Last-century forest productivity in a managed dry-edge Scots pine population: the two sides of climate warming.
Climate change in the Mediterranean, associated with warmer temperatures and more frequent droughts, is expected to impact forest productivity and the functioning of forests ecosystems as carbon reservoirs in the region. Climate warming can positively affect forest growth by extending the growing season, whereas increasing summer drought generally reduces forest productivity and may cause growth decline, trigger dieback, hamper regeneration, and increase mortality. Forest management could potentially counteract such negative effects by reducing stand density and thereby competition for water. The effectiveness of such interventions, however, has so far mostly been evaluated for short time periods at the tree and stand levels, which limits our confidence regarding the efficacy of thinning interventions over longer time scales under the complex interplay between climate, stand structure, and forest management. In this study, we use a century-long historical data set to assess the effects of climate and management on forest productivity. We consider rear-edge Scots pine (Pinus sylvestris) populations covering continental and Mediterranean conditions along an altitudinal gradient in Central Spain. We use linear mixed-effects models to disentangle the effects of altitude, climate, and stand volume on forest growth and ingrowth (recruitment and young trees' growth). We find that warming tends to benefit these tree populations, warmer winter temperature has a significant positive effect on both forest growth and ingrowth, and the effect is more pronounced at low elevations. However, drought conditions severely reduce growth and ingrowth, in particular when competition (stand volume) is high. We conclude that summer droughts are the main threat to Scots pine populations in the region, and that a reduction of stand volume can partially mitigate the negative impacts of more arid conditions. Mitigation and adaptation measures could therefore manage stand structure to adopt for the anticipated impacts of climate change in Mediterranean forest ecosystems.


Interrogating cardiac muscle cell mechanobiology on stiffness gradient hydrogels.
Extracellular matrix (ECM) remodeling is a major facet of cardiac development and disease, yet our understanding of cardiomyocyte mechanotransduction remains limited. To enhance our understanding of cardiomyocyte mechanosensation, we studied stiffness-driven changes to cell morphology and mechanomarker expression in H9C2 cells and neonatal rat cardiomyocytes (NRCMs). Linear stiffness gradient polyacrylamide hydrogels (2-33 kPa) coated with ECM proteins including Collagen I (Col), Fibronectin (Fn) or Laminin (Ln) were used to represent necrotic, healthy, and infarcted cardiac tissue on a continuous stiffness gradient. Cell size, cell shape and nuclear size were found to be mechanosensitive in H9C2 cells, as was the expression or nuclear translocalization of the mechanomarkers Lamin-A, YAP, and MRTF-A. Minor differences were observed between the different ECM coatings, with the same overarching stiffness-dependent trends being observed across Col, Fn and Ln coated hydrogels. Inhibition of mechanotransduction in H9C2 cells using blebbistatin or Y27632 resulted in disruptions to cell shape, nuclear shape, and nuclear size, however, trends in cell size and mechanomarker expression were not significantly attenuated. Mechanosensation in NRCMs was much less marked, with no significant changes in cell morphology being detected, although YAP did become increasingly nuclear localized with increasing stiffness. In α-actinin positive cells, striations formed with regular structure and frequency at all stiffnesses for Col and Fn coated hydrogels, but not Ln coated gels. In this study, we used our stiffness gradient hydrogels to comprehensively map the relationship between ECM stiffness and cardiac cell phenotype and found that less mature H9C2 cardiac cells are more sensitive to ECM changes than the more developed neonatal cardiomyocytes.


Holiday Discharges Are Associated with Higher 30-Day General Internal Medicine Hospital Readmissions at an Academic Medical Center.
Academic medical centers face unique challenges in educating physician trainees in effective discharge practices to prevent readmissions. Meanwhile, residents must handle high workloads coupled with frequent rotations to different services. This study aimed to determine whether daily service census, service turnover, time of discharge, and day of discharge increase the risk of 30-day readmission. All of the discharges from two academic general internal medicine teaching services between October 1, 2013 and September 30, 2014 were included in this observational data analysis. Variables were fit to a 30-day, all-cause readmission outcome using multiple logistic regression with inverse probability of treatment weighting and multiple imputations with chained equations. The following potential confounding variables were included in the model: health system utilization, demographics, laboratory values, and comorbidities. Among 1935 total discharges, 258 patients (13.3%) were readmitted within 30 days of the index discharge. Turnover, service census, weekend discharge, and time of discharge were not significantly associated with the risk of readmission. Patients discharged during holiday periods had higher odds of readmission (odds ratio 2.56, 95% confidence interval 2.01-3.25), whereas patients discharged on an intern switch day had lower odds of readmission (odds ratio 0.33, 95% confidence interval 0.27-0.41). Patients who are discharged during holiday periods are at a higher risk of readmission after adjusting for potential confounders. These results also suggest that discharge on an intern switch day had a protective effect on readmission. Further work is needed to examine whether these findings can be replicated, and, if confirmed, to determine to what extent these associations are causal.


Feasibility of a structured group education session to improve self-management of blood pressure in people with chronic kidney disease: an open randomised pilot trial.
Objectives We aimed to test, at pilot level, a structured group educational intervention to improve self-management of blood pressure in people with chronic kidney disease (CKD). The current paper explores patient acceptability of the intervention. Design This was an open randomised pilot trial. Participants were randomly assigned to either: A control group (n=41) receiving standard clinical management of hypertension. An intervention group (n=40) receiving standard clinical care plus the educational intervention. Setting Renal outpatient clinics at a single study centre. Participants Patients with early CKD and hypertension were identified and approached for recruitment. Intervention An evidence-based structured group educational intervention (CHEERS) using the principles of social cognitive theory to improve knowledge and self-management skills. Outcomes Recruitment, uptake of the intervention and patient satisfaction were evaluated to explore patient acceptability of the intervention and to determine any differences between patients regarding recruitment and retention. Measures Data on age, sex and ethnicity were collected for all patients approached to take part. For recruited patients, data were also collected on self-efficacy (ability to self-manage). Reasons given by patients declining to take part were recorded. Patients attending the educational session also completed an evaluation form to assess satisfaction. Results A total of 267 patients were approached, and 30% were randomly assigned. Lack of time (48%) and lack of interest (44%) were the main reasons cited for non-participation in the study. Men were significantly more likely to be recruited (p=0.048). The intervention was rated enjoyable and useful by 100% of participants. However, 37.5% of the intervention group failed to attend the educational session after recruitment. Participants failing to attend were significantly more likely to be older (p=0.039) and have lower self-efficacy (p=0.034). Conclusion The findings suggest that delivering and evaluating an effective structured group educational intervention to promote better blood pressure control in patients with CKD would be challenging in the current context of kidney care.


Genetic consequences of Pleistocene glaciations for the tundra vole (Microtus oeconomus) in Beringia.
Repeated glacial events during the Pleistocene fragmented and displaced populations throughout the northern continents. Different models of the effects of these climate-driven events predict distinct phylogeographic and population genetic outcomes for high-latitude faunas. The role of glaciations in (i) promoting intraspecific genetic differentiation and (ii) influencing genetic diversity was tested within a phylogeographic framework using the rodent Microtus oeconomus. The spatial focus for the study was Beringia, which spans eastern Siberia and northwestern North America, and was a continental crossroads and potential high arctic refugium during glaciations. Variation in mitochondrial DNA (cytochrome b and control region; 214 individuals) and nuclear DNA (ALDH1 intron; 63 individuals) was investigated across the Beringian region. Close genetic relationships among populations on either side of the Bering Strait are consistent with a history of periodic land connections between North America and Asia. A genetic discontinuity observed in western Beringia between members of a Central Asian clade and a Beringian clade is geographically congruent with glacial advances and with phylogeographic discontinuities identified in other organisms. Divergent island populations in southern Alaska were probably initially isolated by glacial vicariance, but subsequent differentiation has resulted from insularity. Tests of the genetic effects of postglacial colonization were largely consistent with expansion accompanied by founder effect bottlenecking, which yields reduced diversity in populations from recently deglaciated areas. Evidence that populations in the Beringian clade share a history of expansion from a low-diversity ancestral population suggests that Beringia was colonized by a small founder population from central Asia, which subsequently expanded in isolation.


Gender and nasal shape: measures for rhinoplasty.
Gender-specific nasal shapes are recommended for rhinoplasty. This study was conducted to clarify whether there truly are gender-related differences and to determine optimal nasal shapes in a Caucasian population. The authors created female and male composite photographs of "average" (n = 128 each), "optimal" (n = 16 each), and "most unpleasant" (n = 8 each) noses stratified on the basis of each photographed subject's (n = 311) own evaluation of the attractiveness of her or his nose, using a visual analogue scale. These composites were also assessed by 308 independent judges. Optimal female noses showed a horizontally and vertically lower nasion and were concave to straight in profile as compared with optimal male noses, which had a vertically and horizontally higher nasion and a straight profile. A supratip break was not found in any of the composites. At least half of the judges rated average and optimal male composite noses as female. A significant majority mistook the composite of the most unpleasant female noses as male (frontal view, 62.0 percent; lateral view, 72.4 percent; p < 0.001). Optimal and average female and male noses were found to be independently significantly more attractive than the most unpleasant ones (p < 0.001, n = 308 judges). Women and men with a straight or concave profile were significantly (p = 0.017 and p = 0.006, respectively) more satisfied with the appearance of their nose than those with nasal humps. Gender-related differences in nasal shape appear to be subtle, with nasion position being one of the main factors. A nasal hump and a supratip break are not desirable.


Phyllanthin from Phyllanthus amarus inhibits LPS-induced proinflammatory responses in U937 macrophages via downregulation of NF-κB/MAPK/PI3K-Akt signaling pathways.
Phyllanthin, a lignan from Phyllanthus species, has been reported to possess potent immunosuppressive properties on immune cells and on adaptive and innate immune responses in animal models. Herein, we investigated the inhibitory effects of phyllanthin isolated from Phyllanthus amarus on nuclear factor-kappa B (NF-κB), mitogen-activated protein kinase (MAPK), and PI3K-Akt signal transducing pathways in LPS-activated U937 cells. The lipopolysaccharide-stimulated excess production of prostaglandin was significantly suppressed by phyllanthin via the mechanisms linked to the modulatory effects of cyclooxygenase 2 protein and gene expression. Phyllanthin also significantly inhibited the release and mRNA expression of proinflammatory cytokines (interleukin-1 beta and tumor necrosis factor-alpha). Phyllanthin also significantly downregulated the phosphorylation of IκBα, NF-κB (p65), and IKKα/β and suppressed the activation of JNK, ERK, p38MAPK, and Akt in a concentration-dependent manner. Additionally, phyllanthin downregulated the expression of upstream signaling molecules including MyD88 and toll-like receptor 4 that are essential for the activation of NF-κB, MAPKs, and PI3K-Akt signal transducing pathways. Based on these observations, phyllanthin may exert their suppressive effects on inflammatory process by mediating the release of inflammatory signaling molecules via the NF-κB, MAPKs, and PI3K-Akt signal transducing pathways. Thus, phyllanthin holds a great promise as a potential anti-inflammatory agent to treat various inflammatory diseases.


