% of them were HLA-B27+ and 63% were female. Primary inclusion criterion was IBP in 53%, IA in 27%, ED in 9%, B27+U in 8% and Fam in 4%. Follow-up is still ongoing.

We have set up a unique diagnostic cohort which includes the entire spectrum of SpA manifestations. By using PDUS we expected to improve the diagnostic procedure of SpA.

Does novel evidence demonstrate that epithelial-mesenchymal transition contributes to nephrolithiasis-induced renal fibrosis?

To investigate fibrotic lesions in renal tissues obtained from patients with large calculi, and to selectively evaluate the expression and clinical significance of Twist and E-cadherin in nephrolithiasis patients. We recruited 50 patients with kidney stone and 32 matched healthy controls. We determined plasma creatinine (Cr) and corrected Cr clearance (CCr). For the 50 patients, we detected daily urine protein excretion. At the end of percutaneous nephroscopic lithotomy, we performed puncture biopsy to acquire kidney tissue. We obtained normal control kidney tissues from non-nephrolithiasis patients who received a surgical biopsy during open surgery. We determined the expression of Twist and E-cadherin by immunohistochemical staining and scored it with clinical parameters. In addition, we analyzed the degree of expression of Twist and its correlation with long-term renal survival. Overall, the renal function of patients significantly decreased, as indicated by Cr and reduced CCr compared with healthy controls. Activated Twist was strongly expressed in tubular epithelial cells from kidneys of nephrolithiasis patients, whereas we found little positive staining of Twist in normal kidneys. Meanwhile, the expression of E-cadherin was significantly suppressed in kidneys of nephrolithiasis patients. Twist expression was inversely correlated with E-cadherin expression; using multivariate analysis, data showed that the factors influencing renal survival in patients were CCr (relative ratio, 4.39; 95% confidence interval, 1.34-14.38; P = 0.013) and the extent of Twist expression (relative ratio, 3.45; 95% confidence interval, 1.10-10.68; P = 0.033).

Our data suggest that the possible novel EMT marker molecule Twist and Twist staining might be a valuable index predicting renal fibrosis progression in human nephrolithiasis.