Dichloroacetate stabilizes the intraoperative acid-base balance during liver transplantation.
Lactic acidosis occurs during orthotopic liver transplantation (OLT), especially during the anhepatic and early postreperfusion phases. Dichloroacetate (DCA) inhibits pyruvate dehydrogenase kinase-1, indirectly activating mitochondrial pyruvate dehydrogenase. This, in turn, markedly reduces systemic lactate production and, to a lesser extent, increases hepatic lactate uptake. The result is moderation of lactic acidosis in many clinical conditions. This study evaluated the efficacy of DCA in controlling lactic acidosis during OLT and improving perioperative outcome from OLT. After informed consent, 250 patients for OLT received either intraoperative DCA or placebo. DCA (40 mg/kg intravenously) or placebo was administered after anesthesia induction and repeated 4 hours later. Intraoperative measures were arterial blood gases, lactate, and Na+ and utilization of blood products, CaCl2, and NaHCO3. Outcome measures were time to tracheal extubation, intensive care unit length of stay, hospital length of stay, requirement for postoperative plasma transfusion, retransplantation, and perioperative mortality. DCA reduced the arterial lactic acid concentration by an average of 44% (1.8 mmol L(-1), P < 0.001), stabilized the acid-base balance, and reduced NaHCO(3) administration by 80% (P < 0.001). Postoperatively, DCA-treated patients required 50% less postoperative plasma transfusion (2 versus 4 units, respectively, P = 0.016), but the incidence of transfusion was similar in both groups (62% versus 60%, P = 0.381). DCA did not alter time to extubation, intensive care unit length of stay, or hospital length of stay. In conclusion, DCA attenuated lactic acidosis during OLT, stabilizing the intraoperative acid-base balance and decreasing NaHCO3 use. DCA decreased postoperative plasma transfusion requirement but otherwise had no measurable effect on perioperative outcome parameters.


The RHOX homeobox gene cluster is selectively expressed in human oocytes and male germ cells.
What human tissues and cell types express the X-linked reproductive homeobox (RHOX) gene cluster? The RHOX homeobox genes and proteins are selectively expressed in germ cells in both the ovary and testis. The RHOX homeobox transcription factors are encoded by an X-linked gene cluster whose members are selectively expressed in the male and female reproductive tract of mice and rats. The Rhox genes have undergone strong selection pressure to rapidly evolve, making it uncertain whether they maintain their reproductive tissue-centric expression pattern in humans, an issue we address in this report. We examined the expression of all members of the human RHOX gene cluster in 11 fetal and 8 adult tissues. The focus of our analysis was on fetal testes, where we evaluated 16 different samples from 8 to 20 weeks gestation. We also analyzed fixed sections from fetal testes, adult testes and adult ovaries to determine the cell type-specific expression pattern of the proteins encoded by RHOX genes. We used quantitative reverse transcription-polymerase chain reaction analysis to assay human RHOX gene expression. We generated antisera against RHOX proteins and used them for western blotting, immunohistochemical and immunofluorescence analyses of RHOXF1 and RHOXF2/2B protein expression. We found that the RHOXF1 and RHOXF2/2B genes are highly expressed in the testis and exhibit low or undetectable expression in most other organs. Using RHOXF1- and RHOXF2/2B-specific antiserum, we found that both RHOXF1 and RHOXF2/2B are primarily expressed in germ cells in the adult testis. Early stage germ cells (spermatogonia and early spermatocytes) express RHOXF2/2B, while later stage germ cells (pachytene spermatocytes and round spermatids) express RHOXF1. Both RHOXF1 and RHOXF2/2B are expressed in prespermatogonia in human fetal testes. Consistent with this, RHOXF1 and RHOXF2/2B mRNA expression increases in the second trimester during fetal testes development when gonocytes differentiate into prespermatogonia. In the human adult ovary, we found that RHOXF1 and RHOXF2/2B are primarily expressed in oocytes. While the average level of expression of RHOX genes was low or undetectable in all 19 human tissues other than testes, it is still possible that RHOX genes are highly expressed in a small subset of cells in some of these non-testicular tissues. As a case in point, we found that RHOX proteins are highly expressed in oocytes within the human ovary, despite low levels of RHOX mRNA in the whole ovary. The cell type-specific and developmentally regulated expression pattern of the RHOX transcription factors suggests that they perform regulatory functions during human fetal germ cell development, spermatogenesis and oogenesis. Our results also raise the possibility that modulation of RHOX gene levels could correct some cases of human infertility and that their encoded proteins are candidate targets for contraceptive drug design.


Histological investigation of the effect of soybean (Glycine max) extracts on the collagen layer and estrogen receptors in the skin of female rats.
The purpose of this study was to analyze the effects of soybean extracts obtained using different extraction methods on the skin of female rats. A total of 64 female Sprague-Dawley rats were divided into 8 equal groups. Various extracts were administered to the female rats by oral gavage for one month. The groups comprised carboxymethyl cellulose-free control, carboxymethyl cellulose-plus control, 100-mg/kg n-hexane extract, 200-mg/kg n-hexane extract, 100-mg/kg ethyl acetate extract, 200-mg/kg ethyl acetate extract, 100-mg/kg ethanol extract and 200-mg/kg ethanol extract groups. The thickness of the collagen layer and the number of estrogen receptor-positive cells were evaluated. All the extract-treated groups showed a statistically significant decrease in the number of estrogen receptor-positive cells compared with the control groups. Regarding the thickness of the collagen layer, only the 200-mg/kg ethyl acetate extract-treated group showed a significant increase compared with the control groups (p<0.05). Our data suggest that oral intake of three different total soybean extracts might have positive estrogenic effects on the skin and that only a high-dose ethyl acetate extract can increase the expression of collagen, which may prove to be beneficial for postmenopausal facial skin.