Environmental origins of sexually selected variation and a critique of the fluctuating asymmetry-sexual selection hypothesis.
Identifying sources of phenotypic variability in secondary sexual traits is critical for understanding their signaling properties, role in sexual selection, and for predicting their evolutionary dynamics. The present study tests for the effects of genotype, developmental temperature, and their interaction, on size and fluctuating asymmetry of the male sex comb, a secondary sexual character, in Drosophila bipectinata Duda. Both the size and symmetry of elements of the sex comb have been shown previously to be under sexual selection in a natural population in northeastern Australia. Two independent reciprocal crosses were conducted at 25 degrees and 29 degrees C between genetic lines extracted from this population that differed in the size of the first (TC1) and third (TC3) comb segments. These temperatures are within the documented range experienced by the species in nature. Additive and dominance genetic effects were detected for TC1, whereas additive genetic, and Y-chromosomal effects were detected for TC3. TC2 and TC3 decreased sharply with increasing temperature, by 10% and 22%, respectively. In contrast, positional fluctuating asymmetry (PFA) significantly increased with temperature, by up to 38%. The results (1) document an important source of environmental variance in a sexual ornament expected to reduce trait heritability in field populations, and thus act to attenuate response to sexual selection, (2) suggest that variation in ornament size reflects differences in male condition; and (3) support the general hypothesis that asymmetry in a sexual ornament is indicative of developmental instability arising from environmental stress. The "environmental heterogeneity" (EH) hypothesis is proposed, and supportive evidence for it presented, to explain negative size-FA correlations in natural populations. Data and theory challenge the use of negative size-FA correlations observed in nature to support the FA-sexual selection hypothesis, which posits that such correlations are driven by differences in genetic quality among individuals.


Calsequestrin and the calcium release channel of skeletal and cardiac muscle.
Calsequestrin is by far the most abundant Ca(2+)-binding protein in the sarcoplasmic reticulum (SR) of skeletal and cardiac muscle. It allows the Ca2+ required for contraction to be stored at total concentrations of up to 20mM, while the free Ca2+ concentration remains at approximately 1mM. This storage capacity confers upon muscle the ability to contract frequently with minimal run-down in tension. Calsequestrin is highly acidic, containing up to 50 Ca(2+)-binding sites, which are formed simply by clustering of two or more acidic residues. The Kd for Ca2+ binding is between 1 and 100 microM, depending on the isoform, species and the presence of other cations. Calsequestrin monomers have a molecular mass of approximately 40 kDa and contain approximately 400 residues. The monomer contains three domains each with a compact alpha-helical/beta-sheet thioredoxin fold which is stable in the presence of Ca2+. The protein polymerises when Ca2+ concentrations approach 1mM. The polymer is anchored at one end to ryanodine receptor (RyR) Ca2+ release channels either via the intrinsic membrane proteins triadin and junctin or by binding directly to the RyR. It is becoming clear that calsequestrin has several functions in the lumen of the SR in addition to its well-recognised role as a Ca2+ buffer. Firstly, it is a luminal regulator of RyR activity. When triadin and junctin are present, calsequestrin maximally inhibits the Ca2+ release channel when the free Ca2+ concentration in the SR lumen is 1mM. The inhibition is relieved when the Ca2+ concentration alters, either because of small changes in the conformation of calsequestrin or its dissociation from the junctional face membrane. These changes in calsequestrin's association with the RyR amplify the direct effects of luminal Ca2+ concentration on RyR activity. In addition, calsequestrin activates purified RyRs lacking triadin and junctin. Further roles for calsequestrin are indicated by the kinase activity of the protein, its thioredoxin-like structure and its influence over store operated Ca2+ entry. Clearly, calsequestrin plays a major role in calcium homeostasis that extends well beyond its ability to buffer Ca2+ ions.


Rotation of Four Small Nitroxide Probes in Supercooled Bulk Water.
Using a precise method of least-squares nonlinear electron paramagnetic resonance (EPR) line fitting, we have obtained experimental evidence of a decoupling of the rotational motion of four nitroxide spin probes from the viscosity of bulk water at 277 K. This decoupling is about 50 K higher than another such phenomenon observed in interstitial supercooled water of polycrystalline ice by Banerjee et al. (Proc Natl Acad Sci USA 106 (2009) 11448-11453). Above 277 K the activation energies of the rotation of the probes and water viscosity are very close, while in the supercooled region the activation energies of the probes' rotation are greater than that of the viscosity of water. The rotational correlation times of the probes can be fit well to a power law functionality with a singular temperature. The temperature dependence of the hydrodynamic radii of the probes indicates two distinct dynamical regions, which cross at 277 K.


Contractile effects of the exchange of cardiac troponin for fast skeletal troponin in rabbit psoas single myofibrils.
The effects of the removal of fast skeletal troponin C (fsTnC) and its replacement by cardiac troponin C (cTnC) and the exchange of fast skeletal troponin (fsTn) for cardiac troponin (cTn) were measured in rabbit fast skeletal myofibrils. Electrophoretic analysis of myofibril suspensions indicated that replacement of fsTnC or exchange of fsTn with cTnC or cTn was about 90% complete in the protocols used. Mechanical measurements in single myofibrils, which were maximally activated by fast solution switching, showed that replacement of fsTnC with cTnC reduced the isometric tension, the rate of tension rise following a step increase in Ca2+ (kACT), and the rate of tension redevelopment following a quick release and restretch (kTR), but had no effect on the kinetics of the fall in tension when the concentration of inorganic phosphate (Pi) was abruptly increased (kPi(+)). These data suggest that the chimeric protein produced by cTnC replacement in fsTn alters those steps controlling the weak-to-strong crossbridge attachment transition. Inefficient signalling within the chimeric troponin may cause these changes. However, replacement of fsTn by cTn had no effect on maximal isometric tension, kACT or kTR, suggesting that these mechanics are largely determined by the isoform of the myosin molecule. Replacement of fsTn by cTn, on the other hand, shifted the pCa50 of the pCa-tension relationship from 5.70 to 6.44 and reduced the Hill coefficient from 3.3 to 1.4, suggesting that regulatory protein isoforms primarily alter Ca2+ sensitivity and the cooperativity of the force-generating mechanism.


The application of the parallel track model in community health promotion: a literature review.
The parallel track model is one of the several models that are used in health promotion programmes that focus on community empowerment. It is unique in that it explicitly incorporates an empowerment approach with a top-down health programme. Since its development in 1999-2000 the model has been used in various health programmes in both developed and developing countries. The aim of this review is to examine the nature and extent of the application of this model and its contribution to promoting health. A review of the literature published between 2000 and 2011 was conducted. Nine results matched the inclusion criteria and revealed that the model has been mostly applied to disadvantaged communities to address health determinants, such as poverty and health literacy. This review found that the model had a positive impact on specific health outcomes such as health literacy and community capacity. We concluded that the parallel track model has the most potential for building capacity for community health promotion and appears to be the least useful for interventions focusing on health behaviour change within a limited time frame.


[Stages of kinetotherapy and therapeutic massage for long time bedridden patients].
Rehabilitation of stroke patients, is a complex chain of consistently conducted medical and social measures aimed at rehabilitation, health or possible adaptation to self-service in the presence of a persistent neurological defect. Already in acute stroke patient needs not only to drug therapy and care, but also in the emotional and psychological support for the type of "cautious optimism." It should be explained to the patient a temporary, reversible nature of its existing motor and other violations. With relatives patient should discuss the real situation, prognosis, rehabilitation opportunities, the need for hospitalization or appropriate treatment at home, put them in front of clear objectives for the near future. Activity and the amount of rehabilitation expanded in young children with satisfactory prognosis, consideration being given to their professional and social status.


A prospective study examining cachexia predictors in patients with incurable cancer.
Early intervention against cachexia necessitates a predictive model. The aims of this study were to identify predictors of cachexia development and to create and evaluate accuracy of a predictive model based on these predictors. A secondary analysis of a prospective, observational, multicentre study was conducted. Patients, who attended a palliative care programme, had incurable cancer and did not have cachexia at baseline, were amenable to the analysis. Cachexia was defined as weight loss (WL) > 5% (6 months) or WL > 2% and body mass index< 20 kg/m2. Clinical and demographic markers were evaluated as possible predictors with Cox analysis. A classification and regression tree analysis was used to create a model based on optimal combinations and cut-offs of significant predictors for cachexia development, and accuracy was evaluated with a calibration plot, Harrell's c-statistic and receiver operating characteristic curve analysis. Six-hundred-twenty-eight patients were included in the analysis. Median age was 65 years (IQR 17), 359(57%) were female and median Karnofsky performance status was 70(IQR 10). Median follow-up was 109 days (IQR 108), and 159 (25%) patients developed cachexia. Initial WL, cancer type, appetite and chronic obstructive pulmonary disease were significant predictors (p ≤ 0.04). A five-level model was created with each level carrying an increasing risk of cachexia development. For Risk-level 1-patients (WL < 3%, breast or hematologic cancer and no or little appetite loss), median time to cachexia development was not reached, while Risk-level 5-patients (WL 3-5%) had a median time to cachexia development of 51 days. Accuracy of cachexia predictions at 3 months was 76%. Important predictors of cachexia have been identified and used to construct a predictive model of cancer cachexia. ClinicalTrials.gov Identifier: NCT01362816 .


