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Does the 10 Hounsfield units unenhanced computed tomography attenuation threshold apply to cortisol secreting adrenocortical adenomas? Computed tomography (CT) unenhanced attenuation value of <10 Hounsfield units (HU) has an excellent specificity (98%) to diagnose lipid-rich adrenocortical adenomas (ACAs) with a weaker sensitivity (71%). To determine from a routine clinical perspective if unenhanced attenuation value is influenced by cortisol secretion in ACAs. This was a retrospective study of cases collected between 2009 and 2012. This study was conducted in a tertiary-care university hospital. Seventy-two patients operated on for an ACA (Weiss score ≤ 2) were analysed. Thirty-four patients had an ACA oversecreting cortisol (Cush-ACA). Thirty-eight patients had an ACA without cortisol oversecretion (Non Hyper-ACA). CT unenhanced attenuation value was correlated with the functional status. The Weiss score items were analysed. Among the 34 patients with a Cush-ACA a minority (n = 7) had an unenhanced attenuation value under 10 HU. Among the high precontrast density (> 10 HU) Cush-ACAs, washout analysis after contrast administration was consistent with the benign nature of the tumor in ∼ 60% of the cases. Less than 25% clear cells (lipid-rich cells), a Weiss score item, was present in 50% of the Cush-ACAs in favour of a lipid-poor content.

Unenhanced attenuation value has a poor sensitivity to diagnose an ACA in case of cortisol oversecretion due to poor lipid content. Nevertheless, the accuracy of washout analysis was preserved in the group of Cush-ACAs.

Do patterns and predictors of urodynamics use in the United States? Due to the paucity of data on urodynamics on the national level, we assessed the use of urodynamics in a large sample of individuals in the United States and identified predictors of increased complexity of urodynamic procedures. Using administrative health care claims for adults enrolled in private insurance plans in the United States from 2002 to 2007, we identified those who underwent cystometrogram and abstracted relevant demographic and clinical data. We used logistic regression to identify predictors of higher urodynamic complexity over basic cystometrogram, specifically cystometrogram plus pressure flow study and videourodynamics. We identified 16,574 urodynamic studies, of which 23% were cystometrograms, 71% were cystometrograms plus pressure flow studies and 6% were videourodynamics. Stress incontinence was the most common clinical condition for all studies (33.7%), cystometrogram (30.8%), cystometrogram plus pressure flow study (35.4%) and videourodynamics (24.4%). Urologists performed 59.8% of all urodynamics and gynecologists performed 35.5%. Providers with 14 or more urodynamic studies during the study period performed 75% of all urodynamics and were more likely to perform cystometrogram plus pressure flow study and videourodynamics. On regression analysis the most consistent predictors of cystometrogram plus pressure flow study and/or videourodynamics over cystometrogram were specialty (urologist) and the number of urodynamic tests performed by the provider.

Most urodynamics in this series consisted of cystometrogram plus pressure flow study with stress incontinence the most common diagnosis. However, regardless of diagnosis, urologists and providers who performed more urodynamics were more likely to perform pressure flow study and/or videourodynamics in addition to cystometrogram. Further research is needed to determine whether these differences reflect gaps in the consistency or appropriateness of using urodynamics.

Is tRPV1 activation required for hypertonicity-stimulated inflammatory cytokine release in human corneal epithelial cells? To determine whether hypertonic stress promotes increases in inflammatory cytokine release through transient receptor potential vanilloid channel type 1 (TRPV1) signaling pathway activation in human corneal epithelial cells (HCECs). Hyperosmotic medium was prepared by supplementing isotonic Ringers solution with sucrose. Ca2+ signaling was measured in fura2-AM-loaded HCECs using a single-cell fluorescence imaging system. Western blot analysis evaluated the phosphorylation status of EGFR, ERK, p38 MAPK, and nuclear factor (NF)-κB. ELISA assessed the effect of TRPV1 activation on the release of IL-6 and IL-8. A 450 mOsm hypertonic stress elicited 2-fold Ca2+ transients that were suppressed by the TRPV1-selective antagonists capsazepine and JYL 1421. Such transients were enhanced by PGE2. Hypertonicity-induced EGF receptor (EGFR) transactivation was suppressed by preincubating HCECs with capsazepine, matrix metalloproteinase 1 (MMP1) inhibitor TIMP-1, broad-spectrum MMP inhibitor GM 6001, heparin-bound (HB)-EGF inhibitor CRM 197, or EGFR inhibitor AG 1478. ERK and p38 MAPK and NF-κB activation after EGFR transactivation occurred in tonicity and in a time-dependent manner. Hypertonicity-induced increases in IL-6 and IL-8 releases were suppressed by exposure to capsazepine, AG 1478, ERK inhibitor PD 98059, p38 inhibitor SB 203580, or NF-κB inhibitor PDTC.

Hypertonic stress-elicited TRPV1 channel stimulation mediates increases in a proinflammatory cytokine IL-6 and a chemoattractant IL-8 by eliciting EGFR transactivation, MAPK, and NF-κB activation. Selective drug modulation of either TRPV1 activity or its signaling mediators may yield a novel approach to suppressing inflammatory responses occurring in dry eye syndrome.

Is preoperative radiotherapy associated with worse functional results after coloanal anastomosis for rectal cancer? This study was designed to evaluate functional outcome in patients treated with preoperative radiotherapy after low anterior resection and a coloanal anastomosis for low rectal cancer. Functional outcome data from patients enrolled in a prospective randomized trial comparing 3 reconstructive procedures were evaluated with respect to administration of preoperative radiotherapy. Incontinence was assessed with a questionnaire on bowel function including the Fecal Incontinence Severity Index; sexual function was assessed with the Sexual Health Inventory for Men and a gender-specific questionnaire for women. Quality of life was assessed with SF-36 scores. Of 364 patients enrolled, 153 (42%) had no radiotherapy or chemotherapy, and 211 (58%) had preoperative radiotherapy; 186 (51%) had chemotherapy in addition to radiotherapy. Comparison of irradiated vs. nonirradiated patients showed no significant differences in postoperative morbidity (29.9% vs. 35.3%; P = 0.27). Two-year follow-up of 297 patients showed greater impairment of bowel function in irradiated patients (n = 170) vs. nonirradiated patients (n = 127): e.g., mean number of daily bowel movements at 12 months, 4.2 +/- 3.5 vs. 3.5 +/- 2.6, P = 0.032; urgency, 85% vs. 67%, P = 0.002). Antidiarrheal use was significantly higher in irradiated patients vs. nonirradiated patients at 4 (P = 0.043), 12 (P = 0.002), and 24 (P = 0.001) months. Sexual Health Inventory for Men scores indicated poorer function in irradiated patients at 24 months (P = 0.039). Preoperative radiotherapy had no deleterious effects on quality of life. Multivariate analyses showed that negative effects of preoperative radiotherapy on urgency at 4 months (P = 0.002) and antidiarrheal use at 24 months were independent of reconstruction technique, but a positive effect of reconstruction with a J-pouch was still observed in patients who received radiotherapy.

Preoperative radiotherapy does not increase overall morbidity but is associated with poorer functional outcome after low anterior resection with coloanal anastomosis. Preoperative radiotherapy and the J-pouch are nonconfounding predictors of functional outcome up to 24 months after surgery.

Does clinical utility of 3-dimensional echocardiography in the evaluation of tricuspid regurgitation caused by pacemaker lead? This study evaluated the usefulness of 3-dimensional echocardiography (3-DE) for identifying permanent pacemaker (PPM) or implantable cardioverter defibrillator (ICD) lead-related symptomatic tricuspid regurgitation (TR). Eighty-seven patients underwent 3-DE examination: 50 patients with PPM, 17 with ICD, and 20 with cardiac resynchronization therapy devices. TR severity was classified as trivial/mild, moderate, or severe according to the ratio of TR area to right atrium area. The 3-DE identified the lead route and position at the tricuspid valve in 82 patients (94.2%). In 5 patients, images without lead-induced artifacts could not be obtained. TR severity was trivial/mild in 50 patients, moderate in 20 patients, and severe in 12 patients. In all patients with trivial/mild TR and all but 1 patient with moderate TR, leads were positioned on the annulus side between leaflets. Lead-induced obstruction to tricuspid valve closing was identified in 1 patient with moderate TR and in 7 of 12 patients with severe TR: 4 patients had septal leaflet obstruction, and 4 had posterior leaflet obstruction.

The 3-DE can identify the lead route and position at the tricuspid valve and lead-related severe TR, so may be a useful technique of diagnosing the cause of severe TR in patients with PPM or ICD.

Is brain-specific fatty acid-binding protein elevated in serum of patients with dementia-related diseases? There is a need for biomarkers in accessible matrices, such as blood, for the diagnosis of neurodegenerative diseases. The aim of this study was to measure the serum levels of brain-type fatty acid-binding protein (FABP) and heart-type FABP in patients with dementia-involving diseases. Brain- and heart-type FABP were measured in serum samples from patients with either Alzheimer's disease (AD) (n = 31), Parkinson's disease (PD, n = 43), or other cognitive disorders (OCD, n = 42) and in 52 healthy controls. The localization of brain- and heart-type FABP was determined in brain sections by immunohistochemistry. Brain-type FABP levels were elevated in serum of 29%, 35%, and 24% of the patients with AD, PD, and OCD, respectively, and in 2% of the healthy donors. Heart-type FABP serum levels were not different amongst the patient groups. Brain-type and heart-type FABP expression was observed in reactive astrocytes in brain sections of patients with AD.

In contrast to heart-type FABP, serum levels of brain-type FABP are elevated in a significant proportion of patients with various neurodegenerative diseases and can therefore have importance for defining subgroups of these patients.

Is ocular tremor in Parkinson 's disease due to head oscillation? We investigated the origin of a recently reported ocular microtremor in patients with Parkinson's disease (PD). Eye movements were recorded in 2 unselected patients with PD. Two recording techniques were used to control for artifacts: infrared video-oculography and infrared scleral reflection techniques. Head movements were also recorded with 2 different accelerometers. We recorded ocular oscillations in both patients (microtremor). Ocular tremor was accompanied by a recordable (but clinically nonvisible) head tremor of equal fundamental frequency and high coherence with both the eye oscillation and a recordable limb tremor. The eye movements were in the opposite direction to the head oscillation (ie, compensatory) and were suppressed by head restraint. There was no subjective oscillopsia, nor ocular tremor on fundoscopy.

The "ocular tremor" observed in patients with PD disease is a compensatory eye movement secondary to transmitted head tremor, in agreement with clinical wisdom that these patients do not report oscillopsia.

Is erythrocyte sodium-lithium countertransport activity inversely correlated to adiponectin , retinol binding protein 4 and body height? We have previously described that the sodium/lithium countertransport (SLC) in the erythrocyte cell membrane is closely linked to obesity and insulin resistance. Adiponectin and retinol-binding protein 4 (RBP-4) are believed to affect obesity and insulin resistance. In the present study, we aimed to further characterize the relationship between SLC, inflammatory markers, adiponectin and RBP-4. We included 93 clinically healthy 58-year-old men selected to display variations in insulin sensitivity. High sensitivity C-reactive protein (hs-CRP), TNF-alpha, soluble TNF-alpha-receptors (sTNFR) 1 and 2, IL-6 and RBP-4 were measured using antibody-based techniques. Adiponectin was determined by a radioimmunoassay kit. The lithium concentration in the special flux medium was measured by atomic absorption spectrophotometry. In univariate analyses, SLC correlated negatively with RBP-4 (r(s) = -0.256, p = -0.017) and to adiponectin (r(s) = -0.316, p = 0.003) and positively with TNF-alpha (r(s) = 0.346, p = 0.001) and hs-CRP (r(s) = 0.288, p = 0.005). There were no statistically significant correlations with sTNFR 1 or 2 or IL-6. SLC was negatively associated to body height (r(s) = -0.256, p = 0.013).

We are the first to report that SLC correlates negatively with adiponectin and RBP-4. This finding is intriguing, as adiponectin is anti-inflammatory and anti-diabetic whereas RBP-4 supposedly decreases insulin sensitivity. We also observed a negative association between SLC activity and body height indicating that SLC activity is not primarily influenced by fat mass. The positive association of SLC with markers of inflammatory activity such as TNF-alpha and hs-CRP is in line with the proposed link between inflammation and insulin resistance.

Is [ Serum calgranulin C a highly sensitive autoinflammation activity indicator in patients with familial periodic fevers ]? To determine the possibility of using the serum proinflammatory calcium-binding protein, or calgranulin C (S100A12), to assess activity and therapeutic efficiency in patients with periodic disease (PD) and other familial periodic fevers (FPFs). Thirty-five patients with PD and other FPDs, which were verified by molecular genetic study, were examined. In accordance with the disease activity, the patients were divided into 2 groups. The investigators determined the concentration of S100A12 by solid-phase enzyme immunoassay and that of other acute-phase inflammatory markers (erythrocyte sedimentation rate (ERT), neutrophil counts, and fibrinogen and C-reactive protein (CRP) concentrations). The serum concentration of S100A12 in the stage of disease activity was 466.7 (265.22--851.7) ng/ml, which was significantly higher than in remission (244.29 (118.93--409.85) ng/ml (p=0.000002). The highest S100A12 concentrations were noted in the patients with PD; these were 758.95 (434.80--1035.95) ng/ml; the S100A12 level in the majority of PD patients even during remission remained moderately higher. An investigation of the relationship of A100A12 to genetic variants found no differences between the patients homozygous for M694V and those with other genotypes (p=0.37). Estimation of the time course of therapy-induced changes in the serum S100A12 concentration revealed its considerable reduction (р=0.0018). However, normalization of S100A12 levels was not achieved in PD. The remaining increased S100A12 concentration in these patients may be suggestive of the activity of PD despite the absence of its clinical manifestations. S100A12 as a highly sensitive marker allows more exact evaluation of the anti-inflammatory effect of therapy. The S100A12 identification of the subclinical activity of autoinflammatory diseases made all the more important since traditional inflammatory markers, such as ERT, CRP, fibrinogen, and leukocyte counts, are less sensitive for these purposes. In our study, these markers were within the reference range in remission. No differences were found in the S100A12 levels between the groups with and without amyloidosis (p=0.62).

S100A12 is a highly sensitive marker for the activity of autoinflammatory diseases and the efficiency of their therapy. The serum level of S100A12 in PD may be used to diagnose the subclinical activity of inflammation, which is of importance in monitoring the risk of amyloidosis.

Are crime and perceptions of safety in the home neighborhood independently associated with physical activity among 11-15 year olds? To determine whether perceptions of neighborhood safety and measures of neighborhood crime are independently associated within physical activity in youth. The study sample consisted of 14,125 youths in grades 6-10 (ages 11-15) who participated in the nationally representative cross-sectional 2009/10 Canadian Health Behaviour in School-Aged Children Survey. Participants responded to four questions about how safe their home neighborhood is and reported whether or not they accumulated at least 4h/week of physical activity in their free-time outside of school hours. Crimes against persons was assessed in 1 km radius buffers around participants' homes. Associations were assessed using logistic regression. After controlling for crime and relevant confounders, the relative odds of being physically active outside of school was 0.52 (95% CI: 0.44-0.62) in youth whose perceptions of neighborhood safety were in the lowest quintile. After controlling for perceptions of safety and relevant confounders, the relative odds of being physically active outside of school was 0.75 (0.60-0.95) in youth from neighborhoods with crimes against persons scores in the highest quintile.

Within this large sample of 11-15 year olds, perceptions and objective measures of neighborhood safety and crime were independently associated with physical activity in free-time outside of school.

Does an analogue of piperonyl butoxide facilitate the characterisation of metabolic resistance? Previous work has demonstrated that piperonyl butoxide (PBO) not only inhibits microsomal oxidases but also resistance-associated esterases. The ability to inhibit both major metabolic resistance enzymes makes it an ideal synergist to enhance xenobiotics but negates the ability to differentiate which enzyme group is responsible for conferring resistance. This study examines an analogue that retains the ability to inhibit esterases but is restricted in its ability to act on microsomal oxidases, thus allowing an informed decision on resistance enzymes to be made when used in conjunction with the parent molecule.

Using examples of resistant insects with well-characterised resistance mechanisms, a combination of PBO and analogue allows identification of the metabolic mechanism responsible for conferring resistance. The relative potency of PBO as both an esterase inhibitor and an oxidase inhibitor is also discussed.

Does positive antibody response to vaccination in adolescence predict lower C-reactive protein concentration in young adulthood in the Philippines? Inflammation has been associated with a wide range of chronic degenerative diseases, but the developmental factors contributing to the regulation of inflammation are poorly understood. This study investigates the within-individual association between antibody response to vaccination in adolescence and C-reactive protein (CRP) concentration in young adulthood. In 1998-99, at age 14-15 years, a subset of participants (N = 96) in the Cebu Longitudinal Health and Nutrition Survey were administered a typhoid vaccine, and baseline and follow-up blood samples were drawn to assess the strength of the antibody response to vaccination. In 2005, at age 20-21 years, blood samples were drawn from the full cohort for measurement of CRP. N = 74 individuals had complete data at both time points. Bivariate associations and multivariate logistic regression models were evaluated to test the hypothesis that vaccine responsiveness in adolescence was significantly associated with CRP level in young adulthood. There was a strong and statistically significant association between antibody response to vaccination in adolescence and CRP in young adulthood. Median CRP was more than four times higher among nonresponders than responders, and nonresponders were 2.3 to 3.6 times more likely to have CRP in the top tertile of the sample distribution.

This study provides evidence for a prospective, within-individual link between more effective antibody-mediated immune defenses and lower levels of inflammation. In the context of prior research in this population, these results suggest that early environments are important determinants of multiple aspects of an individual's immuno-phenotype.