Validation and Cultural Adaptation of Explanatory Model Interview Catalogue (EMIC) in Assessing Stigma among Recovered Patients with COVID-19 in Saudi Arabia.
Stigma is a negative feeling affecting many patients with various health conditions, especially the contagious ones such as COVID-19. The Explanatory Model Interview Catalogue (EMIC) is one of the valid and reliable stigma-measuring tools; however, it has not been translated and validated in Arabic. Therefore, the aim of this study was to translate and validate the EMIC in Arabic among a sample of Arabic-speaking adults who recently recovered from COVID-19 in Saudi Arabia. The 12 items of the EMIC scale were forward- and backward-translated and reviewed by all authors to check the face and content validity prior to approving the final version of the Arabic 12-item EMIC. A total of 174 participants aged ≥18 years who contracted COVID-19 and recovered as of 29 July 2020 were interviewed. The Cronbach's alpha of the Arabic version of the 12-item EMIC was 0.79, indicating an acceptable level of internal consistency. Using principal component analysis with varimax rotation, two factors explained more than 60% of the variance of the translated EMIC scale. The mean EMIC score was 5.91, implying a low level of stigma among participants. Married participants (β = 2.93; 95%CI 0.88 to 4.98, p = 0.005) and those with a family history of mental illness (β = 2.38; 95%CI 0.29 to 4.46, p = 0.025) were more likely to have higher EMIC scores in comparison to their counterparts who were unmarried and had no family history of mental illness. On the contrary, older adults were less likely to have high EMIC scores (β = -0.11; 95%CI -0.21 to -0.01, p = 0.03). Future studies with larger samples of patients with COVID-19 and various health conditions should be conducted to examine the validity and reliability of the Arabic version of the EMIC among different patient populations and to unveil the factors that may play a role in patients' feelings of stigmatization in this part of the world.


A ribonucleoprotein-based decaplex CRISPR/Cas9 knockout strategy for CHO host engineering.
Chinese hamster ovary (CHO) cell engineering based on CRISPR/Cas9 knockout (KO) technology requires the delivery of guide RNA (gRNA) and Cas9 enzyme for efficient gene targeting. With an ever-increasing list of promising gene targets, developing, and optimizing a multiplex gene KO protocol is crucial for rapid CHO cell engineering. Here, we describe a method that can support efficient targeting and KO of up to 10 genes through sequential transfections. This method utilizes Cas9 protein to first screen multiple synthetic gRNAs per gene, followed by Sanger sequencing indel analysis, to identify effective gRNA sequences. Using sequential transfections of these potent gRNAs led to the isolation of single cell clones with the targeted deletion of all 10 genes (as confirmed by Sanger sequencing at the DNA level and mass spectrometry at the protein level). Screening 704 single cell clones yielded 6 clones in which all 10 genes were deleted through sequential transfections, demonstrating the success of this decaplex gene editing strategy. This pragmatic approach substantially reduces the time and effort required to generate multiple gene knockouts in CHO cells.


Epstein-Barr virus-specific transcription in normal and malignant nasopharyngeal biopsies and in lymphocytes from healthy donors and infectious mononucleosis patients.
Cytoplasmic RNA was prepared from biopsy material obtained from the nasopharynx of normal individuals and nasopharyngeal carcinoma (NPC) patients. Similar RNA preparations were prepared from lymphocytes of healthy donors and infectious mononucleosis patients. RNA was radioactively labelled in vitro and hybridized to cloned fragments of B95-8 Epstein-Barr virus (EBV). In all seropositive cases the minimum pattern of EBV-specific RNA expression was like that observed previously in latently infected, EBV-positive lymphoblastoid cell lines or Burkitt's lymphoma biopsies. A novel observation was that all of the control biopsies from normal nasopharynxes expressed EBV-specific RNA, some of which appeared to be associated with a more active state of EBV infection. The pattern of expression in NPC patients was similar to that of the normal donors but showed some variations in complexity. In general the virus-specified small RNAs were not present in normal nasopharyngeal tissue but were expressed in the NPC biopsies.


Adverse drug reaction-related hospitalisations: a population-based cohort study.
To evaluate the extent, characteristics and determinants of adverse drug reaction (ADR)-related hospitalisations on a population-based level in 2003. We performed a cohort study in the Integrated Primary Care Information (IPCI) database, a general practitioners (GPs) research database with longitudinal data from electronic patient records of a group of 150 GP throughout the Netherlands. Hospital discharge letters and patient records were reviewed to evaluate ADR-related hospitalisations applying WHO causality criteria. The prevalence of ADR-related hospitalisations per total admissions and the incidence per drug group were calculated. Avoidability and seriousness of the ADRs causing admission were assessed applying the algorithm from Hallas. We identified 3515 hospital admissions, 1277 elective and 2238 acute. Of the acute admissions, 115 were caused by an ADR giving a prevalence of 5.1% (95% confidence intervals (CI): 4.3-6.1%). The prevalence of ADR-related acute admissions increased with age up to 9.8% (95%CI: 7.5-12.7) for persons >75 years. The ADRs that most frequently caused hospitalisations were gastro-intestinal bleeding with anti-thrombotics, bradycardia/hypotension with cardiovascular drugs and neutropenic fever with cytostatics. The incidence rate of ADR-related hospitalisations per drug group was highest for anti-thrombotics and anti-infectives and was relatively low for cardiovascular drugs. Fatality as a direct consequence of the ADR-related admission was 0.31%. In elderly patients 40% of the ADRs causing hospitalisation were judged to be avoidable. The extent and potential avoidability of ADR-related hospitalisations is still substantial, especially in elderly patients. Measures need to be put into place to reduce the burden of ADRs.