[Diagnosis of healthcare issues in clinics and hospital of Barranquilla].
Objective To identify and assess the main healthcare issues found in clinics and hospitals in Barranquilla. Methods Descriptive study applied on two populations: clinics [23] and hospitals [5]. A confidence level of 95 % and the alpha level of 5 % and p=0.5 were used in the study. The resulting sample size for clinics and hospitals was 18 and 4, respectively. Clinics and hospitals were randomly and a 21-question survey was designed to find out the status of the different healthcare processes in the Health Care Sector. The results were processed by using Microsoft Excel 2010 software. Results On one hand, 50 % of the hospitals expressed having problems in outpatient, hospitalization and statistical departments. On the other hand, 61.1 % of the clinics have difficulties in Emergency rooms, 50 % in Surgical Services, 50% in Hospitalization and 38.9 % in Outpatient Department. Conclusions The diagnosis regarding healthcare issues in clinics and hospitals of Barranquilla determines that although the Hospitalization process is a common point for potential improvement in both hospitals and clinics of the city, the greatest priority should be given to Surgical Services, Emergency Department and Statistical Department, due to their average intervention priority.


Association of the KIAA0319 dyslexia susceptibility gene with reading skills in the general population.
The authors previously identified a haplotype on chromosome 6p22 defined by three single-nucleotide polymorphisms (SNPs) that was associated with dyslexia (reading disability) in two independent samples of families that included at least one sibling with severe reading impairment. The authors also showed that this haplotype is associated with a reduction in expression of the KIAA0319 gene. In addition, a completely independent study detected an association between KIAA0319 markers and reading disability. In the current study, the authors tested whether the KIAA0319 gene influences reading skills in the general population, rather than having an effect restricted to reading disability. The authors genotyped four SNPs that previously showed association with reading disability in the population of 7-9-year-old children in the Avon Longitudinal Study of Parents and Children (ALSPAC), a large longitudinal cohort for which reading-related phenotypes were available for more than 6,000 individuals. The authors conducted quantitative analysis for both single markers and haplotypes. The rs2143340 SNP, which effectively tags the three-SNP risk haplotype, was significantly associated with a test for reading ability. The risk haplotype itself also showed association with poor reading performance, and as in previous research, the association was stronger when the analysis was controlled for IQ. These results both support a role of the KIAA0319 gene in the development of dyslexia and suggest that this gene influences reading ability in the general population. Moreover, the data implicate the three-SNP haplotype and its tagging SNP rs2143340 as genetic risk factors for poor reading performance.


N-Acetylcysteine Compared to Metformin, Improves The Expression Profile of Growth Differentiation Factor-9 and Receptor Tyrosine Kinase c-Kit in The Oocytes of Patients with Polycystic Ovarian Syndrome.
Paracrine disruption of growth factors in women with polycystic ovarian syndrome (PCOS) results in production of low quality oocyte, especially following ovulation induction. The aim of this study was to investigate the effects of metformin (MET), N-acetylcysteine (NAC) and their combination on the hormonal levels and expression profile of GDF-9, BMP-15 and c-kit, as hallmarks of oocyte quality, in PCOS patients. This prospective randomized, double-blind, placebo controlled trial aims to study the effects of MET, NAC and their combination (MET+NAC) on expression of GDF-9, BMP-15 and c-kit mRNA in oocytes [10 at the germinal vesicle (GV) stage, 10 at the MI stage, and 10 at the MII stage from per group] derived following ovulation induction in PCOS. Treatment was carried out for six weeks, starting on the third day of previous cycle until oocyte aspiration. The expression of GDF9, BMP15 and c-kit were determined by quantitative real time polymerase chain reaction (RT-qPCR) and western blot analysis. Data were analyzed with one-way ANOVA. The follicular fluid (FF) level of c-kit protein significantly decreased in the NAC group compared to the other groups. Significant correlations were observed between the FF soluble c-kit protein with FF volume, androstenedione and estradiol. The GDF-9 expression in unfertilized mature oocytes were significantly higher in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups. The c-kit expression in unfertilized mature oocytes were significantly lower in the NAC group compared to the other groups (P<0.001). Similar difference was not observed between the MET, NAC+MET and control groups (Registration number: IRCT201204159476N1). We concluded that NAC can improve the quality of oocytes in PCOS.


HGF regulates the development of cortical pyramidal dendrites.
Although hepatocyte growth factor (HGF) and its receptor tyrosine kinase MET are widely expressed in the developing and mature central nervous system, little is known about the role of MET signaling in the brain. We have used particle-mediated gene transfer in cortical organotypic slice cultures established from early postnatal mice to study the effects of HGF on the development of dendritic arbors of pyramidal neurons. Compared with untreated control cultures, exogenous HGF promoted a highly significant increase in dendritic growth and branching of layer 2 pyramidal neurons, whereas inactivation of endogenous HGF with function-blocking, anti-HGF antibody caused a marked reduction in size and complexity of the dendritic arbors of these neurons. Furthermore, pyramidal neurons transfected with an MET dominant-negative mutant receptor likewise had much smaller and less complex dendritic arbors than did control transfected neurons. Our results indicate that HGF plays a role in regulating dendritic morphology in the developing cerebral cortex.


Evaluation of three formulations of Beauveria bassiana for control of lesser mealworm and hide beetle in Georgia poultry houses.
Initial screening of 12 Beauveria bassiana (Balsamo) Vuillemin isolates against larvae of the lesser mealworm (Alphitobius diaperinus [Panzer]) resulted in the selection of two isolates, GHA and 707, for further testing under field conditions. Three formulations of each strain were prepared: an EC, a ground corn granular formulation, and waste product of fungal propagation containing spent media, mycelia, and unharvested conidia ("residue" formulation). Two field trials were conducted in commercial caged-layer houses in Georgia with 5-6 mo of manure accumulation and established populations of A. diaperinus and hide beetles (Dermestes maculatus DeGeer). In the first trial field, B. bassiana was applied a single time to the manure surface at either 10(9) (EC and granular cornmeal bait formulations) or 10(8) (residue formulation) fungal spores per square meter. In the second trial, two successive weekly treatments were applied, using a total of 6x the rate of application used in the first trial, Significant treatment effects were short-lived and only detected 2 wk after treatment in both trials. The granular formulations of both strains and the residue formulation of the GHA strain provided the greatest degree of suppression (60-90%) of beetle larvae. A laboratory bioassay confirmed that the granular bait was the most effective formulation. More frequent applications made earlier in the manure accumulation cycle may be necessary to achieve satisfactory control of these beetles.


The dermal sensitization threshold (DST) approach for mixtures evaluated as negative in in vitro test methods; mixture DST.
Cosmetic ingredients often comprise complex mixtures, such as botanical extracts, which may contain skin sensitizing constituents. In our previous study for the sensitivity of the evaluations of skin sensitizing constituents in mixtures using the binary in vitro test battery with KeratinoSensTM and h-CLAT, some sensitizers showed higher detection limits in in vitro test methods than in murine local lymph node assays (LLNA). Thus, to minimize the uncertainty associated with decreased sensitivity for these sensitizers, a risk assessment strategy was developed for mixtures with negative results from the binary test battery. Assuming that the no expected sensitization induction level of mixtures (mixture NESIL) can be derived for mixtures with negative in vitro test results, we assessed 146 sensitizers with in vitro and LLNA data according to the assumption of indeterminate constituents in mixtures. Finally, we calculated 95th percentile probabilities of mixture NESILs and derived dermal sensitization thresholds for mixtures (mixture DST) with negative in vitro test results of 6010 μg/cm2. Feasibility studies indicated that this approach was practical for risk assessments of products in the cosmetic industry. This approach would be a novel risk assessment strategy for incorporating the DST approach and information from in vitro test methods.


Inverse relationship between six week postvaccination varicella antibody response to vaccine and likelihood of long term breakthrough infection.
We used the large clinical database that supported the development of Oka/ Merck varicella vaccine to study the relationship between the primary varicella antibody response, as determined by gpELISA, an enzyme-linked immunosorbent assay that detects antibodies to varicella-zoster virus (VZV) glycoprotein, and the subsequent risk of postvaccination breakthrough varicella. We vaccinated 1,164 healthy children with a single dose of varicella vaccine containing 2900 to 9000 plaque-forming units/dose. The primary immune response to vaccination was determined by gpELISA 6 weeks after vaccination. Subjects were followed annually for 7 years to ascertain cases of breakthrough varicella. The estimated vaccine efficacy among children with a 6-week postvaccination antibody titer of > or = 5 gpELISA units was 95.5% (95% confidence interval, 94.2%, 96.8%) compared with 83.5% (95% confidence interval, 76.9%, 89.5%) for subjects with a titer of <5 gpELISA units. Children with a 6-week postvaccination antibody titer of <5 gpELISA units were 3.5 times more likely than those with a titer of > or = 5 gpELISA units to develop breakthrough varicella. We identified a 6-week postvaccination antibody titer of > or = 5 gpELISA units as an approximate correlate of protection. In addition we established an accelerated failure time model based on log normal hazard that predicted varicella breakthrough rates based on the distribution of 6-week postvaccination varicella antibody titers.


Visual Attention Patterns Differ in Gynephilic and Androphilic Men and Women Depending on Age and Gender of Targets.
Visual attention patterns measured with eye-tracking techniques provide indirect clues about sexual response. This study aimed to test the category specificity of sexual responses to stimuli varying in gender and age by evaluating both early and late attention of gynephilic and androphilic men and women. We simultaneously presented sexually preferred and nonpreferred stimuli and measured time to first fixation and total duration of fixation on four areas of interest: entire body, then face, chest, and pelvis. Androphilic women's early attention patterns were nonspecific, whereas gynephilic women and both groups of men showed a category-specific pattern for the entire body. In contrast, all groups showed gender-specific patterns of late attention for all areas of interest. We also found support for age specificity of early and late visual attention in all four groups, with greater attention to adult than child stimuli. This study supports the usefulness of a competing stimulus eye-tracking paradigm as a method to examine gender specificity in gynephilic women and androphilic and gynephilic men, and as a measure of age specificity in gynephilic and androphilic men and women.


Transcriptomic analysis of Arabidopsis thaliana plants treated with the Ky-9 and Ky-72 histone deacetylase inhibitors.
Histone acetylation plays a pivotal role in plant growth and development, and is regulated by the antagonistic relationship between histone acetyltransferase (HAT) and histone deacetylase (HDAC). We previously revealed that some HDAC inhibitors confer high-salinity stress tolerance in plants. In this study, we identified two HDAC inhibitors, namely Ky-9 and Ky-72, which enhanced the high-salinity stress tolerance of Arabidopsis thaliana. Ky-9 and Ky-72 are structurally similar chlamydocin analogs. However, the in vitro inhibitory activity of Ky-9 against mammalian HDAC is greater than that of Ky-72. A western blot indicated that Ky-9 and Ky-72 increased the acetylation levels of histone H3, suggesting they exhibit HDAC inhibitory activities in plants. We conducted a transcriptomic analysis to investigate how Ky-9 and Ky-72 enhance high-salinity stress tolerance. Although Ky-9 upregulated the expression of more genes than Ky-72, similar gene expression patterns were induced by both HDAC inhibitors. Additionally, the expression of high-salinity stress tolerance-related genes, such as anthocyanin-related genes and a small peptide-encoding gene, increased by Ky-9 and Ky-72. These data suggest that slight structural differences in chemical side chain between HDAC inhibitors can alter inhibitory effect on HDAC protein leading to influence gene expression, thereby enhancing high-salinity stress tolerance in different extent.