Do phosphoinositide 3-kinases p110α and p110β have differential roles in insulin-like growth factor-1-mediated Akt phosphorylation and platelet priming? Platelet hyperactivity is a contributing factor in the pathogenesis of cardiovascular disease and can be induced by elevated levels of circulating growth factors, such as insulin-like growth factor-1 (IGF-1). IGF-1 is a primer that cannot stimulate platelet activation by itself, but in combination with physiological stimuli can potentiate platelet functional responses via a phosphoinositide 3-kinase-dependent mechanism. In this study, we explored the role of the phosphoinositide 3-kinase p110α isoform in IGF-1-mediated enhancement of platelet function. Using a platelet-specific p110α knockout murine model, we demonstrate that genetic deletion, similar to pharmacological inactivation of p110α, did not affect proteinase-activated receptor 4 signaling to Akt/protein kinase B but significantly reduced IGF-1-mediated Akt phosphorylation. The p110β inhibitor TGX-221 abolished IGF-1-induced Akt phosphorylation in p110α-deficient platelets, demonstrating that both p110α and p110β contribute to IGF-1-mediated Akt phosphorylation. Genetic deletion of p110α had no effect on IGF-1-mediated increases in thrombus formation on collagen and enhancement of proteinase-activated receptor 4-mediated integrin activation and α-granule secretion. In contrast, pharmacological inhibition of p110α blocked IGF-1-mediated potentiation of integrin activation and α-granule secretion. Functional enhancement by IGF-1 in p110α knockout samples was lost after TGX-221 treatment, suggesting that p110β drives priming in the absence of the p110α isoform.

Together, these results demonstrate that both p110α and p110β are involved in Akt signaling by IGF-1, but that it is the p110α isoform that is responsible for IGF-1-mediated potentiation of platelet function.

Do biodegradable microparticles containing crotamine isolated from Crotalus durissus terrificus display antileishmanial activity in vitro? To evaluate antileishmanial activity of crotamine, a toxin isolated from Crotalus durissus terrificus, in solution form and encapsulated in biodegradable microparticles in vitro. Particles were analyzed on-chip by surface plasmon resonance and characterized by testing their diameters, zeta potential and encapsulation rate. The viability of promastigotes as well as murine macrophages was assessed. Furthermore, the phagocytic index was determined for macrophages, and cell supernatants were collected for the determination of TNF-α levels. An infection assay using Leishmania amazonensis-infected macrophages was also conducted. The diameters and zeta potential of control particles (1.35 μm; -12.3 mV) and of those containing crotamine (3.09 μm; -20.9 mV) were adequate for the assays conducted. Crotamine-loaded particles were better captured by macrophages than control particles (increase of 12% in the phagocytic index), leading to increased TNF-α levels (196 pg/ml), and they also induced a significant decrease in the numbers of amastigotes compared to infected macrophages only.

The approach presented here opens the possibility of working with safe concentrations of encapsulated toxins to reach antileishmanial effects.

Does anterior Shoulder Instability be Associated With an Underlying Deficiency of the Bony Glenoid Concavity? To determine whether anterior shoulder instability is associated with an inherent deficiency of the bony glenoid concavity, which results in a reduced bony shoulder stability ratio (BSSR). In this case-control study, we searched the institutional database for patients treated for unilateral recurrent anterior shoulder instability. We included 30 consecutive patients with atraumatic instability, 30 consecutive patients with traumatic instability, and 36 matched healthy controls, for a total of 96 shoulders. Computed tomography images of the unaffected shoulders of the instability patients were compared with images of the ipsilateral shoulders of age- and sex-matched healthy controls for differences in glenoid morphology. By use of a mathematical formula based on Pythagorean trigonometric identities, the mean BSSRs of the different groups were calculated and compared. Validation of the formula was accomplished by finite element analysis. The mean BSSR of atraumatic instability patients was 17.9% ± 8.5% and therefore significantly lower than the mean BSSR of 31.1% ± 7.5% of the control group (13.2%; 95% confidence interval [CI], 9.1% to 17.4%; P < .001). The mean BSSR of the traumatic instability group was higher, at 23.9% ± 8.5% (P = .007), but still showed a deficit of 7.2% (95% CI, 2.8% to 11.7%; P = .002) compared with controls. The atraumatic instability group showed a mean reduction of 0.9 mm (95% CI, 0.6 to 1.1 mm; P < .001) in concavity depth and a decrease of 2.9° (95% CI, 0.4° to 5.3°; P = .021) in concavity retroversion, whereas the traumatic instability patients had a reduction of 0.4 mm (95% CI, 0.1 to 0.8 mm; P = .006) in concavity depth. Neither of the instability groups differed significantly from their respective controls in terms of glenoid concavity diameter, head radius, or glenoid vault morphology.

Anterior shoulder instability is associated with an inherent flattening of the bony glenoid concavity, which significantly decreases the BSSR. The deficiency appears to be more pronounced in patients with atraumatic instability than in patients with traumatic instability.

Do methyltransferases acquired by lactococcal 936-type phage provide protection against restriction endonuclease activity? So-called 936-type phages are among the most frequently isolated phages in dairy facilities utilising Lactococcus lactis starter cultures. Despite extensive efforts to control phage proliferation and decades of research, these phages continue to negatively impact cheese production in terms of the final product quality and consequently, monetary return. Whole genome sequencing and in silico analysis of three 936-type phage genomes identified several putative (orphan) methyltransferase (MTase)-encoding genes located within the packaging and replication regions of the genome. Utilising SMRT sequencing, methylome analysis was performed on all three phages, allowing the identification of adenine modifications consistent with N-6 methyladenine sequence methylation, which in some cases could be attributed to these phage-encoded MTases. Heterologous gene expression revealed that M.Phi145I/M.Phi93I and M.Phi93DAM, encoded by genes located within the packaging module, provide protection against the restriction enzymes HphI and DpnII, respectively, representing the first functional MTases identified in members of 936-type phages.

SMRT sequencing technology enabled the identification of the target motifs of MTases encoded by the genomes of three lytic 936-type phages and these MTases represent the first functional MTases identified in this species of phage. The presence of these MTase-encoding genes on 936-type phage genomes is assumed to represent an adaptive response to circumvent host encoded restriction-modification systems thereby increasing the fitness of the phages in a dynamic dairy environment.

Does fibroblast growth factor receptor-1 signaling induce osteopontin expression and vascular smooth muscle cell-dependent adventitial fibroblast migration in vitro? Increased expression of osteopontin (OPN), fibroblast growth factors (FGFs), and their type-1 receptor (FGFR-1) is associated with neointima formation and atherosclerosis. This study tested the hypothesis that ligand activation of FGFR-1 stimulates OPN expression in rat aortic smooth muscle cells (RASMCs), explored the signaling pathway involved, and assessed the functional consequences of activating this pathway on adventitial fibroblast (AF) migration in vitro. Exogenous FGF-1 stimulated expression of OPN mRNA and protein in RASMCs in vitro in a dose- and time-dependent manner. OPN mRNA induction by FGF-1 was completely inhibited by either actinomycin D or cycloheximide, selective inhibitors of RNA polymerase and protein synthesis, respectively. OPN mRNA induction by FGF-1 was attenuated by PD 166866, a highly selective and potent FGFR-1 tyrosine kinase inhibitor. Addition of either PP2 or PD98059, specific inhibitors of Src and mitogen-activated extracellular signal-regulated kinase (MEK)/mitogen-activated protein (MAP) kinases, respectively, attenuated FGF-1-stimulated OPN mRNA expression. FGF-1 treatment of RASMCs enhanced RASMC-conditioned medium-stimulated AF migration; this effect was inhibited by pretreatment of RASMCs with either PD166866 or PP2. Immunodepletion of OPN from RASMC-conditioned medium inhibited both basal and FGF-1-stimulated AF migration.

This in vitro study provided a first indication that ligand-activated FGFR-1 plays a significant role in upregulation of OPN expression at the transcriptional level via signaling to Src/MEK/MAP kinases in RASMCs and that this pathway is functionally significant in mediating AF migration via stimulation of OPN expression.

Does nucleotide oligomerization domain 2 contribute to the innate immune response in THCE cells stimulated by Aspergillus fumigatus conidia? To investigate the expression of nucleotide oligomerization domain 2 (NOD2) in the immortalized human corneal epithelial cell line (THCE), and its role in the innate immune response triggered by inactive Aspergillus fumigatus (Af) conidia. The normal THCE cells were investigated as controls. After incubation with inactive Af conidia for 0.5, 2, 4, 6, and 8 hours, THCE cells were harvested, mRNA expression of NOD2 and receptor interacting protein 2 (RIP2) was detected by RT-PCR. Intracellular proteins including NOD2, NF-κB and proinflammatory cytokines such as TNF-α, IL-8, IL-6 in the cell supernatant were analyzed by ELISA. Our data indicate that NOD2 expressed in the normal THCE cells. After triggered by the inactive Af conidia, the expression of NOD2, RIP2 mRNA and the secretion of NOD2, NF-κB, TNF-α, IL-8, IL-6 both increased in a time-depended manner, and reached the peak point at 4, 6, 6, 4, 6, 6, 4 hours, respectively. And after pretreated with NOD2 neutralizing antibody, the expression of RIP2, NF-κB, TNF-α, IL-8 both decreased dramatically at the peak point, while the secretion of IL-6 changed little.

The results of this study suggest that NOD2 exists and expresses in the THCE cells, and contributes to the innate immune responses triggered by inactive Af conidia by induction of proinflammatory cytokines such as TNF-α and IL-8 through the NF-κB pathway.

Does thrombin induce heme oxygenase-1 expression in human synovial fibroblasts through protease-activated receptor signaling pathways? Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, and proinflammatory processes. Abnormalities in these processes are primary features of osteoarthritis (OA). Heme oxygenase (HO)-1 is a stress-inducible rate-limiting enzyme in heme degradation that confers cytoprotection against oxidative injury. Here, we investigated the intracellular signaling pathways involved in thrombin-induced HO-1 expression in human synovial fibroblasts (SFs). Thrombin-mediated HO-1 expression was assessed with quantitative real-time (q)PCR. The mechanisms of action of thrombin in different signaling pathways were studied by using Western blotting. Knockdown of protease-activated receptor (PAR) proteins was achieved by transfection with siRNA. Chromatin immunoprecipitation assays were used to study in vivo binding of Nrf2 to the HO-1 promoter. Transient transfection was used to examine HO-1 activity. Osteoarthritis synovial fibroblasts (OASFs) showed significant expression of thrombin, and expression was higher than in normal SFs. OASFs stimulation with thrombin induced concentration- and time-dependent increases in HO-1 expression. Pharmacologic inhibitors or activators and genetic inhibition by siRNA of protease-activated receptors (PARs) revealed that the PAR1 and PAR3 receptors, but not the PAR4 receptor, are involved in thrombin-mediated upregulation of HO-1. Thrombin-mediated HO-1 expression was attenuated by thrombin inhibitor (PPACK), PKCδ inhibitor (rottlerin), or c-Src inhibitor (PP2). Stimulation of cells with thrombin increased PKCδ, c-Src, and Nrf2 activation.

Our results suggest that the interaction between thrombin and PAR1/PAR3 increases HO-1 expression in human synovial fibroblasts through the PKCδ, c-Src, and Nrf2 signaling pathways.

Do transient receptor potential canonical type 3 channels facilitate endothelium-derived hyperpolarization-mediated resistance artery vasodilator activity? Microdomain signalling mechanisms underlie key aspects of artery function and the modulation of intracellular calcium, with transient receptor potential (TRP) channels playing an integral role. This study determines the distribution and role of TRP canonical type 3 (C3) channels in the control of endothelium-derived hyperpolarization (EDH)-mediated vasodilator tone in rat mesenteric artery. TRPC3 antibody specificity was verified using rat tissue, human embryonic kidney (HEK)-293 cells stably transfected with mouse TRPC3 cDNA, and TRPC3 knock-out (KO) mouse tissue using western blotting and confocal and ultrastructural immunohistochemistry. TRPC3-Pyr3 (ethyl-1-(4-(2,3,3-trichloroacrylamide)phenyl)-5-(trifluoromethyl)-1H-pyrazole-4-carboxylate) specificity was verified using patch clamp of mouse mesenteric artery endothelial and TRPC3-transfected HEK cells, and TRPC3 KO and wild-type mouse aortic endothelial cell calcium imaging and mesenteric artery pressure myography. TRPC3 distribution, expression, and role in EDH-mediated function were examined in rat mesenteric artery using immunohistochemistry and western blotting, and pressure myography and endothelial cell membrane potential recordings. In rat mesenteric artery, TRPC3 was diffusely distributed in the endothelium, with approximately five-fold higher expression at potential myoendothelial microdomain contact sites, and immunoelectron microscopy confirmed TRPC3 at these sites. Western blotting and endothelial damage confirmed primary endothelial TRPC3 expression. In rat mesenteric artery endothelial cells, Pyr3 inhibited hyperpolarization generation, and with individual SK(Ca) (apamin) or IK(Ca) (TRAM-34) block, Pyr3 abolished the residual respective IK(Ca)- and SK(Ca)-dependent EDH-mediated vasodilation.

The spatial localization of TRPC3 and associated channels, receptors, and calcium stores are integral for myoendothelial microdomain function. TRPC3 facilitates endothelial SK(Ca) and IK(Ca) activation, as key components of EDH-mediated vasodilator activity and for regulating mesenteric artery tone.

Is renal dysfunction common following resuscitation from out-of-hospital cardiac arrest? Cardiac arrest patients often suffer from dysfunction of multiple organ systems after return of spontaneous circulation (ROSC). The incidence of renal dysfunction in patients with out-of-hospital cardiac arrest (OHCA) is not well described. Renal dysfunction has been associated with worse outcomes in critical illness. Renal dysfunction is common after OHCA, and renal dysfunction is independently associated with survival. We performed a retrospective review of consecutive adult patients admitted to an intensive care unit after successful resuscitation from OHCA between 01/01/2005 and 12/31/2010. Patients were excluded for death or withdrawal of care within 24h, preexisting end-stage renal disease, or OHCA due to hyperkalemia. The RIFLE criteria were used to classify subjects with renal dysfunction into one of three categories - risk, injury, or failure - based on trending of serum creatinine concentration or glomerular filtration rate. Data were analyzed using descriptive statistics. Of 364 patients, 38 were excluded due to death or withdrawal of care within 24h, 11 due to end-stage renal disease, and 4 due to OHCA from hyperkalemia, leaving 311 patients in the final analysis. The mean age was 58 (SD 16) years; 174 (59%) were male, VF/VT was the primary rhythm in 156 (50%), and 236 (80%) were comatose at hospital arrival. Among 311 patients, 32 (10.3%) developed acute renal failure (ARF), 27 (8.7%) developed acute kidney injury (AKI), and 56 (18.0%) developed risk of renal dysfunction. Of the 32 subjects that developed ARF, renal replacement therapy (RRT) was initiated on 13 (40.6%). Development of ARF was not associated with survival (OR 0.73; 95% CI 0.26, 2.05) after adjusting for initial rhythm or illness severity.

More than one-third of patients resuscitated from OHCA developed evidence of renal dysfunction, and 19% of patients meeting criteria for AKI or ARF. In this study, development of renal failure was not independently associated with survival.

Are serum mannose-binding lectin levels decreased in behcet 's disease and associated with disease severity? To investigate serum levels of mannose-binding lectin (MBL), a complement-like protein of collectin family, in patients with Behcet's disease (BD). MBL levels were measured in sera of 130 patients with BD, 64 patients with recurrent oral ulcerations (ROU), and 105 healthy controls (HC) with ELISA. Patients with BD had significantly lower median serum MBL levels compared to HC (1857 vs 3136 ng/ml, p = 0.001). No significant difference was observed in median serum MBL levels between BD and ROU (2309 ng/ml, p = 0.252). Low MBL levels (</= 500 ng/ml) were present in a higher proportion of BD patients compared to HC (29% vs 16%, p = 0.021). A severe disease course (total clinical severity score >/= 4) was more frequently observed in BD patients with very low serum MBL levels (</= 100 ng/ml) (19% vs 6%, p = 0.046). When serum MBL levels were analyzed separately according to gender, the frequency of vascular disease was higher in men with very low serum MBL levels (80% vs 42%, p = 0.042).

MBL deficiency might contribute to the pathogenesis of BD and affect its clinical course.

Does total parenteral nutrition supplementation with glutamine improve survival after gut ischemia/reperfusion? Total parenteral nutrition (TPN) alters gut cytokines and mucosal immunity and increases intercellular adhesion molecule-1 (ICAM-1) expression, gut neutrophil levels, and mortality after gut ischemia. Supplementation of TPN with glutamine partially supports mucosal immunity by preserving respiratory and intestinal IgA levels, maintaining the proper IgA-stimulating cytokine milieu within the intestine, and reducing intestinal ICAM-1 expression and neutrophil accumulation. This work investigates whether glutamine supplementation of TPN affects mortality in mice after gut ischemic insult. Thirty-eight mice were randomized to receive chow, TPN, or 2% glutamine-supplemented TPN (GLN-TPN) for 5 days. After feeding their respective diets, gut ischemia/reperfusion (I/R) was induced with superior mesenteric artery occlusion for 30 minutes followed by resuscitation with 1 mL saline. Survival was recorded until 72 hours after reperfusion. Survival time was significantly reduced in the TPN-fed mice compared with both chow-fed and GLN-TPN-fed mice (p < .05). Survival at 72 hours after reperfusion was also significantly lower in the TPN-fed mice than in the chow-fed and GLN-TPN-fed mice (p < .05)

Glutamine supplementation of TPN significantly improves survival after gut I/R, suggesting modulation of the inflammatory response or improved gut tolerance to low-flow states.

Is a polymorphism located at an ATG transcription start site of the heme oxygenase-2 gene associated with classical Parkinson 's disease? Oxidative stress and iron deposition is related to Parkinson's disease (PD). Heme oxygenase 2 (HMOX2) catalyzes the cleavage of the heme ring to form biliverdin with release of iron and carbon monoxide. This study aims to analyze variations in the HMOX2 gene in patients with PD. We mapped four single nucleotide polymorphisms (SNPs) and copy number variations of the HMOX2 gene in 691 patients with PD and 747 healthy individuals. We identified a highly homogeneous association of the HMOX2 SNP rs2270363 homozygous G/G genotype with patients with classical PD phenotype compared with healthy individuals. We identified three patients with PD and two control individuals with a single copy of the HMOX2 gene. No individuals with zero or more than two gene copies were identified.