Mechanochromic and thermochromic shape memory photonic crystal films based on core/shell nanoparticles for smart monitoring.
Shape memory photonic crystals (SMPCs) combining the main characteristics of shape memory materials and photonic crystals have drawn increasing research interest. In sharp contrast to traditional responsive photonic crystals, the temporary shape of SMPCs can be "frozen" and photonic configurations can be modulated by temperature. However, the large-scale fabrication of SMPCs still remains a big challenge, making the practical application difficult. Herein novel scalable SMPC films with both mechanochromic and thermochromic properties are reported. Unlike traditional template-based methods resulting in only a small size, SMPC films are fabricated by a facile hot-pressing method and post-photocuring technology to give large-area freestanding polymer films. The films are mechanically robust and flexible, featuring an excellent structural color which can be changed upon stretching, similar to the color change process of chameleons in response to the environment. The blue-shift of the reflection peak up to 120 nm can be observed when the film is stretched. The films can be reversibly stretched and recovered in 25 cycles without obvious changes in reflection spectra. The temporary shape accompanied by tremendous color changes in the corresponding SMPC films after mechanical stress induced hot programming could be simply fixed by cooling the structure below the glass transition temperature of the polymer matrix. Incorporated programmed optical properties could afterwards be erased by temperature, and initial optical properties could be fully restored. Based on the fully reversible programmable shape as well as optical properties, the investigated SMPC films are expected to be promising candidates for various potential applications, such as smart monitoring, sensors, anti-counterfeiting, and displays.


Polyaniline nano-needles into electrospun bio active fibres support in vitro astrocyte response.
Recent studies have proposed that the bioelectrical response of glial cells, called astrocytes, currently represents a key target for neuroregenerative purposes. Here, we propose the fabrication of electrospun nanofibres containing gelatin and polyaniline (PANi) synthesized in the form of nano-needles (PnNs) as electrically conductive scaffolds to support the growth and functionalities of primary astrocytes. We report a fine control of the morphological features in terms of fibre size and spatial distribution and fibre patterning, i.e. random or aligned fibre organization, as revealed by SEM- and TEM-supported image analysis. We demonstrate that the peculiar morphological properties of fibres - i.e., the fibre size scale and alignment - drive the adhesion, proliferation, and functional properties of primary cortical astrocytes. In addition, the gradual transmission of biochemical and biophysical signals due to the presence of PnNs combined with the presence of gelatin results in a permissive and guiding environment for astrocytes. Accordingly, the functional properties of astrocytes measured via cell patch-clamp experiments reveal that PnNs do not alter the bioelectrical properties of resting astrocytes, thus setting the scene for the use of PnN-loaded nanofibres as bioconductive platforms for interfacing astrocytes and controlling their bioelectrical properties.


Pharmacokinetics of imipenem and cilastatin in patients with cystic fibrosis.
The pharmacokinetics of imipenem, a new carbapenem antibiotic, and cilastatin, a metabolic inhibitor, were evaluated in 17 patients with cystic fibrosis. Imipenem and cilastatin were combined in a ratio of 1:1 in the infusion solution, and patients intravenously received 30, 60, or 90 mg of imipenem per kg of body weight per day, divided into four equal doses. Pharmacokinetic evaluation after the first dose and again under steady-state conditions revealed biodisposition characteristics which were similar and independent of the daily dose administered. Cilastatin concentrations in serum paralleled those of imipenem. A linear relationship between dose and area under the serum concentration-time curve for both compounds was observed, suggesting a first-order pharmacokinetic process. A total of 50 and 78% of the doses of imipenem and cilastatin, respectively, were recovered unchanged in the urine. The renal clearances of imipenem and cilastatin averaged 54 and 88%, respectively, of the serum clearance. These data suggest that an extrarenal mechanism may be involved in the overall elimination of imipenem. No patient experienced any clinical or biochemical abnormalities during drug therapy.


Bite force and electromyograpy during maximum unilateral and bilateral clenching.
Maximum voluntary bite force has often been studied as an indicator of the functional state of the masticatory system. Bilateral, as well as unilateral, methods have been used to determine bite force. Only a few studies have compared the outcomes of both methods. The aim of this study was to measure bite force and jaw-muscle activity during bilateral as well as unilateral maximum clenching in a large number of healthy subjects, so that the results could be compared. In a group of 81 dentate subjects we observed an average bilateral bite force of 569 N. The average unilateral bite force was significantly lower, being 430 N (right) and 429 N (left). Masseter and anterior temporal muscle activities were also significantly lower during unilateral clenching as compared with bilateral clenching. The masseter muscles showed no difference in activity between the ipsilateral side and the contralateral side during unilateral clenching. In contrast, the activity of the anterior temporal muscle on the ipsilateral side was significantly higher than on the contralateral side. Thus, the change in the forces acting on the jaw during unilateral clenching compared with bilateral clenching leads to a different response in the temporal muscles than in the masseter muscles.


Mental health-related structural stigma and discrimination in health and social policies in Nepal: A scoping review and synthesis.
National policies can be used to reveal structural stigma and discrimination in relation to mental health. This review assesses how structural stigma and discrimination are manifested in the policies and legislations of Government of Nepal. Scoping review methodology was followed to review policy documents (acts of parliament, legislation, policies, strategies, guidelines and official directives) drafted or amended after 2010. Eighty-nine policies were identified related to health, social welfare, development and regulations which were relevant to people with psychosocial and mental disabilities or have addressed the mental health agendas. Several critical policy failings and gaps are revealed, such as the use of stigmatizing language (e.g., 'insane' or 'lunatic'), inconsistencies within and between policies, deviation from international protocols defining legal capacity and consent, lack of inclusion of the mental health agenda in larger development policies and lack of cost-effective interventions and identification of financing mechanisms. Provisions for people living with mental health conditions included adequate standard of living; attaining standard mental health; the right to exercise legal capacity, liberty and security; freedom from torture or discrimination; and right to live independently. However, other policies contradicted these rights, such as prohibiting marriage, candidacy for and retention of positions of authority and vulnerability to imprisonment. Mental health-related structural stigma and discrimination in Nepal can be identified through the use of discriminator language and provisions in the policies. The structural stigma and discrimination may be addressed through revision of the discriminating policies, integrating the mental health agenda into larger national and provincial policies, and streamlining policies to comply with national and international protocols.