Mutations in the gene encoding starch synthase II profoundly alter amylopectin structure in pea embryos.
Mutations at the rug5 (rugosus5) locus have been used to elucidate the role of the major soluble isoform of starch synthase II (SSII) in amylopectin synthesis in the developing pea embryo. The SSII gene maps to the rug5 locus, and the gene in one of three rug5 mutant lines has been shown to carry a base pair substitution that introduces a stop codon into the open reading frame. All three mutant alleles cause a dramatic reduction or loss of the SSII protein. The mutations have pleiotropic effects on the activities of other isoforms of starch synthase but apparently not on those of other enzymes of starch synthesis. These mutations result in abnormal starch granule morphology and amylopectin structure. Amylopectin contains fewer chains of intermediate length (B2 and B3 chains) and more very short and very long chains than does amylopectin from wild-type embryos. The results suggest that SSII may play a specific role in the synthesis of B2 and B3 chains of amylopectin. The extent to which these findings can be extrapolated to other species is discussed.


Somatosensory thalamic neurons: effects of cortical depression.
Deafferented somatosensory thalamic neurons showed hyperactivity, followed by greatly reduced activity, after initiation of cortical spreading depression; local cooling of sensorimotor cortex was followed only by the inactive phase. Stimulation of contralateral midbrain reticular formation during the inactive phase failed to induce the typical increase in discharge rate of somatosensory thalamic neurons, but produced desynchronization in unaffected cortex. These results indicate that corticothalamic discharge is necessary for sustaining the ongoing activity of deafferented somatosensory thalamic neurons and for maintaining their responsiveness to stimulation of the reticular formation.


The genome-wide dynamics of the binding of Ldb1 complexes during erythroid differentiation.
One of the complexes formed by the hematopoietic transcription factor Gata1 is a complex with the Ldb1 (LIM domain-binding protein 1) and Tal1 proteins. It is known to be important for the development and differentiation of the erythroid cell lineage and is thought to be implicated in long-range interactions. Here, the dynamics of the composition of the complex-in particular, the binding of the negative regulators Eto2 and Mtgr1-are studied, in the context of their genome-wide targets. This shows that the complex acts almost exclusively as an activator, binding a very specific combination of sequences, with a positioning relative to transcription start site, depending on the type of the core promoter. The activation is accompanied by a net decrease in the relative binding of Eto2 and Mtgr1. A Chromosome Conformation Capture sequencing (3C-seq) assay also shows that the binding of the Ldb1 complex marks genomic interaction sites in vivo. This establishes the Ldb1 complex as a positive regulator of the final steps of erythroid differentiation that acts through the shedding of negative regulators and the active interaction between regulatory sequences.


Automation in clinical biochemistry: core, peripheral, STAT, and specialist laboratories in Australia.
Pathology has developed substantially since the 1990s with the introduction of total laboratory automation (TLA), in response to workloads and the need to improve quality. TLA has enhanced core laboratories, which evolved from discipline-based laboratories. Work practices have changed, with central reception now loading samples onto the Inlet module of the TLA. It is important to continually appraise technology. This study looked at the impact of technology using a self-administered survey to seniors in clinical biochemistry in NATA GX/GY-classified laboratories in Australia. The responses were yes, no, or not applicable and are expressed as percentages of responses. Some of the questions sourced for descriptive answers. Eighty-one laboratories responded, and the locations were 63%, 33%, and 4% in capital cities, regional cities, and country towns, respectively. Forty-two percent were public and 58% private. Clinical biochemistry was in all core laboratories of various sizes, and most performed up to 20 tests per sample. Thirty percent of the 121 surveyed laboratories had plans to install an automated line. Fifty-eight percent had hematology and biochemistry instrumentations in their peripheral laboratory, and 16% had a STAT laboratory on the same site as the core laboratory. There were varied instruments in specialist laboratories, and analyzers with embedded computers were in all laboratories. Medium and large laboratories had workstations with integrated instruments, and some large laboratories had TLA. Technology evolution and rising demand for pathology services make it imperative for laboratories to embrace such changes and reorganize the laboratories to take into account point-of-care testing and the efficiencies of core laboratories and TLA.


The Impact of Biomaterial Cell Contact on the Immunopeptidome.
Biomaterials play an increasing role in clinical applications and regenerative medicine. A perfectly designed biomaterial should restore the function of damaged tissue without triggering an undesirable immune response, initiate self-regeneration of the surrounding tissue and gradually degrade after implantation. The immune system is well recognized to play a major role in influencing the biocompatibility of implanted medical devices. To obtain a better understanding of the effects of biomaterials on the immune response, we have developed a highly sensitive novel test system capable of examining changes in the immune system by biomaterial. Here, we evaluated for the first time the immunopeptidome, a highly sensitive system that reflects cancer transformation, virus or drug influences and passes these cellular changes directly to T cells, as a test system to examine the effects of contact with materials. Since monocytes are one of the first immune cells reacting to biomaterials, we have tested the influence of different materials on the immunopeptidome of the monocytic THP-1 cell line. The tested materials included stainless steel, aluminum, zinc, high-density polyethylene, polyurethane films containing zinc diethyldithiocarbamate, copper, and zinc sulfate. The incubation with all material types resulted in significantly modulated peptides in the immunopeptidome, which were material-associated. The magnitude of induced changes in the immunopeptidome after the stimulation appeared comparable to that of bacterial lipopolysaccharides (LPS). The source proteins of many detected peptides are associated with cytotoxicity, fibrosis, autoimmunity, inflammation, and cellular stress. Considering all tested materials, it was found that the LPS-induced cytotoxicity-, inflammation- and cellular stress-associated HLA class I peptides were mainly induced by aluminum, whereas HLA class II peptides were mainly induced by stainless steel. These findings provide the first insights into the effects of biomaterials on the immunopeptidome. A more thorough understanding of these effects may enable the design of more biocompatible implant materials using in vitro models in future. Such efforts will provide a deeper understanding of possible immune responses induced by biomaterials such as fibrosis, inflammation, cytotoxicity, and autoimmune reactions.


Thermodynamics of zinc binding to hepatitis C virus NS3 protease: a folding by binding event.
The hepatitis C virus (HCV) nonstructural protein 3 (NS3) protease is responsible for the processing of the non-structural region of the viral precursor polyprotein in infected hepatic cells. HCV NS3 is a zinc-dependent serine protease. The zinc ion, which is bound far away from the active site and considered to have a structural role, is essential for the structural integrity of the protein; furthermore, the ion is required for the hydrolytic activity. Consequently, the NS3 zinc binding site has been considered for a long time as a possible target for drug discovery. As a first step towards this goal, the energetics of the NS3-zinc interaction and its effect on the NS3 conformation must be established and discussed. The thermodynamic characterization of zinc binding to NS3 protease by isothermal titration calorimetry and spectroscopy is presented here. Spectroscopic and calorimetric results suggest that a considerable conformational change in the protein is coupled to zinc binding. The energetics of the conformational change is comparable to that of the folding of a protein of similar size. Therefore, zinc binding to NS3 protease can be considered as a "folding by binding" event.


Domain characterization of rabbit skeletal muscle myosin light chain kinase.
Myosin light chain kinase can be divided into three distinct structural domains, an amino-terminal "tail," of unknown function, a central catalytic core and a carboxy-terminal calmodulin-binding regulatory region. We have used a combination of deletion mutagenesis and monoclonal antibody epitope mapping to define these domains more closely. A 2.95-kilobase cDNA has been isolated that includes the entire coding sequence of rabbit skeletal muscle myosin light chain kinase (607 amino acids). This cDNA, expressed in COS cells encoded a Ca2+/calmodulin-dependent myosin light chain kinase with a specific activity similar to that of the enzyme purified from rabbit skeletal muscle. Serial carboxy-terminal deletions of the regulatory and catalytic domains were constructed and expressed in COS cells. The truncated kinases had no detectable myosin light chain kinase activity. Monoclonal antibodies which inhibit the activity of the enzyme competitively with respect to myosin light chain were found to bind between residues 235-319 and 165-173, amino-terminal of the previously defined catalytic core. Thus, residues that are either involved in substrate binding or in close proximity to a light chain binding site may be located more amino-terminal than the previously defined catalytic core.


Adaptation in the asexual false spider mite Brevipalpus phoenicis: evidence for frozen niche variation.
Because asexual species lack recombination, they have little opportunity to produce genetically variable offspring and cannot adapt to changes in their environment. However, a number of asexual species are very successful and appear to contradict this general view. One such species is the phytophagous mite Brevipalpus phoenicis (Geijskes), a species that is found in a wide range of environments. There are two general explanations for this pattern, the General Purpose Genotype (GPG) and Frozen Niche Variation (FNV). According to the GPG model, an asexual species consists of clones that can all survive and reproduce in all the different niches. Alternatively, the FNV model postulates that different clones are specialized to different niches. We have performed a test to distinguish between these models in B. phoenicis. Mites from three populations from three different host plant species (citrus, hibiscus and acerola) were transplanted to their own and the two alternative host plants and mite survival and egg production were measured. Additionally, the mite populations were genotyped using microsatellites. Fitness was seriously reduced when mites were transplanted to the alternative host plant species, except when the alternative host was acerola. We concluded that B. phoenicis clones are specialized to different niches and thus the FNV best describes the broad ecological niche of this species but that there is also some evidence for host plant generalization. This conclusion was strengthened by the observations that on each host plant species the native mite population performed better than the introduced ones, and that three microsatellite markers showed that the mite populations are genetically distinct.