We describe for the first time, copy number variations in the HMOX2 gene and an association of the SNP rs2270363 with PD risk.

Does fAK regulate intestinal epithelial cell survival and proliferation during mucosal wound healing? Following damage to the intestinal epithelium, restoration of epithelial barrier integrity is triggered by a robust proliferative response. In other tissues, focal adhesion kinase (FAK) regulates many of the cellular processes that are critical for epithelial homeostasis and restitution, including cell migration, proliferation and survival. However, few studies to date have determined how FAK contributes to mucosal wound healing in vivo. To examine the role of FAK in intestinal epithelial homeostasis and during injury, we generated intestinal epithelium (IE)-specific conditional FAK knockout mice. Colitis was induced with dextran-sulfate-sodium (DSS) and intestinal tissues were analyzed by immunohistochemistry and immunoblotting. While intestinal development occurred normally in mice lacking FAK, FAK-deficient animals were profoundly susceptible to colitis. The loss of epithelial FAK resulted in elevated p53 expression and an increased sensitivity to apoptosis, coincident with a failure to upregulate epithelial cell proliferation. FAK has been reported to function as a mechanosensor, inducing cyclin D1 expression and promoting cell cycle progression under conditions in which tissue/matrix stiffness is increased. Collagen deposition, a hallmark of inflammatory injury resulting in increased tissue rigidity, was observed in control and FAK knockout mice during colitis. Despite this fibrotic response, the colonic epithelium in FAK-deficient mice exhibited significantly reduced cyclin D1 expression, suggesting that proliferation is uncoupled from fibrosis in the absence of FAK. In support of this hypothesis, proliferation of Caco-2 cells increased proportionally with matrix stiffness in vitro only under conditions of normal FAK expression; FAK depleted cells exhibited reduced proliferation concomitant with attenuated cyclin D1 expression.

In the colon, FAK functions as a regulator of epithelial cell survival and proliferation under conditions of mucosal injury and a mechanosensor of tissue compliance, inducing repair-driven proliferation in the colonic epithelium through upregulation of cyclin D1.

Is separation of somatic and germ cells required to establish primate spermatogonial cultures? Can primate spermatogonial cultures be optimized by application of separation steps and well defined culture conditions?

We identified the cell fraction which provides the best source for primate spermatogonia when prolonged culture is desired.

Are metabolically protective cytokines adiponectin and fibroblast growth factor-21 increased by acute overfeeding in healthy humans? Circulating levels of metabolically protective and adverse cytokines are altered in obese humans and rodent models. However, it is not clear whether these cytokines are altered rapidly in response to over-nutrition, or as a later consequence of the obese state. Forty sedentary healthy individuals were examined prior to and at 3 and 28 days of high fat overfeeding (+1250 kCal/day, 45% fat). Insulin sensitivity (hyperinsulinaemic-euglycaemic clamp), adiposity, serum levels of adiponectin and fibroblast growth factor-21 (FGF21), fatty acid binding protein-4 (FABP4), lipocalin-2 and plasminogen activator factor-1 (PAI1) were assessed. Statistics were performed by repeated measures ANOVA. Overfeeding increased weight, body fat and liver fat, fasting glucose, insulin and reduced insulin sensitivity by clamp (all P <0.05). Metabolically protective cytokines, adiponectin and FGF21 were increased at day 3 of overfeeding (P ≤0.001) and adiponectin was also elevated at day 28 (P=0.001). FABP4, lipocalin-2 and PAI-1 were not changed by overfeeding at either time point.

Metabolically protective cytokines, adiponectin and FGF-21, were increased by over nutrition and weight gain in healthy humans, despite increases in insulin resistance. We speculate that this was in attempt to maintain glucose homeostasis in a state of nutritional excess. PAI-I, FABP4 and lipocalin 2 were not altered by overfeeding suggesting that changes in these cytokines may be a later consequence of the obese state.

Is maternal-fetal medicine specialist density inversely associated with maternal mortality ratios? Our study's objective was to determine the relationship between state-specific maternal mortality ratios and the density of maternal-fetal medicine specialists. State maternal mortality ratios from 1994 to 2001 were calculated from the Centers for Disease Control and Prevention WONDER database. Practitioner distribution data were obtained from professional associations. Demographic information regarding states was gathered from the 2000 US census data. Bivariable and multivariable analyses were conducted with the use of Spearman correlations and Poisson regression, respectively. The median state maternal-mortality ratio was 7.5/100,000 live births. Our study showed that an increase of 5 maternal-fetal specialists per 10,000 live births results in a 27% reduction in the risk of maternal death (relative risk [RR] = 0.73, 95% CI = 0.58-0.93, P = 0.012). This risk reduction was based on a multivariable Poisson regression model that included the following variables and their significant interactions: state-specific percentages of mothers in poverty, mothers without a high school diploma, minority mothers, and teenage mothers.

The density of maternal-fetal medicine specialists is significantly and inversely associated with maternal mortality ratios, even after controlling for state-level measures of maternal poverty, education, race, age, and their significant interactions.

Is vitamin D deficiency associated with an increased autoimmune response in healthy individuals and in patients with systemic lupus erythematosus? Vitamin D deficiency is widespread and has been associated with many chronic diseases, including autoimmune disorders. A study was undertaken to explore the impact of low vitamin D levels on autoantibody production in healthy individuals, as well as B cell hyperactivity and interferon α (IFNα) activity in patients with systemic lupus erythematosus (SLE). Serum samples from 32 European American female patients with SLE and 32 matched controls were tested for 25-hydroxyvitamin D (25(OH)D) levels, lupus-associated autoantibodies and serum IFNα activity. Isolated peripheral blood mononuclear cells were tested for intracellular phospho-ERK 1/2 as a measure of B cell activation status. Vitamin D deficiency (25(OH)D <20 ng/ml) was significantly more frequent among patients with SLE (n=32, 69%) and antinuclear antibody (ANA)-positive controls (n=14, 71%) compared with ANA-negative controls (n=18, 22%) (OR 7.7, 95% CI 2.0 to 29.4, p=0.003 and OR 8.8, 95% CI 1.8 to 43.6, p=0.011, respectively). Patients with high B cell activation had lower mean (SD) 25(OH)D levels than patients with low B cell activation (17.2 (5.1) vs 24.2 (3.9) ng/ml; p=0.009). Patients with vitamin D deficiency also had higher mean (SD) serum IFNα activity than patients without vitamin D deficiency (3.5 (6.6) vs 0.3 (0.3); p=0.02).

The observation that ANA-positive healthy controls are significantly more likely to be deficient in vitamin D than ANA-negative healthy controls, together with the finding that vitamin D deficiency is associated with certain immune abnormalities in SLE, suggests that vitamin D plays an important role in autoantibody production and SLE pathogenesis.

Is excessive lowering of blood pressure beneficial for progression of brain white matter hyperintensive and cognitive impairment in elderly hypertensive patients : 4-year follow-up study? This study was designed to explore the appropriate blood pressure (BP) target required to reduce cognitive decline and brain white matter lesions (WMLs) in elderly hypertensive patients. Elderly patients (n = 294, ≥80 years of age) being treated for hypertension were enrolled in a longitudinal study examining cognitive impairment after an initial assessment and a period of 4 years. All patients underwent neurological and cognitive assessment, laboratory examination, and magnetic resonance imaging of the brain. The 4-year follow-up examination revealed that body mass index, alcohol consumption, systolic blood pressure (SBP), diastolic blood pressure, and Mini-Mental State Examination (MMSE) all showed a significant decline, whereas fasting plasma glucose, white matter hyperintensities (WMH) volume, and the WMH/total intracranial volume (TIV) ratio were significantly increased when compared with baseline observations. Interestingly, the decline in MMSE, as well as the increment of WMH and WMH/TIV ratio was smaller in patients with SBP ranging from 140 to 160 mm Hg than in those whose SBP was lower than 140 mm Hg or higher than 160 mm Hg (P < .05). Furthermore, we observed that a 15 to 35 mm Hg targeted lowering of SBP in the elderly patients was more beneficial to our cognitive analysis than treatments that achieved less than 15 mm Hg or greater than 35 mm Hg (P < .05).

In elderly hypertensive patients, there exists a beneficial target for SBP lowering beyond which treatment may not be beneficial for improving or delaying the progression of cognitive impairment and WMLs.

Does qRS prolongation induced by cardiac resynchronization therapy correlate with deterioration in left ventricular function? The benefits of cardiac resynchronization in inducing reverse ventricular remodeling in patients with left ventricular (LV) systolic dysfunction have been well established. Still, up to 30% of the patients fail to derive significant improvement from this therapy. A subset of "nonresponders" experience deterioration in LV function following cardiac resynchronization therapy (CRT). Characteristics of this patient population, however, have not been studied. To determine characteristics of patients who experience deterioration in LV function following CRT. Clinical, electrocardiographic, and echocardiographic data were collected in 856 consecutive patients presenting for a new CRT device. For inclusion, all patients had an LV ejection fraction '40%, a QRS duration '120 ms, and available baseline and follow-up echocardiograms and electrocardiograms. Deterioration in LV function was defined as an absolute decrease of 5% or greater in ejection fraction from baseline. Multivariate models were constructed to identify variables significantly associated with deterioration. A total of 507 patients met inclusion criteria, of which 60 (11.8%) met criteria for deterioration. Patients with deterioration were more likely to be men (86.7% vs 66.9%; P = .002), have a non-left bundle branch block morphology (41.7% vs 23.7%; P = .001), and a history of atrial fibrillation (66.7% vs 51.7%; P = .03). On comparing the pre-CRT QRS duration with the first biventricular-paced QRS duration post-CRT implant, it was found that patients with LV deterioration had significant QRS widening than did those without deterioration (ms) (+3.9 ± 34.1 vs -9.0 ± 27.4, P = .007, respectively). In multivariate analysis, QRS widening indexed to the baseline QRS duration was significantly associated with LV deterioration (odds ratio 1.14 [1.06-1.23]; P = .001).

QRS widening is associated with deterioration in LV function following CRT.

Do [ Molecular characteristics and antibiotic resistance of Vibrio cholerae O139 in Shandong province ]? To investigate the molecular epidemiological characteristics and antibiotic resistance profiles of Vibrio cholerae O139 in Shandong province. A total of 13 strains of V. cholerae O139 (9 clinical strains and 4 environmental strains) isolated from cholera epidemics in Shandong province since 1997 were recovered and confirmed with serum agglutination and biochemical reaction. Pulsed-field gel electrophoresis (PFGE) was carried out for molecular subtyping. Virulence genes and drug resistance related genes were detected by PCR. Antibiotic susceptibility tests were performed using micro-broth dilution method. Thirteen strains of V. cholerae O139 were differentiated into seven pulsetypes. One clinical strain and two environmental strains isolated from Jining in 2013 were clustered into the pulsetype namely KZGN11O139. CN0077, and an identical PFGE pattern of KZGN11O139. CN0002 was found among three clinical strains from Jinan in 2005, Jining in 2005 and Heze in 2009. Other pulsotypes were unique in China and found only in Shandong province. Because of deletion of ctxAB and tcpI, the PFGE patterns of two strains isolated from Yantai in 2000 and 2004 were different from other 11 strains which harbored ctxAB, tcpA, tcpI, rtxA, hlyA and toxR. All strains contained one or more drug resistance related genes such as intI 1, intI 4 and sxt, and were resistant to two kinds of antibiotics at least. Among the 12 kinds of antibiotics, the resistant ratioes to kamamycin, trimethoprim-sulfamethoxazole, ampicillin and gentamicin were 11/13, 9/13, 7/13 and 7/13, respectively.

Molecular subtyping indicates possible epidemiological links among V.cholerae O139 in Shandong province, and almost all strains were toxigenic and drug resistant.

Are gastroesophageal reflux disease symptoms and dietary behaviors significant correlates of short sleep duration in the general population : the Nagahama Study? To examine relationships among gastroesophageal reflux disease (GERD) symptoms, dietary behaviors, and sleep duration in the general population. Cross-sectional. Community-based. There were 9,643 participants selected from the general population (54 ± 13 y). None. Sleep duration, sleep habits, and unfavorable dietary behaviors of each participant were assessed with a structured questionnaire. Participants were categorized into five groups according to their sleep duration: less than 5 h, 5 to less than 6 h, 6 to less than 7 h, 7 to less than 8 h, and 8 or more h per day. GERD was evaluated using the Frequency Scale for the Symptoms of GERD (FSSG) and participants having an FSSG score of 8 or more or those under treatment of GERD were defined as having GERD. Trend analysis showed that both the FSSG score and the number of unfavorable dietary habits increased with decreasing sleep duration. Further, multiple logistic regression analysis showed that both the presence of GERD (odds ratio = 1.19, 95% confidence interval (CI) = 1.07-1.32) and the number of unfavorable dietary behaviors (odds ratio = 1.19, 95% CI = 1.13-1.26) were independent and potent factors to identify participants with short sleep duration even after controlling for other confounding factors.

The current study showed that both GERD symptoms and unfavorable dietary behaviors were significant correlates of short sleep duration independently of each other in a large sample from the general population.

Do protease inhibitors enhance extracellular collagen fibril deposition in human mesenchymal stem cells? Collagen is a widely used naturally occurring biomaterial for scaffolding, whereas mesenchymal stem cells (MSCs) represent a promising cell source in tissue engineering and regenerative medicine. It is generally known that cells are able to remodel their environment by simultaneous degradation of the scaffolds and deposition of newly synthesized extracellular matrix. Nevertheless, the interactions between MSCs and collagen biomaterials are poorly known, and the strategies enhancing the extracellular matrix deposition are yet to be defined. In this study, we aim to investigate the fate of collagen when it is in contact with MSCs and hypothesize that protease inhibition will enhance their extracellular deposition of collagen fibrils. Specifically, human MSCs (hMSCs) were exposed to fluorescence-labeled collagen with and without intracellular or extracellular protease inhibitors (or both) before tracing the collagen at both intracellular and extracellular spaces. Collagen were internalized by hMSCs and degraded intracellularly in lysosomes. In the presence of protease inhibitors, both intracellular collagen fibril growth and extracellular deposition of collagen fibrils were enhanced. Moreover, protease inhibitors work synergistically with ascorbic acid, a well-known matrix deposition-enhancing reagent, in further enhancing collagen fibril deposition at the extracellular space.

These findings provide a better understanding of the interactions between hMSCs and collagen biomaterials and suggest a method to manipulate matrix remodeling and deposition of hMSCs, contributing to better scaffolding for tissue engineering and regenerative medicine.

Does magnetic resonance imaging identify cytoarchitectonic subtypes of the normal human cerebral cortex? Magnetic Resonance Imaging (MRI) allows a detailed "in vivo" macroscopic study of the human brain; previously, it has been demonstrated that Fluid Attenuated Inversion Recovery (FLAIR) sequence shows higher signal intensity of cortices belonging to limbic structures. To measure and compare signal intensities (SI) of cytoarchitectonically different cortical regions. In 22 adult subjects, without psychiatric or neurological diseases, FLAIR sequence was performed in coronal slices, perpendicular to the main hippocampal axis. Signal intensity was measured, with a region-of-interest (ROI) function, in 12 different cortical regions. We compared these values and grouped the cortices into five groups: (1) limbic cortices, (2) paralimbic agranular cortices, (3) paralimbic granular cortices, (4) parietal-type neopallium, (5) frontal-type neopallium. A t-test for comparison of paired samples was performed, considering p</=0.05 as statistically significant. We found statistically significant differences amongst the different groups, with the exception of groups 1 and 2, which did not show differences between them. No statistically significant differences were found among cortices belonging to the same group.

Structural characteristics of the cerebral cortex cause changes in its signal intensity. Magnetic resonance imaging (FLAIR sequence) allows discrimination of different cytoarchitectonic areas of the human cerebral cortex.

Do core exercises elevate trunk stability to facilitate skilled motor behavior of the upper extremities? The purpose of this study was to investigate the influence of core exercises on upper extremity function relative to skilled motor behavior and postural sway. We examined the effects of core exercises on the skilled motor behavior and postural sway of 40 healthy students who were assigned randomly to the core exercise group or the control group. Independent variable is extent of exposure to core exercise and dependent variables are skilled motor behavior and postural sway. A Purdue pegboard which measures skilled motor behavior and a stabilometer which measures postural sway were used to evaluate the influence of core exercises. Pre-intervention and post-intervention skilled motor behavior and postural sway were compared between the core exercise group and control group using the Wilcoxon rank sum test; a significance level of α = 0.05 was considered statistically significant. Also, we investigated the application of core exercises in a clinical setting for one patient with cerebral vascular disease. The post intervention skilled motor behavior (p = 0.04) and postural sway, LNG (p = 0.05), LNG/TIME (p = 0.04) and X LNG (p = 0.02) were significantly higher in the core exercise group than control group. In the case report, there were good results; function of the upper extremity improved after doing the exercises. There were positive changes in some daily living activities.

Core exercises are likely to enhance trunk stabilization to improve upper extremity function. It is possible for core exercises to be adapted for patients.