Intra-abdominal polyphenotypic tumor.
The presence of t(11;22)(q24;q12) is often considered diagnostic of Ewing sarcoma and peripheral primitive neuroectodermal tumor (pPNET). We report a case of a polyphenotypic tumor that possessed this translocation as detected by reverse transcriptase polymerase chain reaction (RT-PCR). This tumor was positive for vimentin, desmin, low-molecular-weight keratin, neuron-specific enolase, S-100 protein, and CD57 by immunohistochemistry. Of note, the tumor was negative for MIC2. The tumor had double-minute chromosomes with > 100 copies of the MDM2 gene. Thus, the presence of the t(11;22)(q24;q12) translocation should not be considered diagnostic of Ewing sarcoma and pPNET in the absence of supporting histologic evidence such as positive staining for MIC2. The presence of this translocation in Ewing sarcoma and pPNET has been taken as evidence that these two tumors are related. Rather than extending this relationship to include some polyphenotypic tumors, other tumors may acquire this genetic change during tumor progression. Treatment regimens for tumors may be better based on phenotype rather than genotype when these two profiles are seemingly in conflict.


THE PRESENCE OF RENIN ACTIVITY IN BLOOD VESSEL WALLS.
The preparation, of an extract of hog blood vessels and organs containing renin activity has been described. The extract hydrolyzes natural or synthetic renin substrate to form a vasopressor material. This reaction could not be distinguished from the hydrolysis of renin substrates by kidney renin. The activation energy, effect of substrate concentration on velocity, inhibition by antirenin, and pH optimum of the extract and of kidney renin are the same. Renin activity is found in the adventitia and media of the aorta, the liver and, to a lesser extent, in other vascular organs.


Characterization of monoclonal antibodies directed against erythrocytic stage antigens of Plasmodium berghei.
Monoclonal antibodies recognizing various facets of the malaria parasite Plasmodium berghei and of the infected erythrocyte were obtained after generation of hybridomas between spleen cells from immunized mice and myeloma cells. The monoclonal antibodies were characterized by enzyme-linked immunosorbent assay, indirect immunofluorescence, immunoprecipitation of [35S]methionine-labeled proteins and immunoblotting. The most readily identified antigen was a parasite surface-associated protein of 230 kDa which is similar to the polymorphic schizont antigen described in a number of malarial species. In addition, three distinct antigens of 13, 31 and 120 kDa, which are external to the parasite, but within the infected erythrocyte were identified.


Genetic Variants of Cytochrome b-245, Alpha Polypeptide Gene and Premature Acute Myocardial Infarction Risk in an Iranian Population.
Oxidative stress induced by superoxide anion plays critical roles in the pathogenesis of coronary artery disease (CAD) and hence acute myocardial infarction (AMI). The major source of superoxide production in vascular smooth muscle and endothelial cells is the NADPH oxidase complex. An essential component of this complex is p22phox, that is encoded by the cytochrome b-245, alpha polypeptide (CYBA) gene. The aim of this study was to investigate the association of CYBA variants (rs1049255 and rs4673) and premature acute myocardial infarction risk in an Iranian population. The study population consisted of 158 patients under the age of 50 years, with a diagnosis of premature AMI, and 168 age-matched controls with normal coronary angiograms. Genotyping of the polymorphisms was performed by the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). There was no association between the genotypes and allele frequencies of rs4673 polymorphism and premature acute myocardial infarction (P>0.05). A significant statistical association was observed between the genotypes distribution of rs1049255 polymorphism and AMI risk (P=0.037). Furthermore, the distribution of AA+AG/GG genotypes was found to be statistically significant between the two groups (P=0.011). Our findings indicated that rs1049255 but not rs4673 polymorphism is associated with premature AMI.


Effects of different extents of pulley release on tendon excursion efficiency and tendon moment arms.
To compare the excursion efficiency and moment arms of flexor digitorum superficialis (FDS) and profundus (FDP) among different conditions of pulley integrity related to trigger finger treatment, cadaveric fingers were first tested with an intact pulley system, and then the first (A1) and second (A2) annular pulleys were released gradually from the proximal to distal part. Linear position sensors and a motion capture system were used to measure the tendon excursion and joint rotation simultaneously. The tendon excursion efficiency was defined as the range of motion of the involved joints per unit of tendon excursion, and the tendon moment arm was determined by the slope of the linear fitting result of tendon excursion versus metacarpophalangeal (MCP) joint rotation. No significant differences were found between the release of the A1 pulley and the release extending to half the proximal part of the A2 pulley in the FDP excursion efficiency and the moment arms of FDS and FDP with respect to the MCP joint. These results imply that the release could extend to half the proximal A2 pulley, if necessary, without significantly decreasing the FDP excursion efficiency and increasing the moment arms of FDS and FDP with respect to the MCP joint.