Anaerobic induction of the alkylation-inducible Escherichia coli aidB gene involves genes of the cysteine biosynthetic pathway.
The Escherichia coli aidB gene is a component of the adaptive response to alkylation damage. This gene is subject to two different forms of induction: an ada-dependent alkylation induction and an ada-independent induction that occurs when cells are grown anaerobically (M. R. Volkert, L. I. Hajec, and D. C. Nguyen, J. Bacteriol. 171:1196-1198, 1989; M. R. Volkert, and D. C. Nguyen, Proc. Natl. Acad. Sci. USA 81:4110-4114, 1984). In this study, we isolated and characterized strains bearing mutations that specifically affect the anaerobic induction pathway. This pathway requires a functional cysA operon, which encodes sulfate permease. Mutations in cysA block this pathway of aidB induction. In contrast, mutations in either cysH, cysD, cysN, or cysC result in elevated levels of aidB expression during aerobic growth. These results indicate that the sulfate transport genes perform a role in anaerobic induction of the aidB gene and suggest that growth under anaerobic conditions may modify either the function or the expression of gene products encoded by the cysA operon.


Optimal binary solvent extraction system for phenolic antioxidants from mengkudu (Morinda citrifolia) fruit.
Antioxidants have been widely used in the food industry to enhance product quality by preventing oxidation of susceptible substances. This work was carried out to maximise the recovery of total phenolic content (TPC), total flavonoid content (TFC), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical-scavenging capacity and 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging capacity from Morinda citrifolia fruit via modification of the ethanol concentration, extraction time and extraction temperature at minimal processing cost. The optimised conditions yielded values of 881.57 ± 17.74 mg GAE/100 g DW for TPC, 552.53 ± 34.16 mg CE/100 g DW for TFC, 799.20 ± 2.97 µmol TEAC/100 g DW for ABTS and 2,317.01 ± 18.13 µmol TEAC/100 g DW for DPPH were 75% ethanol, 40 min of time and 57 °C. The four responses did not differ significantly (p > 0.05) from predicted values, indicating that models obtained are suitable to the optimisation of extraction conditions for phenolics from M. citrifolia. The relative amounts of flavonoids were 0.784 ± 0.01 mg quercetin/g of extract and 1.021 ± 0.04 mg rutin/g of extract. On the basis of the results obtained, M. citrifolia extract can be used as a valuable bioactive source of natural antioxidants.


Epidemiology of vancomycin-resistant enterococci in the United Arab Emirates: a retrospective analysis of 12 years of national AMR surveillance data.
Enterococci are usually low pathogenic, but can cause invasive disease under certain circumstances, including urinary tract infections, bacteremia, endocarditis, and meningitis, and are associated with peritonitis and intra-abdominal abscesses. Increasing resistance of enterococci to glycopeptides and fluoroquinolones, and high-level resistance to aminoglycosides is a concern. National antimicrobial resistance (AMR) surveillance data for enterococci from the Middle East and North Africa (MENA) and the Gulf region is scarce. A retrospective 12-year analysis of N = 37,909 non-duplicate diagnostic Enterococcus spp. isolates from the United Arab Emirates (UAE) was conducted. Data was generated by routine patient care during 2010-2021, collected by trained personnel and reported by participating surveillance sites to the UAE National AMR Surveillance program. Data analysis was conducted with WHONET. Enterococcus faecalis was the most commonly reported species (81.5%), followed by Enterococcus faecium (8.5%), and other enterococci species (4.8%). Phenotypically vancomycin-resistant enterococci (VRE) were found in 1.8% of Enterococcus spp. isolates. Prevalence of VRE (%VRE) was highest for E. faecium (8.1%), followed by E. faecalis (0.9%). A significant level of resistance to glycopeptides (%VRE) for these two species has been observed in the majority of observed years [E. faecalis (0-2.2%), 2010: 0%, 2021: 0.6%] and E. faecium (0-14.2%, 2010: 0%, 2021: 5.8%). Resistance to fluoroquinolones was between 17 and 29% (E. faecalis) and was higher for E. faecium (between 42 and 83%). VRE were associated with higher patient mortality (RR: 2.97), admission to intensive care units (RR: 2.25), and increased length of stay (six excess inpatient days per VRE case), as compared to vancomycin-susceptible Enterococcus spp. Published data on Enterococcus infections, in particular VRE-infections, in the UAE and MENA region is scarce. Our data demonstrates that VRE-enterococci are relatively rare in the UAE, however showing an increasing resistance trend for several clinically important antibiotic classes, causing a concern for the treatment of serious infections caused by enterococci. This study also demonstrates that VRE were associated with higher mortality, increased intensive care unit admission rates, and longer hospitalization, thus poorer clinical outcome and higher associated costs in the UAE. We recommend the expansion of current surveillance techniques (e.g., local VRE screening), stricter infection prevention and control strategies, and better stewardship interventions. Further studies on the molecular epidemiology of enterococci are needed.


Assessment of optimal renin-angiotensin-system inhibition strategy in Asian patients with STEMI after primary myocardial revascularization.
For the Asian patients with STEMI undergoing PCI, ACEIs are known to have a better outcome than ARBs. However, there is limited evidence to suggest so. Among the STEMI registry consist of 1142 Korean patients, we compared the MACE, the composite of myocardial infarction, stoke, death, admission for heart failure, and target vessel revascularization, between the ACEI and ARB groups (Set 1). Further, we defined adequate medication as the administration of a dose equal to or higher than the initiation dose of ACEI according to the heart failure guideline recommendation with a mandatory addition of beta-blockers, and compared the outcomes between the inadequate and adequate medication groups (Set 2). Propensity score matching was used to eliminate difference. In the Set 1 comparison, patients in the ACEI group had a better outcome than those in the ARB group for both whole and matched populations (whole and matched population: Cox regression hazard ratio [HR], 0.645 and 0.535; 95% confidence interval [CI], 0.440-0.944 and 0.296-0.967; p = 0.024 and p = 0.039, respectively). In the Set 2 comparison for the whole population, patients in the inadequate medication group had more MACE than those in the adequate medication group (HR, 0.673; 95% CI, 0.459-0.985; p = 0.042). However, no difference was observed after propensity score matching (HR, 1.023; 95% CI, 0.654-1.602; p = 0.919). ACEIs might be a better choice than ARBs after primary revascularization. However, this study's findings suggest that early ACEI dose escalation combined with beta-blocker use may not improve prognosis.


A new method for the removal and analysis of small particles adhering to carpet fiber surfaces.
Fibers are a commonly encountered evidence type and fiber comparisons are routinely conducted. When the characteristics of questioned and known fibers correspond, the probative value is limited because the characteristics compared are determined by their manufacture. It would be valuable to test the hypothesis that questioned fibers originated from a specific carpet. One means of testing this is to analyze fine particles on the fibers. A procedure was developed for removing adhering particles from fibers and preparing them for scanning electron microscopy/ energy dispersive X-ray spectrometry (SEM/EDS) analysis. The efficacy of several fluids (acetone, ethanol, hexanes, aqueous ethanol, aqueous sodium hexametaphosphate) and agitation methods (sonication and vortexing) for removing particles from fibers was evaluated. The most effective method evaluated was sonication in ethanol for 10 min. The recovered particles were filtered onto a polycarbonate filter and analyzed by automated SEM/EDS analysis to generate data on the particle assemblage. This method makes possible future research to test the within-item and between-item variability of particle assemblages on fibers.


m3 Muscarinic receptor-induced and Gi-mediated heterologous potentiation of phospholipase C stimulation: role of phosphoinositide synthesis.
Agonist activation of thrombin and purinergic receptors endogenously expressed in human embryonic kidney (HEK) cells and of the stably expressed m3 muscarinic acetylcholine receptor (mAChR) induces phospholipase C (PLC) stimulation, with the most pronounced PLC stimulation observed on mAChR activation. These receptor responses were pertussis toxin (PTX) insensitive and nonadditive, suggesting that the receptors share common signaling pathways. Short term (2 min) pretreatment of HEK cells with carbachol (1 mM), but not ATP, followed by agonist washout, caused a long-lasting (> or = 90 min) sensitization of PLC responses. At 30 min after carbachol treatment and washout, mAChR-stimulated PLC activity, measured as formation of either total inositol phosphates or of inositol-1,4,5-trisphosphate, was enhanced by 1.5-2-fold. PLC stimulation by thrombin and purinergic receptors was increased by approximately 3-fold. Furthermore, carbachol pretreatment also enhanced, by approximately 2.5-fold, stimulation of PLC activity on direct activation of G proteins by AIF4- and guanosine-5'-O-(3-thio)-triphosphate in intact and permeabilized cells, respectively. In contrast, PLC activities, measured with exogenous phosphatidylinositol-4,5-bisphosphate [Ptdns(4,5)P2] in HEK cell lysates, were not altered, suggesting that carbachol pretreatment may enhance the cellular level of Ptdlns(4,5)P2. Indeed, the level of Ptdlns(4,5)P2 was found to be increased by approximately 50% in HEK cells 30 min after short term carbachol treatment, whereas the level of phosphatidylinositol was not altered and that of phosphatidylinositol-4-phosphate decreased (by 40-50%). Pretreatment of HEK cells with PTX prevented the m3 mAChR-induced PLC potentiation and reduced the elevation in Ptdlns(4,5)P2 level by approximately 50%. In conclusion, short term agonist activation of m3 mAChRs stably expressed in HEK cells can lead to a longlasting heterologous potentiation of PLC signaling, which processes apparently involve PTX-sensitive G proteins and an enhanced PLC substrate supply.


Genomic risk scores and oral contraceptive-associated ischemic stroke risk: a call for collaboration.
Oral contraceptives (OCs) are generally safe but vascular risk factors increase OC-associated ischemic stroke risk. We performed a case-control study to evaluate whether a genomic risk score for ischemic stroke modifies OC-associated ischemic stroke risk. The Genetics of Early-Onset Stroke study includes 332 premenopausal women (136 arterial ischemic stroke cases and 196 controls) with data on estrogen-containing OC use within 30 days before the index event (for cases) or interview (for controls). Using a previously validated genetic risk score (metaGRS) for ischemic stroke based on 19 polygenic risk scores for stroke and stroke-associated risk factors, we stratified our combined case-control sample into tertiles of genomic risk. We evaluated the association between OC use and ischemic stroke within each tertile. We tested if the association between OC use and ischemic stroke depended on the genomic risk of stroke using logistic regression with an OC use × metaGRS interaction term. These analyses were performed with and without adjustment for smoking, hypertension, diabetes, coronary heart disease, and body mass index. After adjustment for vascular risk factors, the odds ratio of OC use was 3.2 (1.7-6.3) overall and increased from the lower, middle, and upper tertile of genomic risk from 1.6 (0.5-5.4) to 2.5 (0.08-8.2) to 13.7 (3.8-67.3) respectively, and a p-value for interaction of 0.001. Our results suggest that genomic profile may modify the OC-associated ischemic stroke risk. Larger studies are warranted to determine whether a genomic risk score could be clinically useful in reducing OC-associated ischemic stroke.