Does weekly doxorubicin increase coronary arteriolar wall and adventitial thickness? Doxorubicin (DOX) is associated with premature cardiovascular events including myocardial infarction. This study was performed to determine if the weekly administration of DOX influenced coronary arteriolar medial and/or adventitial wall thickening. Thirty-two male Sprague-Dawley rats aged 25.1± 2.4 weeks were randomly divided into three groups and received weekly intraperitoneal injections of normal saline (saline, n = 7), or low (1.5 mg/kg to 1.75 mg/kg, n = 14) or high (2.5 mg/kg, n = 11) doses of DOX. The animals were treated for 2-12 weeks, and euthanized at pre-specified intervals (2, 4, 7, or 10+ weeks) to obtain histopathologic assessments of coronary arteriolar lumen diameter, medial wall thickness, adventitial wall thickness, and total wall thickness (medial thickness + adventitial thickness). Lumen diameter was similar across all groups (saline: 315±34 µm, low DOX: 286±24 µm, high DOX: 242±27 µm; p = 0.22). In comparison to animals receiving weekly saline, animals receiving weekly injections of 2.5 mg/kg of DOX experienced an increase in medial (23±2 µm vs. 13±3 µm; p = 0.005), and total wall thickness (51±4 µm vs. 36±5 µm; p = 0.022), respectively. These increases, as well as adventitial thickening became more prominent after normalizing for lumen diameter (p<0.05 to p<0.001) and after adjusting for age, weight, and total cumulative DOX dose (p = 0.02 to p = 0.01). Animals receiving low dose DOX trended toward increases in adventitial and total wall thickness after normalization to lumen diameter and accounting for age, weight, and total cumulative DOX dose (p = 0.06 and 0.09, respectively).

In conclusion, these data demonstrate that weekly treatment of rats with higher doses of DOX increases coronary arteriolar medial, adventitial, and total wall thickness. Future studies are warranted to determine if DOX related coronary arteriolar effects are reversible or preventable, exacerbate the known cardiomyopathic effects of DOX, influence altered resting or stress-induced myocardial perfusion, or contribute to the occurrence of myocardial infarction.

Are growth rates of renal cell carcinoma and oncocytoma under surveillance similar? Through examining our experience with renal mass surveillance, we hoped to determine factors suggestive of renal cell carcinoma. We followed for at least 1 year 41 patients with 47 solid renal masses (mean diameter 2.0 cm, range 0.8-5). Mean surveillance duration was 29 months and was more than 2 years for 23 masses (49%). Overall mean increase in diameter was 0.27 cm/year, but 21 (45%) did not grow, and mean growth rate was 0.5 cm/year in the 26 that did grow. Of the masses, 14 have been treated, 33 continue to be followed, and pathology is known in 16 (34%). Growth was seen in all 6 known oncocytomas (mean 0.52 cm/year), 80% of the 10 biopsy proven renal cell carcinomas grew (mean 0.71 cm/year), but only 12 (39%) of the masses with unknown pathology (0.08 cm/year). There was no factor that distinguished oncocytomas from renal cell carcinomas. In 1 patient, a 3-cm mass that had not changed in size for 6 years doubled in size over 6 months, and metastatic disease developed.

Although growth of most renal masses is slow, some grow quickly, and delayed growth with metastases can occur. No factor distinguished renal cell carcinomas from oncocytomas. Surveillance for renal masses remains an option but must be rigorous and continuous, and is not without risk of progression.

Are primary care patients with depression less accepting of treatment than those seen by mental health specialists? This study examined whether depressed patients treated exclusively in primary care report less need for care and less acceptability of treatment options than those depressed patients treated in the specialty mental health setting after up to 6 months of treatment. Cross-sectional study. Forty-five community primary care practices. A total of 881 persons with major depression who had received mental health services in the previous 6 months and who enrolled in 3 of the 4 Quality Improvement for Depression Collaboration Studies. Patients were categorized into 1 of 2 groups: 1) having received mental health services exclusively from a primary care provider (45%), or 2) having received any services from a mental health specialist (55%) in the previous 6 months. Compared with patients who received care from mental health specialists, patients who received mental health services exclusively from primary care providers had 2.7-fold the odds (95% confidence interval [CI], 1.6 to 4.4) of reporting that no treatment was definitely acceptable and had 2.4-fold the odds (95% CI, 1.5 to 3.9) of reporting that evidence-based treatment options (antidepressant medication) were definitely not acceptable. These results were adjusted for demographic, social/behavioral, depression severity, and economic factors using multiple logistic regression analysis.

Patients with depression treated exclusively by primary care providers have attitudes and beliefs more averse to care than those seen by mental health specialists. These differences in attitudes and beliefs may contribute to lower quality depression care observed in comparisons of primary care and specialty mental health providers.

Is serum ferritin an independent predictor of histologic severity and advanced fibrosis in patients with nonalcoholic fatty liver disease? Serum ferritin (SF) levels are commonly elevated in patients with nonalcoholic fatty liver disease (NAFLD) because of systemic inflammation, increased iron stores, or both. The aim of this study was to examine the relationship between elevated SF and NAFLD severity. Demographic, clinical, histologic, laboratory, and anthropometric data were analyzed in 628 adult patients with NAFLD (age, ≥ 18 years) with biopsy-proven NAFLD and an SF measurement within 6 months of their liver biopsy. A threshold SF >1.5 × upper limit of normal (ULN) (i.e., >300 ng/mL in women and >450 ng/mL in men) was significantly associated with male sex, elevated serum alanine aminotransferase, aspartate aminotransferase, iron, transferrin-iron saturation, iron stain grade, and decreased platelets (P < 0.01). Histologic features of NAFLD were more severe among patients with SF >1.5 × ULN, including steatosis, fibrosis, hepatocellular ballooning, and diagnosis of NASH (P < 0.026). On multiple regression analysis, SF >1.5 × ULN was independently associated with advanced hepatic fibrosis (odds ratio [OR], 1.66; 95% confidence interval [CI], 1.05-2.62; P = 0.028) and increased NAFLD Activity Score (NAS) (OR, 1.99; 95% CI, 1.06-3.75; P = 0.033).

A SF >1.5 × ULN is associated with hepatic iron deposition, a diagnosis of NASH, and worsened histologic activity and is an independent predictor of advanced hepatic fibrosis among patients with NAFLD. Furthermore, elevated SF is independently associated with higher NAS, even among patients without hepatic iron deposition. We conclude that SF is useful to identify NAFLD patients at risk for NASH and advanced fibrosis.

Does leptin promote epithelial-mesenchymal transition of breast cancer via the upregulation of pyruvate kinase M2? Accumulating researches have shown that epithelial-mesenchymal transition (EMT) contributes to tumor metastasis. Leptin, a key adipokine secreted from adipocytes, shapes the tumor microenvironment, potentiates the migration of breast cancer cells and angiogenesis, and is also involved in EMT. However, the potential mechanism remains unknown. This study aims to explore the effect of leptin on EMT in breast cancer cells and the underlying mechanism. With the assessment of EMT-associated marker expression in MCF-7, SK-BR-3, and MDA-MB-468 cells, the effect of leptin on breast cancer cells was analyzed. Besides, an array of pathway inhibitors as well as RNA interference targeting pyruvate kinase M2 (PKM2) were used to clarify the underlying mechanism of leptin-mediated EMT in vitro and in vivo. The results demonstrated that leptin promoted breast cancer cells EMT, visibly activated the PI3K/AKT signaling pathway, and upregulated PKM2 expression. An antibody against the leptin receptor (anti-ObR) and the PI3K/AKT signaling pathway inhibitor LY294002 significantly abolished leptin-induced PKM2 expression and EMT-associated marker expression. SiRNA targeting PKM2 partially abolished leptin-induced migration, invasion, and EMT-associated marker expression. In vivo xenograft experiments indicated that RNA interference against PKM2 suppressed breast cancer growth and metastasis.

Our data suggest that leptin promotes EMT in breast cancer cells via the upregulation of PKM2 expression as well as activation of PI3K/AKT signaling pathway, and PKM2 might be one of the key points and potential targets for breast cancer therapy.

Does magnesium lithospermate B reduce myocardial ischemia/reperfusion injury in rats via regulating the inflammation response? Magnesium lithospermate B (MLB), an active polyphenol acid of Danshen [Radix Salviae miltiorrhizae (Labiatae)], showed renoprotective, neuroprotective and myocardial salvage effects. Previous studies demonstrated that MLB could effectively suppress the production of cytokines and their associated signaling pathways in activated human T cells. The purpose of this study was to examine the beneficial effects of MLB on myocardial ischemia/reperfusion (MI/R) injury and to explore its potential mechanisms related to anti-inflammation. Sprague-Dawley rats were grouped as sham group, model group and MLB-treated (15, 30 and 60 mg/kg) groups. Animals were subjected to MI/R injury by the occlusion of left anterior descending artery for 30 min followed by reperfusion for 3 h. At the end of reperfusion, blood samples were collected to determine the serum levels of cardiac troponin (cTnI), creatine kinase-MB (CK-MB), tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β) and interleukin 6 (IL-6). Hearts were harvested to assess infarct size, histopathological changes and the activity of myeloperoxidase (MPO). The expression of phosphor-IkB-α and phosphor-nuclear factor kappa B (NF-κB) were assayed by western blot. MLB administration significantly (p < 0.05) reduced: (1) ST-segment elevation (0.23 mv), (2) the infarct size (22.5%), (3) histological scores of myocardial injury (1.67 score), (4) myocardial injury marker enzymes: cTnI (5.64 ng/ml) and CK-MB (49.57 ng/ml) levels, (5) pro-inflammatory cytokines: TNF-α (97.36 pg/ml), IL-1β (93.35 pg/ml) and IL-6 (96.84 pg/ml) levels, (6) MPO activity (1.82 U/mg), (7) phosphor-NF-κB (0.87) and phosphor-IkB-α (0.96) expression.

Our study provided evidence that MLB ameliorated the inflammatory process associated with MI/R injury via NF-κB inactivation.

Does poly-ADP-ribose-polymerase inhibition ameliorate hind limb ischemia reperfusion injury in a murine model of type 2 diabetes? Diabetes is known to increase poly-ADP-ribose-polymerase (PARP) activity and posttranslational poly-ADP-ribosylation of several regulatory proteins involved in inflammation and energy metabolism. These experiments test the hypothesis that PARP inhibition will modulate hind limb ischemia reperfusion (IR) in a mouse model of type-II diabetes and ameliorate the ribosylation and the activity/transnuclear localization of the key glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). db/db mice underwent 1.5 hours of hind limb ischemia followed by 1, 7, or 24 hours of reperfusion. The treatment group received the PARP inhibitor PJ34 (PJ34) over a 24-hour period; the untreated group received Lactated Ringer (LR) at the same time points. IR muscles were analyzed for indices of PARP activity, fiber injury, metabolic activity, inflammation, GAPDH activity/intracellular localization, and poly-ADP-ribosylation of GAPDH. PARP activity was significantly lower in the PJ34-treated groups than in the Lactated Ringer group at 7 and 24 hours of reperfusion. There was significantly less muscle fiber injury in the PJ34-treated group than in the Lactated Ringer-treated mice at 24 hours of reperfusion. PJ34 lowered levels of select proinflammatory molecules at 7 hours and 24 hours of IR. There were significant increases in metabolic activity only at 24 hours of IR in the PJ34 group, which temporally correlated with increase in GAPDH activity, decreased GAPDH poly-ADP-ribosylation, and nuclear translocation of GAPDH.

PJ34 reduced PARP activity, GAPDH ribosylation, and GAPDH translocation; ameliorated muscle fiber injury; and increased metabolic activity after hind limb IR injury in a murine model of type-II diabetes. PARP inhibition might be a therapeutic strategy after IR in diabetic humans.

Is extended embryo culture associated with increased adverse obstetric or perinatal outcome? We sought to compare obstetric and perinatal outcomes of singletons born after extended embryo culture and a single blastocyst stage embryo vs a single cleavage stage embryo transfer. This was a retrospective cohort study of 1543 fresh single embryo transfers using nondonor oocytes in women ≤40 years old from December 2008 through December 2012 at the reproductive unit of McGill University Health Center. The main outcome measures were perinatal outcomes including birthweight, low birthweight, small for gestational age, preterm delivery, preeclampsia, placental abruption, and neonatal complications. Covariates were maternal age, body mass index, smoking, cause of infertility, parity, and sex of the baby. Transfers of 693 fresh single cleavage embryos and 850 fresh single blastocysts resulting in 564 pregnancies and 381 singleton deliveries were analyzed. Blastocyst transfer resulted in a higher clinical pregnancy rate (50.1% and 19.9%) and live birth rate (33.5% and 13.8%) compared to cleavage embryo transfer, respectively (P < .001). Multivariate analyses for pregnancy revealed no increased risk of maternal or neonatal complications in pregnancies resulting from extended embryo culture.

Live births resulting from extended embryo culture and a single blastocyst transfer are not associated with increased adverse obstetric and perinatal outcome compared to live births from a single cleavage embryo transfer in women ≤40 years old.

Is initial metastatic kinetics the best prognostic indicator in stage IV metastatic melanoma? In metastatic melanoma (MM) there is an agreement that a fast or slow progression should influence the choice between drugs with immediate impact (BRAF-inh) or delayed (ipilimumab) activity. MM kinetics thus appears crucial for medical decision, although only estimated through surrogate markers (tumour load or lactate dehydrogenase (LDH)). Our objective was to show that 1-MM kinetics can be measured and 2- is a real prognostic factor. Among all stage IV MM, we retrospectively select those with long follow-up who had two comparable total body computed tomography (CT) scans within the first 3 months, and did not receive meantime any treatment with a likely impact on MM kinetics. Kinetics index (KI) was calculated from changes in total metastatic volume (ΔTMV/ΔT). In 126 patients, KI of progression ranges from 0 to 24,839 mm3/day. Overall survival (OS) was significantly much lower in the higher terciles of KI than in the lower ones (median OS of 459, 388 and 183 days, for KI of 0-99, 100-999 and > or =1000 mm3/day, respectively). In the multivariate analysis, KI was more predictive of OS than LDH or tumour load.

Delaying major treatments in stage IV MM for a few weeks permits a measure of KI, which is the best prognostic indicator in MM. The huge range of KI probably reflects major differences in aggressiveness that any therapeutic decision should take into account. KI could be used to assess prospectively how much the efficacy of each new MM drugs is influenced by MM initial kinetics.

Is positron emission tomography scanning superior to whole body multidetector helical computed tomography in the preoperative staging of colorectal cancer? The role of positron emission tomography with the glucose analogue [18F] fluoro-2-deoxy-D-glucose (FDG-PET) in the initial staging of disease in patients with primary colorectal cancer (CRC) has not been adequately assessed. To evaluate the additional value of FDG-PET as a staging modality, complementary to routine multidetector row computed tomography (MDCT) in patients with CRC. Forty four patients with CRC underwent preoperative MDCT and FDG-PET. The accuracy of intraoperative macroscopic staging was also investigated compared with histopathological diagnosis. All FDG-PET images were evaluated with respect to detectability of the primary tumour, lymph node involvement, and distant metastases. Both MDCT and FDG-PET diagnoses and treatment plan were compared with surgical and histopathological results. Thirty seven patients underwent surgery. Tumour detection rate was 95% (42/44) for MDCT, 100% (44/44) for FDG-PET, and 100% (37/37) for intraoperative macroscopic diagnosis. Pathological diagnosis of T factor was T1 in five, T2 in four, T3 in 24, and T4 in four cases. Concordance rate with pathological findings of T factor was 57% (21/37) for MDCT and 62% (23/37) for macroscopic diagnosis. Lymph node involvement was pathologically positive in 19 cases. Regarding N factor, overall accuracy was 62% (23/37) for MDCT, 59% (22/37) for FDG-PET, and 70% (26/37) for macroscopic diagnosis. For all 44 patients, FDG-PET findings resulted in treatment changes in only one (2%) patient.

FDG-PET is not superior to routine MDCT in the initial staging of primary CRC.

Does host response to translocated microbial products predict outcomes of patients with HBV or HCV infection? Chronic infection with hepatitis B or C virus (HBV or HCV) is a leading cause of cirrhosis by unknown mechanisms of pathogenesis. Translocation of gut microbial products into the systemic circulation might increase because of increased intestinal permeability, bacterial overgrowth, or impaired clearance of microbial products by Kupffer cells. We investigated whether the extent and progression of liver disease in patients with chronic HBV or HCV infection are associated with microbial translocation and subsequent activation of monocytes. In a retrospective study, we analyzed data from 16 patients with minimal fibrosis, 68 with cirrhosis, and 67 uninfected volunteers. We analyzed plasma levels of soluble CD14 (sCD14), intestinal fatty acid binding protein, and interleukin-6 by enzyme-linked immunosorbent assay, and lipopolysaccharide (LPS) by the limulus amebocyte lysate assay, at presentation and after antiviral treatment. Compared with uninfected individuals, HCV- and HBV-infected individuals had higher plasma levels of LPS, intestinal fatty acid binding protein (indicating enterocyte death), sCD14 (produced upon LPS activation of monocytes), and interleukin-6. Portal hypertension, indicated by low platelet counts, was associated with enterocyte death (P=.045 at presentation, P<.0001 after therapy). Levels of sCD14 correlated with markers of hepatic inflammation (P=.02 for aspartate aminotransferase, P=.002 for ferritin) and fibrosis (P<.0001 for γ-glutamyl transpeptidase, P=.01 for alkaline phosphatase, P<.0001 for α-fetoprotein). Compared to subjects with minimal fibrosis, subjects with severe fibrosis at presentation had higher plasma levels of sCD14 (P=.01) and more hepatic CD14+ cells (P=.0002); each increased risk for disease progression (P=.0009 and P=.005, respectively).

LPS-induced local and systemic inflammation is associated with cirrhosis and predicts progression to end-stage liver disease in patients with HBV or HCV infection.

Does eye-tracking reveal a slowdown of social context processing during intention attribution in patients with schizophrenia? Schizophrenia is associated with poor theory of mind (ToM), particularly in the attribution of intentions to others. It is also associated with abnormal gaze behaviours and contextual processing. This study investigated to what extent impaired ToM in patients with schizophrenia is related to abnormal processing of social context. We evaluated ToM using a nonverbal intention attribution task based on comic strips depicting social/nonsocial and contextual/noncontextual events while eye movements were recorded. Eye-tracking was used to assess processing time dedicated to visual cues contained in regions of interest identified in a pilot study. We measured cognitive contextual control on a separate task. We tested 29 patients with schizophrenia and 29 controls. Compared with controls, patients were slower in intention attribution but not in physical reasoning. They looked longer than controls at contextual cues displayed in the first 2 context pictures of the comic strips, and this difference was greater for intention attribution than for physical reasoning. We found no group difference in time spent looking at noncontextual cues. Patients' impairment in contextual control did not explain their increased reaction time and gaze duration on contextual cues during intention attribution.