Mechanism of PhosphoThreonine/Serine Recognition and Specificity for Modular Domains from All-atom Molecular Dynamics.
Phosphopeptide-binding domains mediate many vital cellular processes such as signal transduction and protein recognition. We studied three well-known domains important for signal transduction: BRCT repeats, WW domain and forkhead-associated (FHA) domain. The first two recognize both phosphothreonine (pThr) and phosphoserine (pSer) residues, but FHA has high specificity for pThr residues. Here we used molecular dynamics (MD) simulations to reveal how FHA exclusively chooses pThr and how BRCT and WW recognize both pThr/pSer. The work also investigated the energies and thermodynamic information of intermolecular interactions. Simulations carried out included wide-type and mutated systems. Through analysis of MD simulations, we found that the conserved His residue defines dual loops feature of the FHA domain, which creates a small cavity reserved for only the methyl group of pThr. These well-organized loop interactions directly response to the pThr binding selectivity, while single loop (the 2nd phosphobinding site of FHA) or in combination with α-helix (BRCT repeats) or β-sheet (WW domain) fail to differentiate pThr/pSer. Understanding the domain pre-organizations constructed by conserved residues and the driving force of domain-phosphopeptide recognition provides structural insight into pThr specific binding, which also helps in engineering proteins and designing peptide inhibitors.


Selective transcriptional profiling and data analysis strategies for expression quantitative trait loci mapping in outbred F2 populations.
Genetic analysis of transcriptional profiling experiments is emerging as a promising approach for unraveling genes and pathways that underlie variation of complex biological traits. However, these genetical genomics approaches are currently limited by the high cost of microarrays. We studied five different strategies to optimally select subsets of individuals for transcriptional profiling, including (1) maximizing genetic dissimilarity between selected individuals, (2) maximizing the number of recombination events in selected individuals, (3) selecting phenotypic extremes within inferred genotypes of a previously identified quantitative trait locus (QTL), (4) purely random selection, and (5) profiling animals with the highest and lowest phenotypic values within each family-gender subclass. A simulation study was conducted on the basis of a linkage map and marker genotypes were derived from data on chromosome 6 for 510 F2 animals from an existing pig resource population and on a simulated biallelic QTL with pleiotropic effects on performance and gene expression traits. Bivariate analyses were conducted for selected subset sample sizes of 80, 160, and 240 individuals under three different correlation scenarios between the two traits. The genetic dissimilarity and phenotypic extremes within genotype methods had the smallest mean square error on QTL effects and maximum sensitivity on QTL detection, thereby outperforming all other selection strategies, particularly at the smallest proportion of samples selected for gene expression profiling (80/510).


High Heterogeneity of Escherichia coli Sequence Types Harbouring ESBL/AmpC Genes on IncI1 Plasmids in the Colombian Poultry Chain.
Escherichia coli producing ESBL/AmpC enzymes are unwanted in animal production chains as they may pose a risk to human and animal health. Molecular characterization of plasmids and strains carrying genes that encode these enzymes is essential to understand their local and global spread. To investigate the diversity of genes, plasmids and strains in ESBL/AmpC-producing E. coli from the Colombian poultry chain isolated within the Colombian Integrated Program for Antimicrobial Resistance Surveillance (Coipars). A total of 541 non-clinical E. coli strains from epidemiologically independent samples and randomly isolated between 2008 and 2013 within the Coipars program were tested for antimicrobial susceptibility. Poultry isolates resistant to cefotaxime (MIC ≥ 4 mg/L) were screened for ESBL/AmpC genes including blaCTX-M, blaSHV, blaTEM, blaCMY and blaOXA. Plasmid and strain characterization was performed for a selection of the ESBL/AmpC-producing isolates. Plasmids were purified and transformed into E. coli DH10B cells or transferred by conjugation to E. coli W3110. When applicable, PCR Based Replicon Typing (PBRT), plasmid Multi Locus Sequence Typing (pMLST), plasmid Double Locus Sequence Typing (pDLST) and/or plasmid Replicon Sequence Typing (pRST) was performed on resulting transformants and conjugants. Multi Locus Sequence Typing (MLST) was used for strain characterization. In total, 132 of 541 isolates were resistant to cefotaxime and 122 were found to carry ESBL/AmpC genes. Ninety-two harboured blaCMY-2 (75%), fourteen blaSHV-12 (11%), three blaSHV-5 (2%), five blaCTX-M-2 (4%), one blaCTX-M-15 (1%), one blaCTX-M-8 (1%), four a combination of blaCMY-2 and blaSHV-12 (4%) and two a combination of blaCMY-2 and blaSHV-5 (2%). A selection of 39 ESBL/AmpC-producing isolates was characterized at the plasmid and strain level. ESBL/AmpC genes from 36 isolates were transferable by transformation or conjugation of which 22 were located on IncI1 plasmids. These IncI1 plasmids harboured predominantly blaCMY-2 (16/22), and to a lesser extend blaSHV-12 (5/22) and blaCTX-M-8 (1/22). Other plasmid families associated with ESBL/AmpC-genes were IncK (4/33), IncHI2 (3/33), IncA/C (2/33), IncΒ/O (1/33) and a non-typeable replicon (1/33). Subtyping of IncI1 and IncHI2 demonstrated IncI1/ST12 was predominantly associated with blaCMY-2 (12/16) and IncHI2/ST7 with blaCTX-M-2 (2/3). Finally, 31 different STs were detected among the 39 selected isolates. Resistance to extended spectrum cephalosporins in E. coli from Colombian poultry is mainly caused by blaCMY-2 and blaSHV-12. The high diversity of strain Sequence Types and the dissemination of homogeneous IncI1/ST12 plasmids suggest that spread of the resistance is mainly mediated by horizontal gene transfer.