Design and implementation of an automated partial volume correction in PET: application to dopamine receptor quantification in the normal human striatum.
The considerable effort and potential lack of reproducibility of human-driven PET quantification and partial volume correction (PVC) can be alleviated by use of atlas-based automatic analysis. The present study examined the application of a new algorithm designed to automatically define 3-dimensional regions of interest (ROIs) and their effect on dopamine receptor quantification in the normal human brain striatum, both without and with PVC. A total of 90 healthy volunteers (age range, 18-46 y) received a single injection of (11)C-raclopride, and automatic segmentation of concomitant structural MR images was performed using a maximum-probability atlas in combination with a trained neural network. For each identified tissue segment considered homogeneous for the tracer (or volumes of interest [VOIs]), an a priori criterion based on minimum axial recovery coefficient (RC(zmin) = 50%, 75%, and 90%) was used to constrain the extent of each ROI. With ROIs essentially overlapping the entire VOI volume (obtained with RC(zmin) = 50%), the binding potential (BP(ND)) of (11)C-raclopride was found to be around 2.2 for caudate and 2.9 for putamen, an underestimation by 35% and 28%, respectively, according to PVC values. At increased RC(zmin), BP(ND) estimates of (11)C-raclopride were increased by 12% and 21% for caudate and 8% and 15% for putamen when the associated ROIs decreased to around 65% and 43% of total tissue volume (VOI) for caudate and 67% and 31% for putamen. After PVC, we observed relative increases in BP(ND) variance of 12% for caudate and 20% for putamen, whereas estimated BP(ND) values all increased to 3.4 for caudate and 4.0 for putamen, regardless of ROI size. Dopamine receptor concentrations appeared less heterogeneous in the normal human striatum after PVC than they did without PVC: the 25%-30% difference in BP(ND) estimates observed between caudate and putamen remained significant after PVC but was reduced to slightly less than 20%. Furthermore, the results were comparable with those obtained with a manual method currently in use in our laboratory. The new algorithm allows for traditional PET data extraction and PVC in an entirely automatic fashion, thus avoiding labor-intensive analyses and potential intra- or interobserver variability. This study also offers the first, to our knowledge, large-scale application of PVC to dopamine D(2)/D(3) receptor imaging with (11)C-raclopride in humans.


Evaluation of zebrafish embryos as a model for assessing inhibition of hERG.
It has been proposed that the analysis of heart rate in zebrafish embryos can be used to assess the potential effects of compounds on hERG. The purpose of this study was to investigate the effect of compounds on the heart rate and atrioventricular dissociation in zebrafish. The compounds investigated were chosen based on the association or lack of association with QTc prolongation in the clinic. Three-day-old embryos were incubated in buffered embryo medium. On the day of the study, fish were placed in a petri dish containing 5.0 mL of embryo medium and 125 mg/L MS-222 anesthetic. Drugs to be tested were added to the medium from a stock solution to achieve the desired target concentration. Ten fish were incubated in each concentration of drug for 80 min. Beat rates of the atrium and ventricle were recorded after the incubation by counting beats of the respective chambers using standard brightfield stereomicroscopy. All of the compounds associated with QT prolongation induced dissociation between the atrium and ventricular rates except D,L-sotalol and procainamide. The concentrations that induced dissociation tended to be higher than the hERG IC50. None of the negative control compounds caused atrioventricular dissociation at clinically efficacious concentrations. In conclusion, the present data demonstrate that zebrafish can be utilized to assess the effects of chemicals on hERG. However, the practical use of this assay may be difficult because of the high concentrations that must be reached to see those pharmacological effects.


Developmental divergence of structural brain networks as an indicator of future cognitive impairments in childhood brain injury: Executive functions.
Brain insults during childhood can perturb the already non-linear trajectory of typical brain maturation. The diffuse effects of injury can be modelled using structural covariance networks (SCN), which change as a function of neurodevelopment. However, SCNs are estimated at the group-level, limiting applicability to predicting individual-subject outcomes. This study aimed to measure the divergence of the brain networks in paediatric traumatic brain injury (pTBI) patients and controls, and investigate relationships with executive functioning (EF) at 24 months post-injury. T1-weighted MRI acquired acutely in 78 child survivors of pTBI and 33 controls underwent 3D-tissue segmentation to estimate cortical thickness (CT) across 68 atlas-based regions-of-interest (ROIs). Using an 'add-one-patient' approach, we estimate a developmental divergence index (DDI). Our approach adopts a novel analytic framework in which age-appropriate reference networks to calculate the DDI were generated from control participants from the ABIDE dataset using a sliding-window approach. Divergence from the age-appropriate SCN was related to reduced EF performance and an increase in behaviours related to executive dysfunctions. The DDI measure showed predictive value with regard to executive functions, highlighting that early imaging can assist in prognosis for cognition.


Isolation and characterization of adenovirus messenger ribonucleic acid in productive infection.
Messenger ribonucleic acid (mRNA) from cells productively infected with adenovirus type 2 was isolated by affinity chromatography on polyuridylic acid [poly (U)] bound to Sepharose. At least 90% of the polyadenylic acid [poly (A)]-containing polysomal mRNA was retained by the poly (U) Sepharose and thus separated from more than 95% of the ribosomal RNA and transfer RNA. In these experiments, 65% of the early (3 to 5 hr postinfection) and 85% of the late (14 to 16 hr postinfection) virus-specific RNA was retained by the poly (U) Sepharose. Early in the infection 18%, and late in the infection more than 95%, of the poly (A)-containing fraction, eluted from the poly (U) Sepharose with 90% formamide, was adenovirus-specific, as shown by exhaustive hybridization. Different patterns, containing several distinct species of viral mRNA, were detected early and late in the infectious cycle. No distinct viral mRNA lacking poly (A) was discovered.


Mitochondrial abnormalities in Alzheimer's disease.
The finding that oxidative damage, including that to nucleic acids, in Alzheimer's disease is primarily limited to the cytoplasm of susceptible neuronal populations suggests that mitochondrial abnormalities might be part of the spectrum of chronic oxidative stress of Alzheimer's disease. In this study, we used in situ hybridization to mitochondrial DNA (mtDNA), immunocytochemistry of cytochrome oxidase, and morphometry of electron micrographs of biopsy specimens to determine whether there are mitochondrial abnormalities in Alzheimer's disease and their relationship to oxidative damage marked by 8-hydroxyguanosine and nitrotyrosine. We found that the same neurons showing increased oxidative damage in Alzheimer's disease have a striking and significant increase in mtDNA and cytochrome oxidase. Surprisingly, much of the mtDNA and cytochrome oxidase is found in the neuronal cytoplasm and in the case of mtDNA, the vacuoles associated with lipofuscin. Morphometric analysis showed that mitochondria are significantly reduced in Alzheimer's disease. The relationship shown here between the site and extent of mitochondrial abnormalities and oxidative damage suggests an intimate and early association between these features in Alzheimer's disease.


Uptake, translocation and metabolism of decabromodiphenyl ether (BDE-209) in seven aquatic plants.
Terrestrial plant uptake of PBDEs from contaminated soils has been widely reported recently. In this study the fate of deca-BDE within a plant/PBDEs/aquatic environment system was investigated through simulated pot experiments. Accumulations of the total PBDEs and deca-BDE were observed in tissues of seven test aquatic plant species, namely Phragmites australis, Cyperus papyrus, Alternanthera philoxeroides, Colocasia esculenta, Scirpus validus, Acorus calamus and Oryza sativa. In all seven plants, O. sativa leads the uptake and accumulation both in the total PBDEs (444.8 ng g(-1)) and deca-BDE (368.0 ng g(-1)) in roots. Among the six common phytoremediation aquatic plants, A. calamus leads the uptake (236.2 ng g(-1)), and P. australis leads the translocation (Cshoot/Croot = 0.35), while A. philoxeroides (43.4%) and P. australis (80.0%) lead in the metabolism efficiencies in the root and shoot, respectively. The detection of seventeen lesser brominated PBDE congeners provided the debromination evidence, and the specific PBDEs profiles in test plant species indicated there is no common metabolic pattern. Furthermore, a relative high proportion of lesser brominated PBDE congeners in shoots suggested the possible metabolic difference between roots and shoots. Finally, a noticeable percentage of penta- and octa-BDE derived from deca-BDE also hint the ecological risk in deca-BDE use. This comparative research on the aquatic plants provide a broad vision on the understanding of plant/PBDEs/aquatic environment interaction system, and may be applied to remediate PBDEs in contaminated waters and sediments.


Adaptive classifier integration for robust pattern recognition.
The integration of multiple classifiers promises higher classification accuracy and robustness than can be obtained with a single classifier. This paper proposes a new adaptive technique for classifier integration based on a linear combination model. The proposed technique is shown to exhibit robustness to a mismatch between test and training conditions. It often outperforms the most accurate of the fused information sources. A comparison between adaptive linear combination and non-adaptive Bayesian fusion shows that, under mismatched test and training conditions, the former is superior to the latter in terms of identification accuracy and insensitivity to information source distortion.


A Polymorphism within the Promoter of the Dopamine Receptor D1 (DRD1 -48A/G) Associates with Impaired Kidney Function in White Hypertensive Patients.
Dopamine DRD1 receptor regulates renal function and vascular resistance. It plays a role in the pathogenesis of hypertension in animal models. In humans, the DRD1 gene presents a A-48G polymorphism associated to hypertension in a Japanese population. To explore the role of this polymorphism on blood pressure and renal function in Caucasian hypertensive patients (H), we evaluated the allele frequencies in a populations of 697 H and 100 blood volunteers, and found no difference in the distribution of the alleles between the two groups (AA;AG;GG: 13%;50%;37%; and 12%; 51%;36% respectively). In H, we found a significant difference between AA and GG in serum creatinine (AA: 1.06±.08 mg/dl; GG:0.97±0.02 mg/dl, p<0.03). Treatment restored serum creatinine at levels comparable between genotypes (AA: 0.99±0.03 mg/dl; GG: 0.94±0.02 mg/dl, n.s.). To replicate the finding, in a case control study of 8 AA and 7 GG hypertensive patients matched for age, sex and body mass index, in pharmacological wash out for 30 days, we evaluated serum (Creatinine, Na, Uric Acid, Urea) and urinary (volume/24h, protein/24h, creatinine clearance/24h) biochemistry and renal hemodynamic assessed by ultrasound. Once again, the AA group showed higher serum creatinine, Na, Uric acid and urea, reduced creatinine clearance and a higher level of urinary protein excretion. These changes occurred while no differences were observed in diuresis and renal vascular resistances. In conclusions, the DRD1 A-48G polymorphism identifies a class of H that is prone to hypertension related kidney alterations.