Difficulty may not have been equivalent between intention attribution and physical reasoning conditions.

Is the correlation between muscle and nerve fiber conduction velocities in thenar muscle lost in case of carpal tunnel syndrome? This study investigated the relationship between muscle fiber conduction velocity (MFCV) and motor nerve conduction velocity (MNCV) in thenar muscle of 20 normal subjects and of 20 patients suffering from a moderate carpal tunnel syndrome. Our goal was to confirm the positive correlation between MFCV and MNCV and to assess the influence of carpal tunnel syndrome on this relationship. MFCV was calculated in voluntarily contracted thenar muscle using a multi-channel surface recording and a spike-triggered averaging technique. MFCV values ranged between 2.6 and 7.2 m/s (mean+/-SEM: 4.5+/-0.3) in normal subjects and between 3.5 and 6.9 m/s (4.7+/-0.2) in patients. Subjects and patients did not differ regarding MFCV values, but a correlation between MFCV and MNCV was found in normal subjects (P=0.0005) and not in patients (P=0.54).

A correlation between muscle and nerve conduction velocities existed in healthy subjects but was lost in case of moderate carpal tunnel syndrome. MFCV appeared to be insensitive to focal nerve conduction slowing.

Is mouse mammary tumour virus-like virus ( MMTV-LV ) present within the liver in a wide range of hepatic disorders and unrelated to nuclear p53 expression or hepatocarcinogenesis? The human equivalent of mouse mammary tumour virus (MMTV-LV) may have a role in the pathogenesis of primary biliary cirrhosis (PBC) and breast carcinogenesis. We investigated MMTV-LV prevalence in patients with liver disease of diverse aetiology and associations with hepatic expression of hormonal receptors, p53 protein and complicating hepatocarcinogenesis. Nested PCR for MMTV-LV envelope was performed using tissue from 210 patients with liver disease and 20 patients with histologically normal liver. Hepatic expression of estrogen receptor-alpha (ER-alpha), progesterone receptor (PgR) and p53 protein was determined by immunohistochemistry. MMTV-LV expression was also determined in hepatocellular carcinoma (HCC) tissue from 21 patients, in 11 of whom paired non-tumourous liver tissue was available for comparison. MMTV-LV envelope sequences were present in 25% (53/210) of liver disease biopsies but not in histologically normal liver (0/20) (P=0.02). There was no association between MMTV-LV presence and hepatic expression of ER-alpha or p53 protein. No liver sections were PgR positive. MMTV-LV prevalence was not significantly different in HCC tissue, paired non-tumourous chronic liver disease tissue from the same patients and liver disease tissue from patients without complicating HCC.

Hepatic expression of MMTV-LV is evident in a wide range of non-cirrhotic and cirrhotic liver diseases, irrespective of ER-alpha, PgR or p53 status, and unrelated to complicating HCC.

Is high blood eosinophil count associated with more frequent asthma attacks in asthma patients? The clinical importance of eosinophils in asthma has been shown by the observation of frequent exacerbation in patients with high sputum eosinophil counts and a corresponding decrease in exacerbations when anti-inflammatory therapy was adjusted to maintain low sputum eosinophil percentages. However, less is known of the relation between blood eosinophilia and asthma exacerbation. To examine whether patients with asthma and a higher blood eosinophil count have more asthma attacks than those with a lower count. The authors analyzed data from the National Health and Nutrition Examination Survey, an annual cross-sectional survey of the US general population. Patients with asthma and asthma attacks were identified based on participants' self-report or parental report. A high blood eosinophil count was defined using 200, 300, or 400 cells/μL as cutoffs. The primary analysis used data from 2001 through 2010 after adjusting for demographic variables, obesity, smoking, neutrophil level, and past treatment for wheezing. A secondary analysis used data from 2007 through 2010 and included recent treatment for asthma and fraction of exhaled nitric oxide level as additional adjustment variables. In survey years 2001 through 2010, 3,162 patients with asthma had blood eosinophil data and approximately half (54% of children and 52% of adults) reported an asthma attack in the previous year. In the primary analysis, higher blood eosinophil counts were associated with more asthma attacks in children but not in adults. The secondary analysis suggested an association in both children and adults.

Patients with asthma with higher blood eosinophil counts experienced more asthma attacks than those with lower eosinophil counts.

Does niemann-Pick C1 modulate hepatic triglyceride metabolism and its genetic variation contributes to serum triglyceride levels? To study how Niemann-Pick disease type C1 (NPC1) influences hepatic triacylglycerol (TG) metabolism and to determine whether this is reflected in circulating lipid levels. In Npc1(-/-) mice, the hepatic cholesterol content is increased but the TG content is decreased. We investigated lipid metabolism in Npc1(-/-) mouse hepatocytes and the association of NPC1 single-nucleotide polymorphisms with circulating TGs in humans. TGs were reduced in Npc1(-/-) mouse serum and hepatocytes. In Npc1(-/-) hepatocytes, the incorporation of [3H]oleic acid and [3H]acetate into TG was decreased, but shunting of oleic acid- or acetate-derived [3H]carbons into cholesterol was increased. Inhibition of cholesterol synthesis normalized TG synthesis, content, and secretion in Npc1(-/-) hepatocytes, suggesting increased hepatic cholesterol neogenesis as a cause for the reduced TG content and secretion. We found a significant association between serum TG levels and 5 common NPC1 single-nucleotide polymorphisms in a cohort of 1053 men, with the lowest P=8.7 x 10(-4) for the single-nucleotide polymorphism rs1429934. The association between the rs1429934 A allele and higher TG levels was replicated in 2 additional cohorts, which included 8041 individuals.

This study provides evidence of the following: (1) in mice, loss of NPC1 function reduces hepatocyte TG content and secretion by increasing the metabolic flux of carbons into cholesterol synthesis; and (2) common variation in NPC1 contributes to serum TG levels in humans.

Do the majority of surgical departments adhere to national Danish guidelines for surveillance after colorectal cancer surgery? In 2003 the use of post-operative surveillance (POS) after surgery for colorectal cancer (CRC) in Denmark was studied. Diversity in the choice and frequency of surveillance modalities was found. Subsequently, the Danish Colorectal Cancer Group (DCCG) has published guidelines for POS. In the same period, the number of departments performing CRC surgery has been reduced by 50% nationally. The aim of the present study was to describe the POS after CRC in Denmark following a reduction in the number of departments performing operations for CRC and the DCCG's publication of national recommendations for POS programmes. Questionnaires were sent to all 19 departments that performed operations for CRC. Questions concerned the diagnostic modalities used for detecting recurrences and metachrone cancers. All departments returned their questionnaires. All departments had a formal POS programme. The recommendations given by the DCCG were met by 17 departments (89%) with regard to liver metastases, by 16 departments (84%) with regard to lung metastases and by 16 departments (84%) with regard to metachrone cancers.

As opposed to what was observed in 2003, all departments offered a POS programme after CRC surgery in 2012. Almost all departments met the DCCG recommendations, probably owing to the centralization of CRC surgery and the DCCG's introduction of national guidelines. Hopefully, this will contribute to a better survival for CRC patients in the future, although more research is needed to establish optimal post-operative surveillance.

Does [ Cervical esophagostomy improve the life quality of patients with dysphagia induced by radiotherapy for nasopharyngeal carcinoma ]? To evaluate the effect of cervical esophagostomy for the treatment of patients with dysphagia induced by radiotherapy, in order to improve the therapeutic effects. A retrospective study was performed on 53 nasopharyngeal carcinoma (NPC) patients with dysphagia, who received cervical esophagostomy. The nutritional status of these patients was measured at five given time before and after operation. The occurrence of pneumonia and reflux esophagitis before and after operation was recorded, and the quality of life based on SF-36 quality of life (QOL) scale was studied. After operation, the nutritional status of these patients improved substantially, including the weight, levels of hemoglobin, total protein, albumin and transferring (P<0.05). The pneumonia-infection decreased from 60.38% (32/53) before operation to 15.22% (7/46) after operation (χ(2)=21.04, P<0.01). The incidences of reflux esophagitis decreased from 26.42% (14/53)without operation to 6.52% (3/46) after operation (χ(2)=5.00, P<0.01). Meanwhile, the status of physical health, mental health as well as physical function and social function of these patients were improved significantly at 1 month, 6 months, 1 year and 2 years after operation (P<0.05).

Cervical esophagostomy can improve the life quality of patients with dysphagia induced by radiotherapy for nasopharyngeal carcinoma.

Are paper gestational age wheels generally inaccurate? To compare the estimated date of confinement of paper gestational wheels to the estimated date of confinement of APPs wheels using a standard last menstrual period. Obstetric providers were asked for their gestational wheels. The last menstrual period was set at Jan. 1, 2013, and the estimated date of confinement obtained was compared with the estimated date of confinement of Oct. 8th if the pregnancy completed 280 days. The process was performed on 20 electronic APPs downloadable to cell phones. The process was repeated for both for the leap year of 2012. Thirty-one paper wheels from a variety of sources were collected. Ten wheels (35%) were consistent with the standard pregnancy duration of 280 days. Among the wheels surveyed, the largest discrepancy was 4 days short of 280 days. Two wheels gave an estimated date of confinement that differed from each other by 7 days. Wheels from the same source did not agree with each other. Twenty electronic gestational age calculators were examined. All 20 gave an estimated date of confinement of Oct. 8 consistent with 280 days. None of the paper gestational wheels but all of the APPs corrected for a leap year.

In contrast to APPs gestational age calculators, the estimated date of confinement of the majority of paper wheels deviated from the standard pregnancy duration of 280 days. Precision in gestational age assessment is critical in a variety of clinical settings and heightened by the focus by payers and reporting agencies on elective deliveries before 39 weeks. The use of paper gestational age wheels should be abandoned.

Does three-dimensional spheroid culture promote odonto/osteoblastic differentiation of dental pulp cells? Three-dimensional (3D) spheroid culture is a method for creating 3D aggregations of cells and their extracellular matrix without a scaffold mimicking the actual tissues. The aim of this study was to evaluate the effects of 3D spheroid culture on the phenotype of immortalized mouse dental papilla cells (MDPs) that have the ability to differentiate into odontoblasts. We cultured MDPs for 1, 3, 7, and 14 days in 96-well low-attachment culture plates for 3D spheroid culture or flat-bottomed plates for two-dimensional (2D) monolayer culture. Cell proliferation and apoptosis were detected by immunohistochemical staining of Ki67 and cleaved caspase-3, respectively. Hypoxia was measured by the hypoxia probe LOX-1. Odonto/osteoblastic differentiation marker gene expression was evaluated by quantitative PCR. We also determined mineralized nodule formation, alkaline phosphatase (ALP) activity, and dentine matrix protein-1 (DMP1) expression. Vinculin and integrin signalling-related proteins were detected immunohistochemically. Odonto/osteoblastic marker gene expression and mineralized nodule formation were significantly up-regulated in 3D spheroid-cultured MDPs compared with those in 2D monolayer-cultured MDPs (p<0.05). Histologically, 3D spheroid colonies consisted of two compartments: a cell-dense peripheral zone and cell-sparse core zone. Proliferating cells with high ALP activity and DMP1 expression were found mainly in the peripheral zone that also showed strong expression of vinculin and integrin signalling-related proteins. In contrast, apoptotic and hypoxic cells were detected in the core zone.

3D spheroid culture promotes odonto/osteoblastic differentiation of MDPs, which may be mediated by integrin signalling.

Does ultrasound provide reliable results for the measurement of the patellar tendon cross sectional area? This study examined the reliability of patellar tendon cross sectional area (CSA) measurement using brightness mode ultrasonography. The patellar tendon CSA of fourteen participants was examined on two different days and at three different positions (proximal, median and distal). Five trials per day were conducted in each position, replacing the ultrasound probe on every trial. The images were examined by three different and equally experienced observers. We compared the mean of the five trials in each position to examine the day, observer and position effect. Further, Bland and Altman plots, root mean square (RMS) differences and intraclass correlation coefficients (ICC) were calculated. There was a significant (p < 0.05) day, observer and position effect on the CSA, while the average ICC was 0.592. The Bland and Altman plots showed that differences between conditions or groups, should be in average lower than 37% or higher than 55% of the patellar tendon CSA to be important for clinical or intervention studies.

Our findings show low reliability of the method, which resulted from the low clarity and unclear visibility of tissue boundaries in the ultrasound images. Therefore, the measurement of the CSA of the patellar tendon using ultrasound does not provide accurate and reliable results.

Does epidermal Growth Factor promote Proliferation and Migration of Follicular Outer Root Sheath Cells via Wnt/β-Catenin Signaling? To investigate the effect and molecular mechanism of EGF on the growth and migration of hair follicle outer root sheath (ORS) cells. Intact anagen hair follicles were isolated from mink skin and cultured with EGF in vitro to measure ORS daily growth. Meanwhile, purified primary ORS cells were treated or transfected with EGF, and their proliferation and migration were assessed by MTT assay and transwell assay, respectively. The signaling pathway downstream of EGF was characterized by using the Wnt/β-catenin signaling inhibitor, XAV-939. EGF of 2-20 ng/ml, not higher or lower, promoted the growth of follicular ORS in vitro. EGF treatment or overexpression promoted the proliferation and migration of ORS cells. Moreover, EGF stimulation induced nuclear translocation of β-catenin, and upregulated the expression of Wnt10b, β-catenin, EGF receptor and SOX9. Inhibition of Wnt/β-catenin signaling by XAV-939 significantly reduced the basal and EGF-enhanced proliferation and migration of ORS cells. In addition, a number of follicle-regulatory genes, such as Survivin, Msx2 and SGK3, were upregulated by EGF in the ORS cells, which was also inhibited by XAV-939.

EGF promotes the proliferation and migration of ORS cells and modulates the expression of several follicle-regulatory genes via Wnt/β-catenin signaling.

Does matrix Metalloproteinase 14 Overexpression be Correlated with the Progression and Poor Prognosis of Nasopharyngeal Carcinoma? Matrix metalloproteinase 14 (MMP14) has been identified to play a significant role in several types of cancers, but little is known about the significance of MMP14 in nasopharyngeal carcinoma (NPC) patients. The aim of this study was to explore the association of MMP14 expression with clinicopathologic features and prognosis in NPC. MMP14 mRNA and protein expressions were examined in NPC and nasopharyngeal tissues through real-time PCR and immunohistochemistry. Meanwhile, the relationship of MMP14 expression levels with clinical features and prognosis of NPC patients was analyzed. MMP14 mRNA expression was markedly higher in NPC tissues than in nasopharyngeal epithelium tissues (p = 0.002). Using immunohistochemistry, staining for MMP14 protein was found in the normal nasopharyngeal epithelial cells and malignant epithelial cells, but increased expression of MMP14 was observed in NPC samples compared with normal nasopharyngeal epithelium samples (p = 0.027). In addition, high levels of MMP14 protein were positively correlated with the status of clinical stage (p = 0.009), N classification (p = 0.006), and distant metastasis (p = 0.005) of NPC patients. Patients with higher MMP14 expression had a significantly shorter overall survival time than did patients with low MMP14 expression. Multivariate analysis indicated that the level of MMP14 expression was an independent prognostic indicator (p < 0.001) for the survival of patients with NPC.

MMP14 overexpression is a potentially unfavorable prognostic factor for NPC patients.

Does a mutation in the centriole-associated protein centrin cause genomic instability via increased chromosome loss in Chlamydomonas reinhardtii? The role of centrioles in mitotic spindle function remains unclear. One approach to investigate mitotic centriole function is to ask whether mutation of centriole-associated proteins can cause genomic instability. We addressed the role of the centriole-associated EF-hand protein centrin in genomic stability using a Chlamydomonas reinhardtii centrin mutant that forms acentriolar bipolar spindles and lacks the centrin-based rhizoplast structures that join centrioles to the nucleus. Using a genetic assay for loss of heterozygosity, we found that this centrin mutant showed increased genomic instability compared to wild-type cells, and we determined that the increase in genomic instability was due to a 100-fold increase in chromosome loss rates compared to wild type. Live cell imaging reveals an increased rate in cell death during G1 in haploid cells that is consistent with an elevated rate of chromosome loss, and analysis of cell death versus centriole copy number argues against a role for multipolar spindles in this process.

The increased chromosome loss rates observed in a centrin mutant that forms acentriolar spindles suggests a role for centrin protein, and possibly centrioles, in mitotic fidelity.

Is volumetric bone mineral density of the tibia increased in subjects with radiographic knee osteoarthritis? Radiographic osteoarthritis (ROA) has previously been shown to be associated with an increase in areal bone mineral density (BMD) as assessed by dual energy X-ray absorptiometry (DXA). Here we have assessed volumetric bone density, size and strength by peripheral quantitative computed tomography (pQCT) in a large population-based cohort study in which knee radiographs were available. Two hundred and ninety-five men and 288 women from the MRC Hertfordshire Cohort Study underwent weight bearing extended knee X-rays and bone density measurement of the ipsi-lateral knee using pQCT. Increasing radiographic grade in men but not women was associated with an increase in tibial total area at 38% site and cortical area at 14% site, but not with volumetric BMD. The tibial fracture loads as well as tibial polar strength strain index at 38% site were also increased. There were no significant associations of tibia bone area, BMD or strength with radiographic grade in women.

ROA is not associated with an increase in volumetric BMD as assessed by pQCT. It is, however, associated with a significant increase in bone area and strength, indicating that the association between ROA and areal BMD is mediated through bone size rather than volumetric BMD.