Developing a predictive tool for psychological well-being among Chinese adolescents in the presence of missing data.
Multi-dimensional behavioral rating scales like the CBCL and YSR are available for diagnosing psychosocial maladjustment in adolescents, but these are unsuitable for large-scale usage since they are time-consuming and their many sensitive questions often lead to missing data. This research applies multiple imputation to tackle the effects of missing data in order to develop a simple questionnaire-based predictive instrument for psychosocial maladjustment. Questionnaires from 2919 Chinese sixth graders in 21 schools were collected, but 86% of the students were missing one or more of the variables for analysis. Fifteen (10 training, 5 validation) samples were imputed using multivariate imputation chain equations. A ten-variable instrument was constructed by applying stepwise variable selection algorithms to the training samples, and its predictive performance was evaluated on the validation samples. The instrument had an AUC of 0.75 (95% CI: 0.73 to 0.78) and a calibration slope of 0.98 (95% CI: 0.86 to 1.09). The prevalence of psychosocial maladjustment was 18%. If a score of > 1 was used to define a negative test, then 80% of the students would be classified as negative. The resulting test had a diagnostic odds ratio of 5.64 (95% CI: 4.39 to 7.24), with negative and positive predictive values of 88% and 43%, and negative and positive likelihood ratios of 0.61 and 3.41, respectively. Multiple imputation together with internal validation provided a simple method for deriving a predictive instrument in the presence of missing data. The instrument's high negative predictive value implies that in populations with similar prevalences of psychosocial maladjustment test-negative students can be confidently excluded as being normal, thus saving 80% of the resources for confirmatory psychological testing.


Development of a robust method for isolation of shiga toxin-positive Escherichia coli (STEC) from fecal, plant, soil and water samples from a leafy greens production region in California.
During a 2.5-year survey of 33 farms and ranches in a major leafy greens production region in California, 13,650 produce, soil, livestock, wildlife, and water samples were tested for Shiga toxin (stx)-producing Escherichia coli (STEC). Overall, 357 and 1,912 samples were positive for E. coli O157:H7 (2.6%) or non-O157 STEC (14.0%), respectively. Isolates differentiated by O-typing ELISA and multilocus variable number tandem repeat analysis (MLVA) resulted in 697 O157:H7 and 3,256 non-O157 STEC isolates saved for further analysis. Cattle (7.1%), feral swine (4.7%), sediment (4.4%), and water (3.3%) samples were positive for E. coli O157:H7; 7/32 birds, 2/145 coyotes, 3/88 samples from elk also were positive. Non-O157 STEC were at approximately 5-fold higher incidence compared to O157 STEC: cattle (37.9%), feral swine (21.4%), birds (2.4%), small mammals (3.5%), deer or elk (8.3%), water (14.0%), sediment (12.3%), produce (0.3%) and soil adjacent to produce (0.6%). stx1, stx2 and stx1/stx2 genes were detected in 63%, 74% and 35% of STEC isolates, respectively. Subtilase, intimin and hemolysin genes were present in 28%, 25% and 79% of non-O157 STEC, respectively; 23% were of the "Top 6″ O-types. The initial method was modified twice during the study revealing evidence of culture bias based on differences in virulence and O-antigen profiles. MLVA typing revealed a diverse collection of O157 and non-O157 STEC strains isolated from multiple locations and sources and O157 STEC strains matching outbreak strains. These results emphasize the importance of multiple approaches for isolation of non-O157 STEC, that livestock and wildlife are common sources of potentially virulent STEC, and evidence of STEC persistence and movement in a leafy greens production environment.


Prevalence and correlates of vascular disease at ultrasound examination in patients on hemodialysis.
Patients on hemodialysis present an increased risk of cardiovascular death. Intimal media thickness (IMT) and presence of arterial calcifications are well-known risk factors for cardiovascular death in hemodialysis patients. To assess the prevalence of IMT and arterial calcifications in HD patients and to correlate image findings with clinical and laboratory data. Cross-sectional study involving 75 patients on dialysis for >12 months. Patients underwent B-mode ultrasound scan (US) for determination of IMT of the distal third of the common carotid arteries. Arterial calcifications were assessed by US of carotids, femoral and tibial arteries, and labeled positive if calcification was found in any arterial site. Patients were 52±13 years old, 57% were males and 16% were diabetics. IMT > 0.9 mm was found in 57% of cases and arterial calcifications at US in 48%. Aging (decades) and smoking were associated with both increased IMT (adjusted odds ratio [aOR] = 3.4, p < 0.001; aOR = 4.4, p = 0.045, respectively) and presence of vascular calcifications (aOR = 3.0, p < 0.001; aOR = 6.8, p = 0.011, respectively). High intact parathyroid hormone levels (per each 100 pg/ml) were significantly associated with increased IMT (aOR = 1.7, p = 0.021), but not with vascular calcification. In contrast, Diabetes and time on dialysis (years) were significant determinants for calcifications at US (aOR = 15.0, p = 0.009; aOR = 1.39, p = 0.020), but not for increased IMT. Increased IMT and calcifications at US are common findings in hemodialysis patients. Aging and smoking are consistent determinants for both image alterations. Parathyroid hormone elevation is associated with increased IMT. Diabetes and time on dialysis substantially increase the risk for arterial calcification.