Mortality in trauma patients: the interaction between host factors and severity.
Data on host factors influencing mortality in trauma patients is sparse and contradictory. To develop a model for health policy decisions, we examined all trauma admissions to acute care hospitals in the state of California in the year 1986. We looked at the influence of the following host factors: age, gender, and preinjury medical conditions, on mortality stratified by injury severity. The study group (N = 199,737) had an overall mortality rate of 1.9%. Mortality increased starting at age 40 years and was independently influenced by gender, the presence of pre-existing disease, and the body region injured. In patients with minor injury, mortality rates became higher in the elderly at age 65+. However, in patients with injuries of moderate severity, mortality increased in both middle age (40-64) and elderly groups (65+). Male gender was also a risk factor, present in both the elderly and middle age groups. While the presence of both pre-existing medical disease or injury to head or abdomen was related to increased mortality in middle-aged patients at all severity levels, neither accounted for the effect of gender. Conclusion. Age and gender influence mortality in trauma patients. These effects are not explained by documented pre-existing disease or region of injury. Age and gender serve only as observable markers for subgroups of patients with impaired response to injury. Middle-aged males comprise a previously unrecognized high-risk subgroup for this impaired response.


Predictors of outcome in acute traumatic central cord syndrome due to spinal stenosis.
the objective of this study was to elucidate the relationship between admission demographic data, validated injury severity measures on imaging studies, and clinical indicators on the American Spinal Injury Association (ASIA) motor score, Functional Independence Measure (FIM), manual dexterity, and dysesthetic pain at least 12 months after surgery for acute traumatic central cord syndrome (ATCCS) due to spinal stenosis. over a 100-month period (January 2000 to April 2008), of 211 patients treated for ATCCS, 59 cases were due to spinal stenosis, and these patients underwent surgical decompression. Five of these patients died, 2 were lost to follow-up, 10 were not eligible for the study, and the remaining 42 were followed for at least 12 months. in the cohort of 42 patients, mean age was 58.3 years, 83% of the patients were men, and 52.4% of the accidents were due to falls. Mean admission ASIA motor score was 63.8 (upper extremities score, 25.8 and lower extremities score, 39.8), the spinal cord was most frequently compressed at skeletal segments C3-4 and C4-5 (71%), mean midsagittal diameter at the point of maximum compression was 5.6 mm, maximum canal compromise (MCC) was 50.5%, maximum spinal cord compression was 16.5%, and length of parenchymal damage on T2-weighted MR imaging was 29.4 mm. Time after injury until surgery was within 24 hours in 9 patients, 24-48 hours in 10 patients, and more than 48 hours in 23 patients. At the 1-year follow-up, the mean ASIA motor score was 94.1 (upper extremities score, 45.7 and lower extremities score, 47.6), FIM was 111.1, manual dexterity was 64.4% of baseline, and pain level was 3.5. Stepwise regression analysis of 10 independent variables indicated significant relationships between ASIA motor score at follow-up and admission ASIA motor score (p = 0.003), MCC (p = 0.02), and midsagittal diameter (p = 0.02); FIM and admission ASIA motor score (p = 0.03), MCC (p = 0.02), and age (p = 0.02); manual dexterity and admission ASIA motor score (p = 0.0002) and length of parenchymal damage on T2-weighted MR imaging (p = 0.002); and pain level and age (p = 0.02) and length of parenchymal lesion on T2-weighted MR imaging (p = 0.04). the main indicators of long-term ASIA motor score, FIM, manual dexterity, and dysesthetic pain were admission ASIA motor score, midsagittal diameter, MCC, length of parenchymal damage on T2-weighted MR imaging, and age, but different domains of outcome were determined by different predictors.


Exploring the norepinephrine to angiotensin II conversion ratio in patients with vasodilatory hypotension: A post-hoc analysis of the ARAMIS trial.
Angiotensin II is approved for catecholamine-refractory vasodilatory shock but the conversion dose ratio from norepinephrine to angiotensin II remains unclear. We conducted a post-hoc analysis of the Acute Renal effects of Angiotensin II Management in Shock (ARAMIS) trial involving patients with vasodilatory hypotension. We determined the norepinephrine equivalent dose immediately prior to angiotensin II initiation and calculated the conversion dose ratio between norepinephrine and angiotensin II. We performed subgroup analyses based on recent exposure to angiotensin receptor blockers (ARBs) and renin levels at baseline. In 37 patients, the median conversion dose ratio between norepinephrine equivalent and angiotensin II was to 10:1 for norepinephrine bitartrate (5:1 for norepinephrine base). The conversion ratio was not affected by the baseline renin, with a median ratio of 10 (7-21) in the high renin group versus 12 (5-22) in the low renin group. Finally, exposure to ARBs prior admission appeared to diminish the conversion ratio with a median ratio of 7 (4-13) in ARB patients vs. 12 (7-22) in non-ARB patients. The norepinephrine to angiotensin II conversion dose ratio is 10:1 in a vasodilatory hypotension population. These findings can guide clinicians and researchers in the use, dosing, and study of angiotensin II in critical care.


Surgical management of bronchopulmonary malformations.
Bronchopulmonary malformations (BPM) are rare conditions, which typically arise below the carina and can result in significant morbidity (infection and/or hemorrhage) and mortality (respiratory failure). All children with BPM surgically treated from 2001-2014 at a tertiary care children's hospital were identified. Patient demographics, surgical indications, procedure type, estimated blood loss, pathology, perioperative complications, length of stay, and outcomes were analyzed. A total of 41 BPM patients underwent surgery with 98% overall survival (one abdominal BPM expired) but 100% for thoracic lesions. Resections were performed thoracoscopically (37%), thoracoscopy converted to open (22%), and via thoracotomy (37%). Poor visualization (67%) or inability to tolerate single lung ventilation (33%) led to conversions. No conversions resulted from hemorrhage or received blood transfusions. Patients with prenatally diagnosed BPM were more likely to undergo thoracoscopic surgery (odds ratio [OR], 18.2) versus nonprenatally diagnosed, P = 0.002. Open/converted patients had longer chest tube days (6.2) versus thoracoscopic (2.9), P = 0.048. Additionally, respiratory distress was a more common indication in patients aged <4 mo (OR, 28.0) versus ≥4 mo and <6 kg (OR, 40.5) versus ≥6 kg, P < 0.001. Open resections were more common in patients aged <4 mo (OR, 26.3) versus ≥4 mo, P = 0.002. Operative time was shorter and estimated blood loss (mL/kg) was greater for <6 versus ≥6 kg, P < 0.05. BPM resections have high overall survival. Chest tube days are shorter among thoracoscopic patients, but conversion to thoracotomy can avoid hemorrhage and need for transfusion. Size and respiratory distress limit use of thoracoscopy in young infants with BPM.


[Hereditary progressive mucinous histiocytosis].
Hereditary progressive mucinous histiocytosis is an uncommon condition which to date has only been observed in women. The disease is a non-Langerhans histiocytosis with a stereotypic clinical presentation limited to the skin. The clinical, histological and genetic features of this disease differ considerably from other types of histiocytoses and overload diseases. We report the first case observed in France. A 49-year-old woman consulted for asymptomatic papules on the dorsal aspect of the hands known since childhood. The lesions had progressively spread to other regions of the body. The woman's mother and two sisters had the same condition but her two sons and her brother and nephews did not. Standard semi-thin slices and electron microscope histology demonstrated dense cellular infiltration of the reticular dermis with interstitial mucinous overload, voluminous histiocytes with characteristic toluidine blue positive granulations, and a cytoplasm rich in myelin and Zebra bodies respectively. The observed clinical, histological and genetic features were characteristic of progressive hereditary progressive mucinous histiocytosis. The pedigree in our case and in those reported in the literature suggest a dominant hereditary condition. The fact that only female cases have been reported to date would suggest X-linked transmission with early death of male fetuses. An autosomal dominant transmission limited to woman as a result of mitochondrial heredity or hormone-related phenomena cannot be ruled out. The progressive extension with age without a tendency for spontaneous regression and the electron microscopy aspect suggest a phospholipid deposit disorder similar to that observed in lysosomial diseases. However, primary macrophage proliferation in response to an unknown stimulus cannot be excluded.


Simultaneous occurrence of folliculitis decalvans capillitii in identical twins.
Folliculitis decalvans is a chronic purulent folliculitis resulting in permanent hair loss and follicular atrophy. We report 32-year-old identical female twins presenting with relapsing pruritic outbreaks on the scalp resulting in areas of permanent baldness. Staphylococcus aureus was detected in the lesions of both women. Histopathology confirmed the diagnosis of folliculitis decalvans. Immunological testing showed no alteration of the immune system. To our knowledge, this is the first report on folliculitis decalvans occurring in identical twins, suggesting a possible genetic component in this disease.


Visualizing seismic risk and uncertainty: a review of related research.
Government agencies and other authorities often communicate earthquake risks using maps derived from geographic information systems. Yet, little is known about the effects of these maps on risk perceptions. While mental models research and other approaches are available to inform risk communication text design, similar empirically derived guidance is lacking for visual risk communications, such as maps, which are likely to trump text in their impact and appeal. This paper reviews the empirical research that might inform such guidance. Research on graphs, spatial and visual perception, and map design suggests that graphics increase risk avoidance over numerical risk representations, and countable visuals, like dots, can increase the accuracy of perceived risks, but not always. Cartographic design features, such as color, animation, interactivity, and depth cues, are all candidates to represent risk and uncertainty and to influence risk perception. While there are robust known effects of color (e.g., red = danger), with some cultural variability, animation can increase the salience of otherwise obscure features but is not uniformly effective. Depth cues, dimensionality, and the extent to which a representation depicts versus symbolizes a scene will influence the viewer's perspective and perception, depending on the viewer's familiarity with the scene; their effects on risk perception remain unclear. The translation and representation of technical information about risk and uncertainty is critical to risk communication effectiveness. Our review suggests a handful of candidate criteria for evaluating the effects of risk visualizations, short of changes in behavior: accuracy, accessibility, retention, and perceived risk and usefulness.