Does valproic acid ( VPA ) inhibit the epithelial-mesenchymal transition in prostate carcinoma via the dual suppression of SMAD4? The epithelial-mesenchymal transition (EMT) plays an important role in cancer metastasis. Previous studies have reported that valproic acid (VPA) suppresses prostate carcinoma (PCa) cell metastasis and down-regulates SMAD4 protein levels, which is the key molecule in TGF-β-induced EMT. However, the correlation between VPA and the EMT in PCa remains uncertain. Markers of the EMT in PCa cells and xenografts were molecularly assessed after VPA treatment. The expression and mono-ubiquitination of SMAD4 were also analyzed. After transfection with plasmids that express SMAD4 or short hairpin RNA for SMAD4 down-regulation, markers of EMT were examined to confirm whether VPA inhibits the EMT of PCa cells through the suppression of SMAD4. VPA induced the increase in E-cadherin (p < 0.05), and the decrease in N-cadherin (p < 0.05) and Vimentin (p < 0.05), in PCa cells and xenografts. SMAD4 mRNA and protein levels were repressed by VPA (p < 0.05), whereas the level of mono-ubiquitinated SMAD4 was increased (p < 0.05). SMAD4 knockdown significantly increased E-cadherin expression in PC3 cells, but SMAD4 over-expression abolished the VPA-mediated EMT-inhibitory effect.

VPA inhibits the EMT in PCa cells via the inhibition of SMAD4 expression and the mono-ubiquitination of SMAD4. VPA could serve as a promising agent in PCa treatment, with new strategies based on its diverse effects on posttranscriptional regulation.

Is initial cognitive decline associated with cortical thinning in early Parkinson disease? Our aim was to assess cortical thickness in a large multicenter cohort of drug-naive patients with early Parkinson disease (PD), with and without mild cognitive impairment (MCI), and explore the cognitive correlates of regional cortical thinning. One hundred twenty-three newly diagnosed patients with PD and 56 healthy controls with 3-tesla structural MRI scans and complete neuropsychological assessment from the Parkinson's Progression Markers Initiative were included. Modified Movement Disorders Society Task Force level II criteria were applied to diagnose MCI in PD. FreeSurfer image processing and analysis software was used to measure cortical thickness across groups and the association with cognitive domains and tests. In patients with MCI, atrophy was found in temporal, parietal, frontal, and occipital areas compared with controls. Specific regional thinning in the right inferior temporal cortex was also found in cognitively normal patients. Memory, executive, and visuospatial performance was associated with temporoparietal and superior frontal thinning, suggesting a relationship between cognitive impairment and both anterior and posterior cortical atrophy in the whole patient sample.

These findings confirm that MCI is associated with widespread cortical atrophy. In addition, they suggest that regional cortical thinning is already present at the time of diagnosis in patients with early, untreated PD who do not meet the criteria for MCI. Together, the results indicate that cortical thinning can serve as a marker for initial cognitive decline in early PD.

Do ovarian Hormones Regulate SP-D Expression in the Mouse Uterus During Estrous Cycle and Early Pregnancy? Differential expression of SP-D in the cycling human and mouse endometrium suggests its regulation by ovarian hormones. SP-D expression in the mouse uterus was analyzed across the estrous cycle and during early pregnancy. Effect of exogenous ovarian hormones on the uterine expression of SP-D was analyzed. SP-D expression varied across the estrous cycle and peaked in the estrous phase. SP-D transcript levels increased by fourfold in the uteri of estrogen-treated mice while co-administration of estrogen and progesterone enhanced SP-D levels by ninefold. However, treatment with progesterone alone significantly downregulated SP-D expression. Diethylstilbestrol enhanced SP-D transcript levels in the uteri of immature mice by 10-fold. During pregnancy, SP-D levels declined rapidly from 0.5 dpc to 6.5 dpc. In silico analysis predicted the presence of two potential ERE and 1 PRE in the mouse SP-D gene promoter region.

Estrogen positively regulates expression of SP-D in the mouse uterus. Progesterone, along with estrogen synergizes SP-D expression, however, when administered alone results in negative regulation.

Do intratracheally administered titanium dioxide or carbon black nanoparticles aggravate elastase-induced pulmonary emphysema in rats? Titanium dioxide (TiO₂) and carbon black (CB) nanoparticles (NPs) have biological effects that could aggravate pulmonary emphysema. The aim of this study was to evaluate whether pulmonary administration of TiO₂ or CB NPs in rats could induce and/or aggravate elastase-induced emphysema, and to investigate the underlying molecular mechanisms. On day 1, Sprague-Dawley rats were intratracheally instilled with 25 U kg⁻¹ pancreatic porcine elastase or saline. On day 7, they received an intratracheal instillation of TiO₂ or CB (at 100 and 500 μg) dispersed in bovine serum albumin or bovine serum albumin alone. Animals were sacrificed at days 8 or 21, and bronchoalveolar lavage (BAL) cellularity, histological analysis of inflammation and emphysema, and lung mRNA expression of heme oxygenase-1 (HO-1), interleukin-1β (IL-1β), macrophage inflammatory protein-2, monocyte chemotactic protein-1, and matrix metalloprotease (MMP)-1, and -12 were measured. In addition, pulmonary MMP-12 expression was also analyzed at the protein level by immunohistochemistry. TiO₂ NPs per se did not modify the parameters investigated, but CB NPs increased perivascular/peribronchial infiltration, and macrophage MMP-12 expression, without inducing emphysema. Elastase administration increased BAL cellularity, histological inflammation, HO-1, IL-1β and macrophage MMP-12 expression and induced emphysema. Exposure to TiO₂ NPs did not modify pulmonary responses to elastase, but exposure to CB NPs aggravated elastase-induced histological inflammation without aggravating emphysema.

TiO₂ and CB NPs did not aggravate elastase-induced emphysema. However, CB NPs induced histological inflammation and MMP-12 mRNA and protein expression in macrophages.

Is urinary albumin excretion rate associated with increased ambulatory blood pressure in normoalbuminuric type 2 diabetic patients? To evaluate the 24-h blood pressure profile in normoalbuminuric type 2 diabetic patients. A cross-sectional study was conducted in 90 type 2 diabetic patients with a urinary albumin excretion rate (UAER) <20 microg/min on two occasions, 6 months apart (immunoturbidimetry). Patients underwent clinical and laboratory evaluations. Ambulatory blood pressure monitoring and echocardiograms were also performed. UAER was found to correlate positively with systolic doctor's office blood pressure measurements (r = 0.243, P = 0.021) and ambulatory blood pressure (24 h: r = 0.280, P = 0.008) and left ventricular posterior wall thickness (r = 0.359, P = 0.010). Patients were divided into four groups according to UAER (<5, > or =5-10, > or =10-15, and > or =15-20 microg/min). Systolic blood pressure parameters for the 1st, 2nd, 3rd, and 4th groups, respectively, were 123.0 +/- 10.6, 132.5 +/- 15.0, 139.0 +/- 23.4, and 130.7 +/- 8.0 mmHg for 24-h blood pressure (ANOVA P = 0.004) and 48.4 +/- 6.0, 54.5 +/- 11.2, 58.8 +/- 15.6, and 57.6 +/- 8.0 mmHg for 24-h pulse pressure (ANOVA P = 0.003). A progressive increase in the prevalence of diabetic retinopathy was observed from the 1st to the 4th UAER group: 27.3, 43.8, 45.5, and 66.7% (P = 0.029 for trend).

In type 2 diabetic patients, UAER in the normoalbuminuric range is positively associated with systolic ambulatory blood pressure indexes, left ventricular posterior wall thickness, and diabetic retinopathy, suggesting that intensive blood pressure treatment may prevent diabetes complications in these patients.

Does night-time sleep disturbance correlate with neuropsychiatric impairment in patients with cirrhosis? Sleep-wake disturbances are common in patients with cirrhosis and are generally attributed to the presence of hepatic encephalopathy. To determine the relationship between sleep and neuropsychiatric disturbances in patients with cirrhosis. The study population comprised 87 patients, classified as neuropsychiatrically unimpaired or as having minimal/overt hepatic encephalopathy. Nineteen healthy volunteers served as controls. Validated questionnaires were used to assess sleep quality [Pittsburgh sleep quality index (PSQI)], day-time sleepiness [Epworth sleepiness scale (ESS)] and diurnal preference. Health-related quality of life (H-RQoL) was assessed using the 36-item short form health profile (SF-36v1) and the chronic liver disease questionnaire. Patients slept significantly less well than the healthy volunteers (PSQI score: 8.4 +/- 4.9 vs. 4.6 +/- 2.5, P<0.01) and had more pronounced day-time sleepiness (abnormal ESS: 21 vs. 0%; chi(2)=3.8, P=0.05). No significant relationships were observed between sleep indices and the presence/degree of hepatic encephalopathy. H-RQoL was significantly impaired in the patients (SF-36v1 physical score: 36 +/- 15 vs. 50 +/- 10, P<0.001; SF-36v1 mental score: 46 +/- 11 vs. 50 +/- 10, P<0.01); night-time sleep disturbance was an independent predictor of poor H-RQoL (P<0.01).

Sleep-wake abnormalities are common in patients with cirrhosis; they significantly affect H-RQoL but are not related to the presence of hepatic encephalopathy.

Does improving mortality following emergent surgery in older patients require focus on complication rescue? To determine whether a hospital's ability to rescue patients from major complications underlies variation in outcomes for elderly patients undergoing emergent surgery. Perioperative mortality rates in elderly patients undergoing emergent general/vascular operations are high and vary widely across Michigan hospitals. We identified 23,224 patients undergoing emergent general/vascular surgical procedures at 41 hospitals within the Michigan Surgical Quality Collaborative between 2006 and 2011. Hospitals were ranked by risk- and reliability-adjusted 30-day mortality rates and grouped into tertiles. We stratified patients by age (<75 and ≥75 years). Risk-adjusted major complication and failure-to-rescue (ie, mortality after major complication) rates were determined for each tertile of hospital mortality. Risk-adjusted mortality rates in elderly patients varied 2-fold across all hospitals. Complication rates correlated poorly with mortality. Failure-to-rescue rates, however, were markedly higher in high-mortality hospitals (29% lowest tertile vs 41% highest tertile; P < 0.01). When compared with younger patients, overall failure-to-rescue rates were almost 2-fold greater in the elderly (36.1% ≥75 vs 18.7% <75; P < 0.01).

A hospital's failure to rescue patients from major complications seems to underlie the variation in mortality rates across Michigan hospitals after emergent surgery. Although higher failure-to-rescue rates in the elderly may signify their diminished physiological reserve for surviving critical illness, the wide variation across hospitals also highlights the importance of systems aimed at the early recognition and effective management of major complications in this vulnerable population.

Do aging and the effects of a half marathon on Achilles tendon force-elongation relationship? We aimed to determine whether there are different changes in Achilles tendon (AT) mechanical properties in middle-aged, compared to younger runners that might indicate that tendon fatigue, induced by long-distance running, is age-dependent. 27 middle-aged (50-67 years) and 22 younger (21-29 years) participants ran a 21 km route at their own pace (mean and SD: old: 3.1 ± 0.3 m s Muscle strength decreased significantly (P < 0.05) in both groups immediately post-run (old: 17 %; young: 11 %) and recovered to baseline within 20-28 h post-run. AT stiffness did not change for the younger adults, whereas the middle-aged adults showed a significant (P < 0.05) decrease in AT stiffness (22 %). However, tendon stiffness recovered to baseline 20-28 h post-run. Middle-aged, compared to young adults, demonstrated significantly (P < 0.05) greater stride frequency and number, but no correlations with tendon fatigue changes were determined (R

The results suggest that the plasticity of the AT in response to short-term mechanical loading may be age dependent and that the AT length-tension properties of middle-aged runners may be more vulnerable to change following running compared to younger athletes. However, the observed AT changes in the middle-aged runners dissipated within 20-28 h post-run, suggesting that a tendon viscoelastic recovery mechanism may occur in vivo.

Does oxygen prescribing practice at Waikato Hospital meet guideline recommendations? It is the recommendation of the British Thoracic Society oxygen guidelines and the Waikato Hospital prescribing policy that all supplemental oxygen should be prescribed. The aim of this audit was to evaluate the current oxygen prescribing practices on different specialty wards in the Waikato Hospital. The secondary aim was to evaluate potential harm from oxygen toxicity of the patients whose oxygen was not prescribed appropriately. One hundred and twenty inpatients receiving oxygen therapy were randomly selected between December 2012 and April 2013. Forty patients were selected from each of the respiratory, surgical and other medical subspecialty wards. Their medication charts, clinical records and laboratory results were reviewed regarding their oxygen prescription, smoking history, diagnoses of chronic respiratory diseases and previous documentation of type 2 respiratory failure. In total, 51.7% of all the patients audited had correct oxygen prescriptions: 77.5% of respiratory, 52.5% of surgical and 25% of other medical specialities. Among the 50 patients whose oxygen was not prescribed, many were classified as having high risk of potential complications of oxygen toxicity: 44% having known chronic respiratory disease, 70% having smoking history and 16% having previous type 2 respiratory failure.

Current oxygen prescription rates and practices in Waikato Hospital are not satisfactory and can in turn put patients at risk of oxygen toxicity. There is a significant discrepancy in prescribing practices between specialities. Better education of oxygen prescription is required to raise the awareness and to improve the prescribing practice across the hospital.

Does transcription factor Nrf2 protect the brain from damage produced by intracerebral hemorrhage? Intracerebral hemorrhage (ICH) remains a major medical problem for which there is no effective treatment. Oxidative and cytotoxic damage plays an important role in ICH pathogenesis and may represent a target for treatment of ICH. Recent studies have suggested that nuclear factor-erythroid 2-related factor 2 (Nrf2), a pleiotropic transcription factor, may play a key role in protecting cells from cytotoxic/oxidative damage. This study evaluated the role of Nrf2 in protecting the brain from ICH-mediated damage. Sprague-Dawley rats and Nrf2-deficient or control mice received intracerebral injection of autologous blood to mimic ICH. Sulforaphane was used to activate Nrf2. Oxidative stress, the presence of myeloperoxidase-positive cells (neutrophils) in ICH-affected brains, and behavioral dysfunction were assessed to determine the extent of ICH-mediated damage. Sulforaphane activated Nrf2 in ICH-affected brain tissue and reduced neutrophil count, oxidative damage, and behavioral deficits caused by ICH. Nrf2-deficient mice demonstrated more severe neurologic deficits after ICH and did not benefit from the protective effect of sulforaphane.

Nrf2 may represent a strategic target for ICH therapies.

Is the protective effect of moderate hypothermia during intestinal ischemia-reperfusion associated with modification of hepatic transcription factor activation? Moderate hypothermia throughout intestinal ischemia-reperfusion (IIR) injury reduces multiple organ dysfunction. Heat shock proteins (HSPs) have been shown to be protective against ischemia-reperfusion injury, and STAT (Signal Transducers and Activators of Transcription) proteins are pivotal determinants of the cellular response to reperfusion injury. The aim of this study is to investigate the mechanism of hypothermic protection during IIR. Adult rats underwent intestinal ischemia-reperfusion (IIR), 60-minute ischemia and 60-minute reperfusion, or sham (120 minutes) at either normothermia or moderate hypothermia. Four groups of animals were studied: (1) normothermic sham (NS), (2) normothermic IIR (NIIR), (3) hypothermic sham (HS), and (4) hypothermic IIR (HIIR). Western blotting measured heat shock protein expression, phosphorylated (p-) and total (T-) hepatic STAT-1 and STAT-3. There were no differences in expression of HSPs 27, 47, 60, i70, c70, or 90 between any of the experimental groups. NIIR caused a significant increase in p-STAT-1 compared with normothermic sham (P <.05) and a highly significant increase in p-STAT-3 (P <.001), both these increases were completely abolished by moderate hypothermia (P <.01 v NIIR.)

The protective effect of moderate hypothermia on liver is not mediated by HSP expression at this time-point. Hypothermia may act by decreasing hepatic STAT activation, supporting the potential therapeutic role of moderate hypothermia. Modulation of STAT activation may also provide novel therapeutic targets.

Does large cohort study found a statistically significant association between excessive crying in early infancy and subsequent ear symptoms? The diagnosis of infantile colic is based on excessive crying. However, several causal factors can account for this disconcerting, nonspecific symptom. The main aim of this study was to investigate a possible association between excessive crying during the first 6 months of life and subsequent ear problems. Data from a cohort study of 26 983 Danish children were used. Mothers participated in four telephone interviews and one questionnaire and provided information on crying in the first 6 months of life and ear symptoms at the ages of 6 months, 18 months and 7 years. There was a statistically significant association between excessive crying in infancy and subsequent ear symptoms. A gradual increase in subsequent ear problems was seen with increasing crying time at all the data collection times.

The results of this study suggest a possible link between excessive crying and ear infections. Whether such a link is causal or due to common underlying factors is still unknown. We recommend thorough ear examinations in children with symptoms compatible with infantile colic.

Does dominant negative c-Src inhibit angiotensin II induced activation of NHE3 in OKP cells? Angiotensin II is a potent stimulator of the proximal tubule apical membrane Na/H antiporter, encoded by NHE3. The nonreceptor tyrosine kinase, c-Src, plays a key role in regulation of NHE3 by acidosis in the proximal tubule, and in signaling effects of angiotensin II in vascular smooth muscle. The present studies examined the role of c-Src in mediating angiotensin II-induced NHE3 activation in cultured OKP cells. c-Src was inhibited with herbimycin A, a tyrosine kinase inhibitor, and expression of a dominant negative c-Src, c-SrcK295M. Herbimycin A blocked angiotensin II induced increases in Na/H antiporter activity. In two clonal cell lines expressing vector alone, angiotensin II increased Na/H antiporter activity, while in three clones expressing c-SrcK295M, angiotensin II had no effect. Cyclic AMP and protein kinase A have been proposed to be key mediators in regulation of NHE3 by angiotensin II. 10(-4) M 8-bromo cAMP induced a 40 to 50% inhibition of Na/H antiporter activity in cells expressing c-SrcK295M, similar to that seen in wild-type OKP cells. In addition, cells expressing c-SrcK295M responded normally to 10(-7) M dexamethasone with a 50 to 80% increase in Na/H antiporter activity.