Pancreas allotransplants in patients with a previous total pancreatectomy for chronic pancreatitis.
A total pancreatectomy is the last resort in the treatment of chronic pancreatitis because it results in complete endocrine and exocrine pancreatic insufficiency. More than 50% of total pancreatectomy patients experience severe glucose control problems, which cause up to 50% of late deaths. Between June 1, 1986, and May 15, 2007, we performed 26 pancreas allotransplants (18 primary, 8 retransplants) in 18 patients who had previously undergone a total pancreatectomy for chronic pancreatitis. All patients had a history of labile diabetes mellitus with hypoglycemic unawareness; secondary diabetic complications developed in 12. The median time interval from the total pancreatectomy to the pancreas allotransplant was 5 years (range 9 months to 22 years). Of the 26 transplants, 6 were performed in the cyclosporine (CSA) era, 15 in the tacrolimus (TAC) era, and 5 in the calcineurin inhibitor (CNI)-free era. Patient survival rates at 1 and 3 years in both the CSA and TAC eras were 100% and 100%; in the CNI-free era, at 1 year, the survival rate was 40%. Pancreas graft survival rates in the CSA era were 67% and 50% at 1 and 3 years, respectively; in the TAC era, 73% and 51%, respectively; and in the CNI-free era, at 1 year, 40% (p=0.13). The mean number of rejection episodes in the CSA era was 2.1; in the TAC era, 1.4; and in the CNI-free era, 0.6. Our series of pancreas allotransplants in patients with a previous total pancreatectomy for chronic pancreatitis showed that pancreas graft survival rates of more than 70% can be achieved with TAC-based immunosuppression; pancreas transplants can successfully treat both endocrine and exocrine insufficiency; and sequential pancreas allotransplants should be considered a treatment option in patients with pancreatectomy-induced brittle diabetes mellitus or with progression of secondary complications of diabetes mellitus.


The nutritional composition of British bread: London area study.
Samples of white and brown bread, both sliced and unsliced, and of wheatgerm breads and wholemeal bread were purchased in London and analysed for a wide range of nutrients. Available carbohydrate, dietary fibre, fatty acids, riboflavin, vitamin B6, and iodine were determined in bulked samples of each type of bread and, in addition, every loaf was analysed for moisture, fat, protein, phosphorus, chloride, sodium, potassium, calcium, magnesium, iron, zinc, copper, manganese, thiamin, nicotinic acid, and free and total folic acid, in order to provide an estimate the variability within each kind of bread. Virtually all the constituents that were measured in the individual loaves showed inter-loaf variation--particularly moisture contents and the levels of calcium, copper and folic acid. Unsliced loaves were more variable than sliced loaves but when expressed on a dry matter basis there were no significant differences in the nutrient levels in sliced and unsliced breads. Wheatgerm breads were relatively more homogenous products but wholemeal loaves were very unhomogenous reflecting the difficulty of accurately identifying unwrapped wholemeal loaves in retail outlets. Some differences from previously published values for all breads were found for dietary fibre, iron, thiamin and vitamin B6. Experience of sampling at retail outlets and analysis provided by this study will be integrated into the design of a planned study of breads throughout Britain.


ECG Signal-Enabled Automatic Diagnosis Technology of Heart Failure.
Usually, heart failure occurs when heart-related diseases are developed and continue to deteriorate veins and arteries. Heart failure is the final stage of heart disease, and it has become an important medical problem, particularly among the aging population. In medical diagnosis and treatment, the examination of heart failure contains various indicators such as electrocardiogram. It is one of the relatively common ways to collect heart failure or attack related information and is also used as a reference indicator for doctors. Electrocardiogram indicates the potential activity of patient's heart and directly reflects the changes in it. In this paper, a deep learning-based diagnosis system is presented for the early detection of heart failure particularly in elderly patients. For this purpose, we have used two datasets, Physio-Bank and MIMIC-III, which are publicly available, to extract ECG signals and thoroughly examine heart failure. Initially, a heart failure diagnosis model which is based on attention convolutional neural network (CBAM-CNN) is proposed to automatically extract features. Additionally, attention module adaptively learns the characteristics of local features and efficiently extracts the complex features of the ECG signal to perform classification diagnosis. To verify the exceptional performance of the proposed network model, various experiments were carried out in the realistic environment of hospitals. Influence of signal preprocessing on the performance of model is also discussed. These results show that the proposed CBAM-CNN model performance is better for both classifications of ECG signals. Likewise, the CBAM-CNN model is sensitive to noise, and its accuracy is effectively improved as soon as signal is refined.


Long term outcome of benign childhood epilepsy with centrotemporal spikes: Dutch Study of Epilepsy in Childhood.
To determine long-term outcome in a cohort of children with newly diagnosed benign childhood epilepsy with centrotemporal spikes (BECTS). 29 children with BECTS were included in the Dutch Study of Epilepsy in Childhood. Each child was followed for 5 years, and subsequently contacted 12-17 years after enrolment to complete a structured questionnaire. Twenty children had typical BECTS, nine had atypical BECTS (age at onset <4 years, developmental delay or learning difficulties at inclusion, other seizure types, atypical EEG abnormalities). Mean age at onset of epilepsy was 8.0 years with slight male preponderance. Most common seizure-types before enrolment were generalized tonic-clonic seizures (GTCS) and simple partial seizures; in 86% of the children seizures occurred during sleep. After 12-17 years, 96% had a terminal remission (TR(F)) of more than 5 years and 89% of more than 10 years. Mean duration of epilepsy was 2.7 years; mean age at reaching TR(F) was 10.6 years. Many children (63%) had experienced one or more (secondary) GTCS. Antiepileptic drugs were used by 79% of the children with a mean duration of 3.0 years. None of the children seemed to have developed learning problems or an arrest of cognitive development during follow-up. No significant differences were observed in patient characteristics or outcome between children with typical BECTS and children with atypical BECTS. All children in our cohort, both those with typical and atypical BECTS, had a very good prognosis with high remission rates after 12-17 years. None of the predictive factors for disease course and outcome observed in earlier studies (other seizure types, age at onset, multiple seizures at onset) were prognostic in our cohort.