Intensive glucose control in the management of diabetes mellitus and inpatient hyperglycemia.
The current evidence on intensive glycemic control in the inpatient and outpatient settings and its implications to practice are reviewed. Poor glycemic control in patients with diabetes is associated with microvascular and macrovascular complications. Various clinical trials involving patients with type 1 and type 2 diabetes have revealed the benefits of intensive glycemic control in delaying the onset and progression of microvascular complications of diabetes. However, while long-term epidemiologic trials and a meta-analysis have shown a benefit of intensive glycemic control in reducing the incidence of macrovascular complications, recent clinical trials have not found similar benefits. The American Diabetes Association (ADA), American College of Endocrinology (ACE), and American Association of Clinical Endocrinologists recommend intensive control of glycosylated hemoglobin and plasma glucose at specified goals. Hyperglycemia in the inpatient setting is associated with increased morbidity and mortality. ACE and ADA recommend the use of an i.v. insulin infusion in critically ill inpatients with hyperglycemia. In noncritically ill inpatients, basal and bolus doses of insulin are recommended. The use of sliding-scale insulin as the sole therapy for inpatient hyperglycemia is discouraged. However, caution must be exercised to ensure a balance between controlling hyperglycemia and reducing the risk of hypoglycemia. While intensive glycemic control is known to prevent or delay the occurrence of microvascular complications of diabetes, macrovascular benefits are still uncertain. Current evidence suggests that intensive glycemic control should be initiated as soon as possible after diagnosis of type 1 or type 2 diabetes in order to maximize potential long-term macrovascular benefits. Inpatient hyperglycemia should be managed appropriately to reduce morbidity and mortality, with great care taken to avoid and appropriately treat hypoglycemia.


Comparative effects of substituted phenylethylamines on brain serotonergic mechanisms.
A series of substituted phenylethylamines were examined for their relative potencies to 1) release and inhibit accumulation of labeled serotonin in brain synaptosomes, 2) compete with serotonin receptor binding and 3) induce the "serotonin syndrome" in mice. In general, the relative potencies of the phenylethylamine derivatives to produce the serotonin syndrome did not correlates well with their effects on synaptosomal uptake or release of serotonin or serotonin receptor binding. However, para chloro substitution was more effective than meta or ortho chlorophenylethylamine and para methoxy substitution more effective than ortho methoxyphenylethylamine for [3H]serotonin uptake inhibition and release and induction of the serotonin syndrome. Lineweaver Burk kinetic analysis indicated that para chlorophenylethylamine was a competitive inhibitor of [3H]serotonin uptake in vitro with a Ki of 4.3 x 10(-7) M. Fluoxetine, a specific serotonin uptake inhibitor, blocked p-chlorophenylethylamine and p-methoxyphenylethylamine-induced [3H]serotonin release. The ability of phenylethylamine derivatives to release preferentially [3H]serotonin in vitro without damaging serotonergic neurons and to consistently and rapidly produce the serotonin syndrome in vivo should make these compounds useful pharmacological tools for investigating brain serotonin metabolism.


Contribution of V(H) replacement products in mouse antibody repertoire.
VH replacement occurs through RAG-mediated recombination between the cryptic recombination signal sequence (cRSS) near the 3' end of a rearranged VH gene and the 23-bp RSS from an upstream unrearranged VH gene. Due to the location of the cRSS, VH replacement leaves a short stretch of nucleotides from the previously rearranged VH gene at the newly formed V-D junction, which can be used as a marker to identify VH replacement products. To determine the contribution of VH replacement products to mouse antibody repertoire, we developed a Java-based VH Replacement Footprint Analyzer (VHRFA) program and analyzed 17,179 mouse IgH gene sequences from the NCBI database to identify VH replacement products. The overall frequency of VH replacement products in these IgH genes is 5.29% based on the identification of pentameric VH replacement footprints at their V-D junctions. The identified VH replacement products are distributed similarly in IgH genes using most families of VH genes, although different families of VH genes are used differentially. The frequencies of VH replacement products are significantly elevated in IgH genes derived from several strains of autoimmune prone mice and in IgH genes encoding autoantibodies. Moreover, the identified VH replacement footprints in IgH genes from autoimmune prone mice or IgH genes encoding autoantibodies preferentially encode positively charged amino acids. These results revealed a significant contribution of VH replacement products to the diversification of antibody repertoire and potentially, to the generation of autoantibodies in mice.


[The effect of lithium oxybutyrate on aggressive animal behavior].
The aim of the study was to examine the aggressive behavior of rats after lithium hydroxybutyrate injections (10 mg/kg once a day, at 9 a.m or 7 p.m., for a week). It was found that the drug did not affect the aggressive behavior of isolated rats towards mice placed to them. However, in the evening the drug inhibited aggressive manifestations (attacks, bites) in rats with raphe midbrain lesions. The 5-hydroxytryptophan (20 mg/kg, intraperitoneally, once) augmented the antiaggressive properties of lithium hydroxybutyrate in the evening but weakened them in the morning.


The Effect of Age on the Relative Outcomes of Cemented and Cementless Mobile-Bearing Unicompartmental Knee Arthroplasty, Based on Data From National Databases.
Unicompartmental knee arthroplasty (UKA) is an effective treatment for medial compartment arthritis. A challenge is that patients requiring knee arthroplasty are becoming younger. It is currently unknown what the relative performances of cemented and cementless UKAs are, in different age groups. A total of 12,882 cemented and cementless UKAs from the National Joint Registry and Hospital Episodes Statistics databases were matched on patient and surgical factors. Patients were stratified into 3 groups: (1) <60 years; (2) 60-69 years; and (3) ≥70 years. Revision and reoperation rates were compared using Cox regression analyses. The 10-year implant survival for the matched cemented and cementless UKAs for (1) <60 years were 81.4% (CI 73.6-87.0) and 86.7% (CI 80.7-90.9) (hazard ratio [HR] 0.73, CI 0.56-0.94, P = .02); (2) for 60-69 years were 91.8% (CI 88.9-94.0) and 94.5% (CI 92.9-95.7) (HR 0.90, CI 0.67-1.22, P = .51); and (3) ≥70 years were 93.5% (CI 91.1-95.3) and 94.2% (CI 92.0-95.8) (HR 1.0, CI 0.71-1.40, P = .99). The same trend was observed for reoperations. In the <60 years and 60-69 years groups there were significantly fewer revisions for aseptic loosening in the cementless group (0.5% versus 1.6% [P < .001] and 0.4% versus 1.3% [P = .002], respectively). Younger ages were associated with higher revision rates in both cemented and cementless UKA groups. Cementless fixation has reduced long-term revision rates compared to cemented fixation in the <60 years group with aseptic loosening rates 3 times lower. III.


Synthetic peracetate tea polyphenols as potent proteasome inhibitors and apoptosis inducers in human cancer cells.
It has been suggested that proteasome activity is essential for tumor cell proliferation and drug resistance development. We have previously shown that natural and synthetic ester bond-containing tea polyphenols are selective inhibitors of the chymotrypsin-like activity of the proteasome. The most abundant catechin in green tea is (-)-epigallocatechin-3-gallate [(-)-EGCG], which has been found by many laboratories to exhibit the most potent anticancer activity. We have reported that (-)-EGCG is also the most effective proteasome inhibitor among all the natural green tea catechins tested. Unfortunately, (-)-EGCG is very unstable in neutral and alkaline conditions. In an attempt to increase the stability and thus the efficacy, we synthesized several (-)-EGCG analogs with acetyl protected -OH groups as prodrugs. Here we report, for the first time, that these acetylated synthetic tea analogs are much more potent than natural (-)-EGCG in inhibiting the proteasome in cultured tumor cells. Consistently, these protected analogs showed much higher potency than (-)-EGCG to inhibit proliferation and transforming activity and to induce apoptosis in human leukemic, prostate, breast, and simian virus 40-transformed cells. Additionally, these protected analogs had greatly reduced effects on human normal and non-transformed cells. Therefore, these peracetate protected tea polyphenols are more efficacious than (-)-EGCG and possess great potential to be developed into novel anticancer drugs. Identification of the cytosolic metabolite(s) of peracetate-protected polyphenols in cultured tumor cells and examination of their in vivo tumor growth-inhibitory activity are currently underway in our laboratory.


Repetitive intratracheal bleomycin models several features of idiopathic pulmonary fibrosis.
Single-dose intratracheal bleomycin has been instrumental for understanding fibrotic lung remodeling, but fails to recapitulate several features of idiopathic pulmonary fibrosis (IPF). Since IPF is thought to result from recurrent alveolar injury, we aimed to develop a repetitive bleomycin model that results in lung fibrosis with key characteristics of human disease, including alveolar epithelial cell (AEC) hyperplasia. Wild-type and cell fate reporter mice expressing β-galactosidase in cells of lung epithelial lineage were given intratracheal bleomycin after intubation, and lungs were harvested 2 wk after a single or eighth biweekly dose. Lungs were evaluated for fibrosis and collagen content. Bronchoalveolar lavage (BAL) was performed for cell counts. TUNEL staining and immunohistochemistry were performed for pro-surfactant protein C (pro-SP-C), Clara cell 10 (CC-10), β-galactosidase, S100A4, and α-smooth muscle actin. Lungs from repetitive bleomycin mice had marked fibrosis with prominent AEC hyperplasia, similar to usual interstitial pneumonia (UIP). Compared with single dosing, repetitive bleomycin mice had greater fibrosis by scoring, morphometry, and collagen content; increased TUNEL+ AECs; and reduced inflammatory cells in BAL. Sixty-four percent of pro-SP-C+ cells in areas of fibrosis expressed CC-10 in the repetitive model, suggesting expansion of a bronchoalveolar stem cell-like population. In reporter mice, 50% of S100A4+ lung fibroblasts were derived from epithelial mesenchymal transition compared with 33% in the single-dose model. With repetitive bleomycin, fibrotic remodeling persisted 10 wk after the eighth dose. Repetitive intratracheal bleomycin results in marked lung fibrosis with prominent AEC hyperplasia, features reminiscent of UIP.


[Prevalence and intensity of dental diseases among foreign students during their study in Russia].
The findings of complex dental study of 235 1-st and 2-nd year foreign students of TSMA were given in the article. High level of prevalence and intensity of dental diseases was found. The obtained results of the study should be used to develop measures for their future treatment and to prevent dental diseases.