These studies demonstrate that c-Src is required for angiotensin II-induced increases in NHE3 activity. Thus, c-Src plays a key role in antiporter activation by acidosis and angiotensin II.

Does use of a pH-stat strategy during retrograde cerebral perfusion improve cerebral perfusion and tissue oxygenation? Although it is well documented that the use of a pH-stat strategy during hypothermic cardiopulmonary bypass improves cerebral blood flow, an alpha-stat strategy has been almost exclusively used during retrograde cerebral perfusion. We investigated the effects of pH-stat and alpha-stat management on brain tissue blood flow and oxygenation during retrograde cerebral perfusion in a porcine model to determine if the use of a pH-stat strategy during retrograde cerebral perfusion improves brain tissue perfusion. Fourteen pigs were managed by an alpha-stat strategy (alpha-stat group, n = 7) or by a pH-stat strategy (pH-stat group, n = 7) during 120 minutes of hypothermic retrograde cerebral perfusion. Retrograde cerebral perfusion was established through the superior vena cava. Brain tissue blood flow and oxygenation were measured continuously with a laser flowmeter and near infrared spectroscopy, respectively. Brain tissue water content was determined at the end of the experiments. During cooling, brain tissue blood flow was significantly higher with use of the pH-stat strategy than with the alpha-stat strategy (86% +/- 10% versus 40% +/- 3% of baseline). During retrograde cerebral perfusion, brain tissue blood flow was also significantly higher (about three times higher) in the pH-stat group than in the alpha-stat group (15% +/- 4% versus 5% +/- 1% of baseline at 60 minutes of retrograde cerebral perfusion). Tissue oxygen saturation appeared to be higher during retrograde cerebral perfusion in the pH-stat group than in the alpha-stat group. Brain tissue blood flow during rewarming remained significantly higher with the use of pH-stat than with the use of alpha-stat. Brain tissue water contents were similar in both groups.

In our pig model, the use of a pH-stat strategy during retrograde cerebral perfusion significantly improves brain tissue perfusion. Therefore, to improve retrograde cerebral blood flow during retrograde cerebral perfusion, it may be preferable to use a pH-stat strategy, rather than an alpha-stat strategy.

Does expression signature of microRNA-181-a reveal its crucial role in the pathogenesis of paediatric systemic lupus erythematosus? Systemic lupus erythematosus (SLE) is an autoimmune disease manifested by self-reactive antibodies due to failure of selection in both B and T lymphocytes leading to immune intolerance accompanied by increased rate of apoptosis and deficiency in the clearance of the apoptotic cells. Micro RNAs regulate posttranscriptional gene expression and have been recently identified to regulate cellular differentiation, establishing immunological tolerance and are involved in the pathogenesis of several diseases. miR-181-a, expressed in haematopoietic cell lineage, has shown to be an important modulator of B and T cell differentiation, maturation and function. This study aims to identify the expression signature of miR-181-a in the peripheral blood of paediatric SLE and its regulatory effect on the consequent expression of its target gene PCAF. Twenty SLE paediatrics patients, 9 healthy controls and 4 FMF patients were enrolled in this study. The relative expression of miR-181-a, miR-223, PCAF and Hdm2 were performed using quantitative real time PCR. For the first time we show that miR-181-a was significantly downregulated in SLE pediatrics as compared to healthy controls. Furthermore, miR-181-a showed significant difference in its expression among groups with different SLEDAI scores. This special signature of miR-181-a expression is unique to SLE as compared to FMF samples which showed a parallel expression to healthy controls. PCAF was upregulated in SLE patients compared to healthy controls, which has an impact on the ubiquitination of Hdm2 and hence releases p53 leading to the induction of apoptosis.

miR-181-a plays an important role in SLE pathogenesis.

Does ergonomic testing of two different types of handle via virtual reality simulation? Ergonomics in laparoscopic surgery is an unsolved problem. Deficiencies of the instrument handles are well-known and described in several reports and studies. Today, virtual training modules for laparoscopic surgery are available. The aim of this study was to evaluate the ability of a virtual reality (VR) simulator to determine the ergonomic properties of two different laparoscopic instrument handles. Two different types of handles, a ring and an axial handle from Richard Wolf, were used to perform the short clip and cut task of the Xitact 500 LS simulator. The task was repeated every 2 days for a period of 5 weeks. After every trial the volunteers were asked structured questions about their preferences while using the two handles. The axial handle was superior or equal to the ring handle in all criteria. Learning curves over the entire time and day by day were similar. No differences were found for travel distances and error rates, but task times were different for both handles. The subjects preferred the axial handle at the end of the study.

It is possible to determine differences in ergonomics of handle design with a VR trainer. In this study, the Richard Wolf axial handle was superior to the ring handle.

Is lumiracoxib 400 mg once daily comparable to indomethacin 50 mg three times daily for the treatment of acute flares of gout? To demonstrate non-inferiority of lumiracoxib 400 mg once daily (o.d.) compared with indomethacin 50 mg three times daily (t.i.d.) in the treatment of acute gout, and to compare the safety and tolerability of these treatments. In this 1-week, multicentre, randomized, double-blind, double-dummy, active-controlled, parallel-group study, patients with a clinical diagnosis of gout, an acute attack of gout in four or more joints within the 48 h prior to evaluation, and at least moderate pain intensity in the target joint were randomized to treatment with lumiracoxib 400 mg o.d. (n = 118) or indomethacin 50 mg t.i.d. (n = 117). The primary efficacy endpoint was the mean change in pain intensity from baseline over days 2-5, assessed on a 5-point Likert scale, where non-inferiority could be claimed if the lower limit of the confidence interval (CI) was greater than -0.5. The patient's and physician's global assessment of response to treatment, and physician's assessment of tenderness, swelling and erythema of the study joint were also assessed. The estimated difference between treatments for the change from baseline in pain intensity over days 2-5 was -0.004 (95% CI -0.207 to 0.199, P > 0.05), indicating that lumiracoxib 400 mg o.d. had comparable efficacy to indomethacin 50 mg t.i.d. for the primary efficacy variable. There was no significant difference between treatments in any of the secondary efficacy variables. Adverse events were reported by 10.2% of patients treated with lumiracoxib and 22.2% of those receiving indomethacin.

Lumiracoxib is as effective as indomethacin for treatment of acute gout and may have a better safety and tolerability profile.

Does bu Shen Tiao Chong recipe restore diminished ovary reserve through the BDNF pathway? The purpose of this study was to explore the molecular pathway of BSTCR (Bu Shen Tiao Chong recipe) in retrieving diminished ovary reserve (DOR). The DOR model was established through injecting cyclophosphamide and the effect of BSTCR was examined under this background. BSTCR was shown to restore depleted brain-derived neurotrophic factor (BDNF), CDC2, cyclin B, GSH1, and P38 levels as well as impaired oocyte maturation and the higher apoptosis induced in DOR. BSTCR also enhances the response of oocytes to in vitro fertilization, with higher implantation rate, birth rate, and placenta weight.

BSTCR might exert its beneficial role in oocyte maturation and restore DOR through regulating the BDNF pathway. And this pathway itself is probably through the consequence on several serum hormones such as FSH, E2, Inhibin B, etc.

Is dysfunctional elimination syndrome a negative predictor for vesicoureteral reflux? We investigated the likelihood of finding vesicoureteral reflux (VUR) in patients with urinary tract infections (UTIs), accompanied by fever or dysfunctional elimination syndrome (DES). Two hundred consecutive voiding cystourethrograms performed in 1997-2002 for a diagnosis of UTI were reviewed. Fever, DES, and the grade and laterality of VUR were recorded. Patients were stratified into two groups by age to allow for assessment of DES symptoms in the older patient population: <2 years (n=68) and > or =2 years (n=132). Ratios were compared using a two-tailed Fisher's exact test. Of the children> or =2 years old, 64/132 (48%) had VUR. Patients who were non-febrile with DES were less likely than patients who were febrile without DES to have VUR [12/34 (35%) vs 23/34 (68%), P=0.02], whereas the risk of dilating VUR [5/34 (15%) vs 11/34 (32%), P=0.15] and bilateral VUR [4/34 (12%) vs 11/34 (32%), P=0.08] was not statistically different. In febrile patients, the presence of DES was associated with a lower risk of VUR [22/51 (43%) vs 23/34 (68%), P=0.03] and dilating VUR [5/51 (10%) vs 11/34 (32%), P=0.01], but not bilateral VUR [8/51 (16%) vs 11/34 (32%), P=0.11].

Children with non-febrile UTI and DES have a significantly lower risk of having VUR compared to children with febrile UTI and no DES. Among children with a history of UTI, DES is a negative predictor for VUR.

Is a single session of low-intensity exercise sufficient to enhance insulin sensitivity into the next day in obese adults? The purpose of this study was to determine the effect of a relatively modest session of exercise on insulin sensitivity and fatty acid uptake the next day in obese adults. Eleven sedentary obese adults (male/female: 3/8; BMI 37 ± 1 kg/m(2); peak oxygen uptake [VO2peak] 20 ± 1 mL/kg/min) completed three experimental trials. On two of these occasions, subjects exercised to expend 350 kcal in the afternoon. These two exercise trials were identical except for the exercise intensity (50% VO2peak [EX50] and 65% VO2peak [EX65]) and the duration of exercise necessary to expend 350 kcal (EX50 = ≈ 70 min; EX65 = ≈ 55 min). Subjects also completed a control trial (CON), without exercise. The next morning, we measured insulin sensitivity (hyperinsulinemic-euglycemic clamp) and whole-body fatty acid uptake (palmitate rate of disappearance from plasma [Rd]). Exercise increased insulin sensitivity the next day, but whereas the 35% improvement after EX50 compared with CON was statistically significant (P = 0.01), the 20% improvement after EX65 was not (P = 0.17). Despite nearly identical values between CON and EX65 (P = 0.88), systemic fatty acid uptake was lower after EX50 compared with EX65 (P = 0.02), but not quite significant compared with CON (P = 0.07). Importantly, the change in fatty acid uptake after exercise compared with CON was negatively correlated with the change in insulin sensitivity for all trials (r = -0.60, P = 0.003).

A relatively modest single session of exercise in obese adults improved insulin sensitivity the next day, and a reduction in systemic fatty acid uptake in the several hours after exercise may be important for this effect.

Does palmitoleic acid ( n-7 ) increase white adipocytes GLUT4 content and glucose uptake in association with AMPK activation? Palmitoleic acid was previously shown to improve glucose homeostasis by reducing hepatic glucose production and by enhancing insulin-stimulated glucose uptake in skeletal muscle. Herein we tested the hypothesis that palmitoleic acid positively modulates glucose uptake and metabolism in adipocytes. For this, both differentiated 3 T3-L1 cells treated with either palmitoleic acid (16:1n7, 200 μM) or palmitic acid (16:0, 200 μM) for 24 h and primary adipocytes from mice treated with 16:1n7 (300 mg/kg/day) or oleic acid (18:1n9, 300 mg/kg/day) by gavage for 10 days were evaluated for glucose uptake, oxidation, conversion to lactate and incorporation into fatty acids and glycerol components of TAG along with the activity and expression of lipogenic enzymes. Treatment of adipocytes with palmitoleic, but not oleic (in vivo) or palmitic (in vitro) acids, increased basal and insulin-stimulated glucose uptake and GLUT4 mRNA levels and protein content. Along with uptake, palmitoleic acid enhanced glucose oxidation (aerobic glycolysis), conversion to lactate (anaerobic glycolysis) and incorporation into glycerol-TAG, but reduced de novo fatty acid synthesis from glucose and acetate and the activity of lipogenic enzymes glucose 6-phosphate dehydrogenase and ATP-citrate lyase. Importantly, palmitoleic acid induction of adipocyte glucose uptake and metabolism were associated with AMPK activation as evidenced by the increased protein content of phospho(p)Thr172AMPKα, but no changes in pSer473Akt and pThr308Akt. Importantly, such increase in GLUT4 content induced by 16:1n7, was prevented by pharmacological inhibition of AMPK with compound C.

In conclusion, palmitoleic acid increases glucose uptake and the GLUT4 content in association with AMPK activation.

Does high expression of phospho-H2AX predict a poor prognosis in colorectal cancer? Colorectal cancer (CRC) is one of the most common causes of cancer-related deaths worldwide. DNA double-strand breaks (DSBs) are deleterious lesions that can lead to chromosomal anomalies, genomic instability and cancer. The histone H2AX plays an important role in response to DNA damage and phosphorylation of H2AX (p-H2AX) is evidence of DSBs. The aim of this study was to evaluate the clinical significance of p-H2AX expression in CRC. p-H2AX expression in CRC tissues was analyzed by immunohistochemistry and correlated with clinicopathological variables using the chi-square test. The prognostic value of p-H2AX for distant metastasis-free survival (DMFS) and overall survival (OS) was evaluated by Kaplan-Meier estimates and the individual prognostic components were analyzed with Cox regression analysis. A high p-H2AX expression in CRC tissues was associated with tumor stage and perineurial invasion. Furthermore, a high p-H2AX expression was associated with poor DMFS and OS. Cox regression analysis also revealed that p-H2AX was an independent predictor of DMFS and OS.

A high p-H2AX expression in CRC tissues is associated with a more malignant cancer behavior, as well as poor patient survival. p-H2AX may, therefore, be an independent prognostic predictor for CRC, as well as a potential therapeutic target.

Are caregiver characteristics associated with neuropsychiatric symptoms of dementia? To determine whether caregiver characteristics are independently associated with neuropsychiatric symptoms of dementia (NPS) after accounting for patient characteristics. Cross-sectional analysis of data from the Medicare Alzheimer's Disease Demonstration and Evaluation study. Community-dwelling residents in eight U.S. cities. Five thousand seven hundred eighty-eight patients with dementia and their caregivers. Caregivers were asked about the presence of 12 NPS in patients with dementia. Caregiver predictors included age, sex, education, income, marital status, relationship to the patient, whether they lived with patient, number of hours per week spent caregiving, self-reported health, dependency in activities of daily living (ADLs) and instrumental activities of daily living (IADLs), depression, and burden. Multivariate linear regression was used to determine which caregiver characteristics were independently associated with reports of more NPS in patients after controlling for the patient's age, sex, dementia severity, level of ADL dependency, and dementia type. Caregivers were on average 64 years old, 72% female, and 49% were the spouse of the patient (32% wives, 17% husbands). The mean burden score of caregivers was 15 (range 0-32, with higher scores indicating more burden), and 32% had significant depressive symptoms. Patients were on average 79 years old, 60% were female, and most had moderate to severe dementia. The mean number of NPS+/-standard deviation was 4.8+/-2.8. After adjusting for patient characteristics, caregivers who were younger, less educated, more depressed, more burdened, or spent more hours per week giving care reported more NPS in care recipients (all P< or =.005).

Certain caregiver characteristics are associated with NPS, independent of patient characteristics, including dementia severity. Clinicians should consider the dynamics between patients and caregivers when managing NPS. Understanding how different caregiver characteristics influence NPS may help tailor caregiver education and interventions.

Is early anticoagulation associated with reduced mortality for acute pulmonary embolism? Acute pulmonary embolism (PE) may be rapidly fatal if not diagnosed and treated. IV heparin reduces mortality and recurrence of PE, but the relationship between survival and timing of anticoagulation has not been extensively studied. We studied 400 consecutive patients in the ED diagnosed with acute PE by CT scan angiography and treated in the hospital with IV unfractionated heparin from 2002 to 2005. Patients received heparin either in the ED or after admission. Time from ED arrival to therapeutic activated partial thromboplastin time (aPTT) was calculated. Outcomes included in-hospital and 30-day mortality, hospital and ICU lengths of stay, hemorrhagic events on heparin, and recurrent venous thromboembolism within 90 days. In-hospital and 30-day mortality rates were 3.0% and 7.7%, respectively. Patients who received heparin in the ED had lower in-hospital (1.4% vs 6.7%; P = .009) and 30-day (4.4% vs 15.3%; P < .001) mortality rates as compared with patients given heparin after admission. Patients who achieved a therapeutic aPTT within 24 h had lower in-hospital (1.5% vs 5.6%; P = .093) and 30-day (5.6% vs 14.8%; P = .037) mortality rates as compared with patients who achieved a therapeutic aPTT after 24 h. In multiple logistic regression models, receiving heparin in the ED remained predictive of reduced mortality, and ICU admission remained predictive of increased mortality.

We report an association between early anticoagulation and reduced mortality for patients with acute PE. We advocate further study with regard to comorbidities to assess the usefulness of modifications to hospital protocols.

Does effect that an educational program for cystic fibrosis patients and caregivers have on the contamination of home nebulizers? To describe the pathogens found in home nebulizers and in respiratory samples of cystic fibrosis (CF) patients, and to evaluate the effect that a standardized instruction regarding cleaning and disinfection of nebulizers has on the frequency of nebulizer contamination. We included 40 CF patients (22 males), all of whom used the same model of nebulizer. The median patient age was 11.2 ± 3.74 years. We collected samples from the nebulizer mouthpiece and cup, using a sterile swab moistened with sterile saline. Respiratory samples were collected by asking patients to expectorate into a sterile container or with oropharyngeal swabs after cough stimulation. Cultures were performed on selective media, and bacteria were identified by classical biochemical tests. Patients received oral and written instructions regarding the cleaning and disinfection of nebulizers. All determinations were repeated an average of two months later. Contamination of the nebulizer (any part) was detected in 23 cases (57.5%). The nebulizer mouthpiece and cup were found to be contaminated in 16 (40.0%) and 19 (47.5%), respectively. After the standardized instruction had been given, there was a significant decrease in the proportion of contaminated nebulizers (43.5%).

In our sample of CF patients, nebulizer contamination was common, indicating the need for improvement in patient practices regarding the cleaning and disinfection of their nebulizers. A one-time educational intervention could have a significant positive impact.

Does immediate cause of death of demented and non-demented elderly? To investigate the immediate causes of death, in autopsied demented and non-demented elderly. Retrospective clinicopathologic correlations. Acute and intermediate care geriatric hospital. 342 hospitalized demented and non-demented elderly (mean age 84.94 +/- 6.9 years) who underwent consecutive postmortem examinations: 120 demented patients with either vascular dementia (VaD, n = 34), mixed dementia (MD, n = 65) or Alzheimer's disease (AD, n=21) neuropathologically confirmed and 222 nondemented elderly. Primary causes of death were similar in both demented and non-demented patients; the commonest were cardiovascular disease and bronchopneumonia. Cardiac causes of death and especially cardiac failure were more frequent in VaD than in AD or MD (respectively P = 0.027 and 0.005). Dementia was an underlying but never a primary cause of death.

Immediate causes of death are similar in elderly demented and non-demented patients.

Is length of human immunodeficiency virus disease and not immune status a risk factor for development of anal carcinoma? The anal epithelium is subject to dysplastic change in patients with human immunodeficiency virus (HIV). We sought to determine if the duration of HIV disease or the patient's immune status were associated with the development of anal carcinoma. HIV-positive patients diagnosed with anal neoplasms were reviewed. Statistical analysis was performed via an unpaired Student t test and the Fisher exact test. Fourteen patients were identified, 7 with anal intraepithelial neoplasms (group 1) and 7 with anal carcinoma (group 2). Human papillomavirus was detected in 100% of patients in group 1 and in 67% of patients in group 2. There was no significant difference in the level of immunosuppression as assessed by the CD4 counts (266.9 +/- 48.5 vs. 274.7 +/- 92.0 cell/c microl; P = .94) and viral loads (19,243 +/- 18,034 vs. 67,140 +/- 39,570 RNA/mL; P = .29) between groups 1 and 2, respectively. Group 2 had been HIV positive for a significantly longer period of time (12.6 +/- 2.3 y) compared with group 1 (5.9 +/- 2.0 y, P = .05).

The most significant factor for the development of invasive anal carcinoma in patients with HIV is duration of disease. As a result of improved long-term survival secondary to new HIV therapy, anal invasive carcinoma will become an increasing problem.

Does naltrexone affect isoflurane minimum alveolar concentration in cats? To test whether naltrexone, an opioid receptor antagonist, affects the minimum alveolar concentration (MAC) of isoflurane in cats, a species that is relatively resistant to the general anesthetic sparing effects of most opioids. Randomized, crossover, placebo-controlled, blinded experimental design. Six healthy adult cats weighing 4.9 ± 0.7 kg. The cats were studied twice. In the first study, baseline isoflurane MAC was measured in duplicate. The drug (saline control or 0.6 mg kg(-1) naltrexone) was administered IV every 40-60 minutes, and isoflurane MAC was re-measured. In the second study, cats received the second drug treatment using identical methods 2 weeks later. Isoflurane MAC was 2.03 ± 0.12% and was unchanged from baseline following saline or naltrexone administration.

Minimum alveolar concentration was unaffected by naltrexone. Because MAC in cats is unaffected by at least some mu-opioid agonists and antagonists, spinal neurons that are directly modulated by mu-opioid receptors in this species cannot be the neuroanatomic sites responsible for immobility from inhaled anesthetics.

Are mature dendritic cells superior to immature dendritic cells in expanding antigen-specific naive and memory CD8+ T cells? For successful dendritic cell (DC)-based immunotherapy, it is critical to identify the most potent stage of human DCs, including immature DCs (imDCs) and mature DCs (mDCs). imDCs were obtained by culturing monocytes in the presence of GM-CSF and IL-4 for 5- 7 days and imDCs were further cultured for 24-48 h in the presence of TNFalpha, IL-6, IL-1beta and PGE2 to obtain mDCs. Melan-A- and EBV (BRF1) peptides were used and the frequency of antigen-specific CD8+ T cells was assessed using appropriate tetramers. mDCs were potent antigen-presenting cells for the induction and proliferation of antigen-specific naive and memory CD8+ T cells and may overcome regulatory functions that suppress antigen-specific CD8+ T cells.

Our findings that mDCs can efficiently expand antigen-specific naive and memory CD8 + T cells have important implications in the development of vaccination strategies and support the use of antigen-loaded mature DCs in human clinical trials

Does neoadjuvant therapy with interleukin-12-loaded polylactic acid microspheres reduce local recurrence and distant metastases? We previously demonstrated that the intratumoral injection of biodegradable polylactic acid microspheres that were loaded with interleukin (IL)-12 can induce a systemic antitumor immunity. We sought to investigate the clinical potential as neoadjuvant therapy. Mice were inoculated with 5 x 10(7) Line-1 cells subcutaneously. Six days later, a single intratumoral injection of IL-12- or BSA-loaded microspheres were given; 14 days later, autopsy was performed to document metastases. Mice were inoculated with 5 x 10(7) Line-1 cells and 10 days later either treated with IL-12- or BSA-loaded microspheres or resected. Treated tumors were resected 6 days after treatment. Mice were observed 45 days for local recurrence before autopsy. Intratumoral injection of IL-12 microspheres resulted in significant suppression of tumor growth compared with controls (599 +/- 255 mm(3) vs 1591 +/- 372 mm(3); P =.001) and pulmonary metastases (0.4 vs 3.8 nodules per mouse; P =.003). Given before the operation, IL-12-loaded microspheres both decreased the local recurrence rate (100% to 40%) and pulmonary metastases (5.2 vs 0.6 nodules per mouse; P =.06). Earlier resection did not improve local recurrence or distant metastases.

Intratumoral injection of IL-12-loaded polylactic acid microspheres promotes the development of systemic antitumor immunity that can eradicate micrometastases. As a neoadjuvant therapy, this can result in decreased local and distant recurrence.

Does intronic CArG box regulate cysteine-rich protein 2 expression in the adult but not in developing vasculature? An absence of cysteine-rich protein 2 (CRP2) enhances vascular smooth muscle cell (VSMC) migration and increases neointima formation after arterial injury; therefore, CRP2 plays an important role in the response to vascular injury. The goal of the present study was to elucidate the molecular mechanisms that preserve CRP2 expression in the adult vasculature and thus might serve to inhibit the response to injury. We generated a series of transgenic mice harboring potential Csrp2 regulatory regions with a lacZ reporter. We determined that the 12-kb first intron was necessary for transgene activity in adult but not in developing vasculature. Within the intron we identified a 6.3-kb region that contains 2 CArG boxes. Serum response factor preferentially bound to CArG2 box in gel mobility shift and chromatin immunoprecipitation assays; additionally, serum response factor coactivator myocardin factors activated CRP2 expression via the CArG2 box. Mutational analysis revealed that CArG2 box was important in directing lacZ expression in VSMC of adult vessels.

Although CRP2 expression during development is independent of CArG box regulatory sites, CRP2 expression in adult VSMC requires CArG2 element within the first intron. Our results suggest that distinct mechanisms regulate CRP2 expression in VSMC that are controlled by separate embryonic and adult regulatory modules.

Is adipocyte size associated with NAFLD independent of obesity , fat distribution , and PNPLA3 genotype? Adipocyte hypertrophy has been suggested to be causally linked with inflammation and systemic insulin resistance. The aim of the study was to determine whether increased adipocyte size is associated with increased liver fat content due to nonalcoholic fatty liver disease (NAFLD) in humans independent of obesity, fat distribution and genetic variation in the patatin-like phospholipase domain-containing 3 gene (PNPLA3; adiponutrin) at rs738409. One hundred nineteen non-diabetic subjects in a cross-sectional study with a median age of 39 years, mean ± SD BMI of 30.0 ± 5.7 kg m(-2) were studied. Abdominal subcutaneous (SC) adipocyte size, liver fat [proton magnetic resonance spectroscopy ((1) H-MRS)], intra-abdominal (IA), and abdominal SC adipose tissue volumes [magnetic resonance imaging (MRI)] and the PNPLA3 genotype at rs738409 were determined. Univariate and multiple linear regression analysis were used to identify independent predictors of liver fat content. In multiple linear regression analysis, age, gender, BMI, the IA/SC ratio, and PNPLA3 genotype explained 42% of variation in liver fat content. Addition of adipocyte size (P < 0.0001) to the model increased the percent of explanation to 53%. Thus, 21% of known variation in liver fat could be explained by adipocyte size alone.

Increased adipocyte size highly significantly contributes to liver fat accumulation independent of other causes.

Does tSG101 Silencing suppress Hepatocellular Carcinoma Cell Growth by Inducing Cell Cycle Arrest and Autophagic Cell Death? The tumor susceptibility gene 101 (TSG101) was originally identified as a tumor-suppressor gene that mediates many molecular and biological processes, such as ubiquitination, endosomal trafficking, cell survival, and virus budding, but its role in hepatocellular carcinoma (HCC) is currently unknown. We assessed the expression of TSG101 in HCC and paracancerous tissues using qPCR. Then, we used the TSG101-specific siRNA mix to disrupt the expression of TSG101 to investigate the subsequent effect on human hepatoma-7 (Huh7) cells. Western blot was used to detect the protein expression of TSG101 and other molecules. Cell growth assay was performed using CCK8. Transwell assay was used to investigate the migration and invasion ability of Huh7 cells after transfection with of TSG101 siRNA. Flow cytometry was used to estimate the effect of TSG101 knockdown on cell cycle and apoptosis. Confocal laser scanning microscopy was used to observe the actin filaments change and the formation of autophagy. TSG101 was over-expressed in HCC tissues. TSG101 silence was able to suppress Huh7 cell proliferation, migration, and invasion. Furthermore, silencing of TSG101 could induce cell cycle arrest at G1 phase and inhibit the expression of cyclin A and cyclin D, while up-regulating the expression of CDK2. The mechanism might be induction of autophagic cell death and inactivation of Akt and ERK1/2.

TSG101 plays an important role in the development of HCC and may be a target for molecular therapy.

Does [ Inhibition of micro RNA-9 expression promote UV-induced ROS damage in nasopharyngeal carcinoma cells ]? To investigate the effects of down-regulated miR-9 expression on ultraviolet rays (UV)-induced reactive oxygen species (ROS) damage in nasopharyngeal carcinoma (NPC) cells. The NPC cells were transfected with inhibitors of miR-9 by lipofectamine to decrease the expression of miR-9, and the cells transfected with inhibitor control as the control. ROS levels following UV exposure were examined with DCF-DA method and the concentration of glutathione was analyzed via the benzoic acid method; DNA damage and apoptosis also were evaluated. There was significant difference in ROS levels between miR-9 expression-inhibited cells and control cells (26 895 ± 218 vs 15 765 ± 927, t = 39.754, P < 0.001), and also there were significant differences in DNA damage rates (28.0% ± 10.0% vs 23.6% ± 9.2%) and in apoptosis rates (8.0% ± 0.9% vs 4.5% ± 0.8%) following UV exposure between two groups of cells. The miR-9 expression-inhibited cells showed lower level (1.87 ± 0.15) µmol/L of glutathione compared with the control cells (9.85 ± 0.15) µmol/L (t = -48.832, P < 0.001).

Inhibition of miR-9 expression promoted UV-induced ROS damage in nasopharyngeal carcinoma cells.

Does differentiation capacity of endothelial progenitor cells correlate with endothelial function in healthy young men? Endothelial progenitor cells (EPCs) have been assumed to maintain vascular endothelial integrity, so the present study investigated whether the functional capacity of EPCs correlates with endothelial function in healthy young subjects, as has been confirmed in aged subjects with atherosclerotic disease. EPCs in 41 healthy, young male nonsmokers (age 33.1 +/-3.9 years, mean +/- SD) were characterized. The correlation between flow-mediated vasodilation (FMD) and the number of EPCs or the plasma concentrations of growth factors, such as vascular endothelial growth factor, did not reach statistical significance. However, FMD was significantly correlated with the EPC differentiation index, defined as the ratio of the number of EPCs to the total number of adherent cells (r=0.391, P=0.011) and the abundance of endothelial nitric oxide synthase mRNA (r=0.340, P=0.030).

In healthy young men, despite a lack of correlation of the number or colony counts of EPCs, the ability of circulating progenitor cells to differentiate into an endothelial lineage is closely correlated with endothelial function. This cell function assay may serve as a novel biomarker for vascular function in healthy subjects in the pre-atherosclerotic stage.

Do stromal fibroblasts from basal cell carcinoma affect phenotype of normal keratinocytes? Epithelial-mesenchymal interactions are important not only to direct the course of prenatal development of skin and its appendages but also to influence the behaviour of transformed epithelial cells. Evaluation of the role of stromal fibroblasts on the phenotype of epithelial cells of basal cell carcinoma (BCC). The phenotype of human BCC was compared with the in vitro model where the growth and phenotypic pattern of normal human keratinocytes were monitored in co-culture with fibroblasts prepared from stroma of BCC (BCCFs), with normal dermal fibroblasts or with two established fibroblast lines. We visualized the expression of a panel of keratins, three types of endogenous lectin [galectin (Gal)-1, Gal-3 and Gal-7], binding sites for Gal-1 and Gal-3, a proliferation marker Ki67, nucleolar protein nucleostemin (NuclS) and membrane protein Ber-EP4. A phenotype and karyotype of BCCFs were also monitored. BCCFs were also grafted to NOD/LtSz-Rag1(null) mice to evaluate their malignant potential. Prolonged cultivation of BCCFs has led to morphological changes, loss of contact inhibition, loss of fibroblast surface antigens and progressive aneuploidity. However, a fully malignant phenotype did not develop as these cells did not form tumours in immunodeficient mice. Co-culture of BCCFs with normal keratinocytes in vitro led to their phenotypical changes resembling those in BCC, namely, expression of keratin 19. These keratinocytes also strongly express nuclear binding sites for Gal-1 and NuclS. This phenotype was not observed when normal keratinocytes were cultured with nontumour-originated fibroblasts.

These observations indicate that BCCFs may differ from normal fibroblasts and may play a regulatory role in BCC biology.

Is low-density lipoprotein receptor-related protein-associated protein ( LRPAP1 ) gene IVS5 insertion/deletion polymorphism a risk factor for gallstone disease in a Polish population? There is growing evidence that gallstone formation may be genetically determined. It was recently presented that a common polymorphism in the LRPAP1 gene might be associated with gallstone disease. Since reproducibility of data is important in genetic association studies, a case control study was designed to find out whether LRPAP1 gene polymorphism is associated with gallstone disease in a Polish population. Two hundred eighty-nine Polish Caucasian gallstone disease patients and 251 healthy controls participated in the study. A 37-bp insertion/deletion polymorphism in intron 5 of LRPAP1 (rs11267919) was determined by means of polymerase chain reaction assay. The frequencies and distribution of the insertion/deletion alleles did not differ significantly between gallstone disease patients and controls. No significant gender-related differences in allele frequencies or distributions were noted.

The LRPAP1 insertion/deletion polymorphism is not associated with gallstone disease in a Polish population.

Does vegetarian diet affect genes of oxidative metabolism and collagen synthesis? A vegetarian diet is known to prevent a series of diseases but may influence the balance of carbohydrate and fat metabolism as well as collagen synthesis. This study compares expression patterns of relevant genes in oral mucosa of omnivores and vegetarians. Quantitative reverse transcriptase polymerase chain reaction was applied for analysis of mRNA levels from carnitine transporter OCTN2, hepatic CPT1A and nonhepatic CPT1B isoforms of carnitine palmitoyltransferase and collagen (CCOL2A1) in oral mucosa. Compared with volunteers with traditional eating habits, carbohydrate consumption was significantly higher (+22%) in vegetarians. This was associated with a significant stimulation of CPT1A (+50%) and OCTN2 (+10%) and a lowered collagen synthesis (-10%).

These novel findings provide further insight into the association of a changed fat metabolism and reduced collagen synthesis in vegetarians, which could also play a role in the aging process.

Does continuous endotracheal tube cuff pressure control system protect against ventilator-associated pneumonia? The use of a system for continuous control of endotracheal tube cuff pressure reduced the incidence of ventilator-associated pneumonia (VAP) in one randomized controlled trial (RCT) with 112 patients but not in another RCT with 142 patients. In several guidelines on the prevention of VAP, the use of a system for continuous or intermittent control of endotracheal cuff pressure is not reviewed. The objective of this study was to compare the incidence of VAP in a large sample of patients (n = 284) treated with either continuous or intermittent control of endotracheal tube cuff pressure. We performed a prospective observational study of patients undergoing mechanical ventilation during more than 48 hours in an intensive care unit (ICU) using either continuous or intermittent endotracheal tube cuff pressure control. Multivariate logistic regression analysis (MLRA) and Cox proportional hazard regression analysis were used to predict VAP. The magnitude of the effect was expressed as odds ratio (OR) or hazard ratio (HR), respectively, and 95% confidence interval (CI). We found a lower incidence of VAP with the continuous (n = 150) than with the intermittent (n = 134) pressure control system (22.0% versus 11.2%; p = 0.02). MLRA showed that the continuous pressure control system (OR = 0.45; 95% CI = 0.22-0.89; p = 0.02) and the use of an endotracheal tube incorporating a lumen for subglottic secretion drainage (SSD) (OR = 0.39; 95% CI = 0.19-0.84; p = 0.02) were protective factors against VAP. Cox regression analysis showed that the continuous pressure control system (HR = 0.45; 95% CI = 0.24-0.84; p = 0.01) and the use of an endotracheal tube incorporating a lumen for SSD (HR = 0.29; 95% CI = 0.15-0.56; p < 0.001) were protective factors against VAP. However, the interaction between type of endotracheal cuff pressure control system (continuous or intermittent) and endotracheal tube (with or without SSD) was not statistically significant in MLRA (OR = 0.41; 95% CI = 0.07-2.37; p = 0.32) or in Cox analysis (HR = 0.35; 95% CI = 0.06-1.84; p = 0.21).

The use of a continuous endotracheal cuff pressure control system and/or an endotracheal tube with a lumen for SSD could help to prevent VAP in patients requiring more than 48 hours of mechanical ventilation.