Differential gene regulation by a synthetic vitamin D receptor ligand and active vitamin D in human cells.
Vitamin D (VD) exerts a wide variety of biological functions including calcemic activity. VD nutritional status is closely associated with the onset and development of chronic diseases. To develop a VD analog with the desired VD activity but without calcemic activity, we screened synthetic VDR antagonists. We identified 1α,25-dihydroxyvitamin D3-26-23-lactams (DLAM)-2a-d (DLAM-2s) as nuclear vitamin D receptor (VDR) ligands in a competitive VDR binding assay for 1α,25(OH)2 vitamin D3 (1α,25(OH)2D3), and DLAM-2s showed an antagonistic effect on 1α,25(OH)2 D3-induced cell differentiation in HL60 cells. In a luciferase reporter assay in which human VDR was exogenously expressed in cultured COS-1 cells, DLAM-2s acted as transcriptional antagonists. Consistently, DLAM-2s had an antagonistic effect on the 1α,25(OH)2D3-induced expression of a known VD target gene [Cytochrome P450 24A1 (CYP24A1)], and VDR bound DLAM-2s was recruited to an endogenous VD response element in chromatin in human keratinocytes (HaCaT cells) endogenously expressing VDR. In an ATAC-seq assay, the effects of 1α,25(OH)2 D3 and DLAM-2b on chromatin reorganization were undetectable in HaCaT cells, while the effect of an androgen receptor (AR) antagonist (bicalutamide) was confirmed in prostate cancer cells (LNCaP) expressing endogenous AR. However, whole genome analysis using RNA-seq and ATAC (Assay for Transposase Accessible Chromatin)-seq revealed differential gene expression profiles regulated by DLAM-2b versus 1α,25(OH)2D3. The upregulated and downregulated genes only partially overlapped between cells treated with 1α,25(OH)2D3 and those treated with DLAM-2b. Thus, the present findings illustrate a novel VDR ligand with gene regulatory activity differing from that of 1α,25(OH)2D3.


A Toxoplasma gondii 10 kDa in vitro excretory secretory antigen reactive with human IgM and IgA antibodies.
Toxoplasmosis can cause serious disease in immunocompromised patients and to congenitally infected foetuses. Appropriate laboratory investigations in potential cases of acute Toxoplasma infection are important. Excretory secretory antigen (ESA) is immunogenic during both human and experimental infections, therefore is considered as a good candidate for investigation into new infection markers. In this study, ESA was prepared from in vitro cultures of Toxoplasma gondii to identify T. gondii ESA antigenic component(s) that is/are most reactive with serum samples from probable acute cases of toxoplasmosis. Serum samples were obtained from several categories of individuals with the following Toxoplasma serology: Group I: IgM+ IgG+ (low IgG avidity) or IgM+ IgG- from sera of patients who had clinical query of toxoplasmosis (n=35). Group II: IgM- IgG+ (high IgG avidity) from chronically infected individuals (n=30). Group III: normal/healthy individuals with anti-Toxoplasma IgMIgG- (n=20). Group IV: individuals with other infections who had anti-Toxoplasma IgM- IgG- (n=10). The ESA was subjected to SDS-PAGE, followed by Western blot analysis using the above sera and probed with peroxidase conjugated anti-human IgM and IgA antibodies. The blots were then developed using chemiluminescence substrate. The selected antigenic band was excised from the gel after two dimensional electrophoresis and sent for mass spectrometry analysis using MALDI TOF-TOF. The most promising antigenic band was a 10 kDa protein which showed sensitivity of 80% in both IgM and IgA blots, and specificity of 96.7% with sera from other infections and healthy controls. The two best identifications for the 10 kDa band were ubiquitin (ribosomal protein CEP52 fusion protein) and polyubiquitin.


Pentamidine inhibits catalytic activity of group I intron Ca.LSU by altering RNA folding.
The antimicrobial agent pentamidine inhibits the self-splicing of the group I intron Ca.LSU from the transcripts of the 26S rRNA gene of Candida albicans, but the mechanism of pentamidine inhibition is not clear. We show that preincubation of the ribozyme with pentamidine enhances the inhibitory effect of the drug and alters the folding of the ribozyme in a pattern varying with drug concentration. Pentamidine at 25 microM prevents formation of the catalytically active F band conformation of the precursor RNA and alters the ribonuclease T1 cleavage pattern of Ca.LSU RNA. The effects on cleavage suggest that pentamidine mainly binds to specific sites in or near asymmetric loops of helices P2 and P2.1 on the ribozyme, as well as to the tetraloop of P9.2 and the loosely paired helix P9, resulting in an altered structure of helix P7, which contains the active site. Positively charged molecules antagonize pentamidine inhibition of catalysis and relieve the drug effect on ribozyme folding, suggesting that pentamidine binds to a magnesium binding site(s) of the ribozyme to exert its inhibitory effect.


Arsenic trioxide inhibits p-glycoprotein expression in multidrug-resistant human leukemia cells that overexpress the MDR1 gene.
To investigate the effects of arsenic trioxide (As(2)O(3)) on the apoptosis and p-glycoprotein (P-gp) expression of multidrug-resistant human leukemia cells. Human multidrug-resistant leukemia cell line K562/ADM overexpressing the MDR1 gene, was used as the target cells. The cell proliferating activity was assessed using the MTT colorimetric assay. Cytomorphology was investigated under light, confocal and electron microscopes. DNA fragmentation was examined using agarose gel electrophoresis, while p-gp expression, cell cycle status and sub-G1 cells were determined using flow cytometry. Zero point five to 20 micromol/L As(2)O(3) inhibited the proliferation of K562/ADM cells, and K562/ADM cells were more sensitive to As(2)O(3) than the parental K562 cells. As(2)O(3)-induced apoptosis of K562/ADM cells was determined by the observance of typical morphological changes and the appearance of DNA ladder and sub-G1 cell populations. As(2)O(3) significantly inhibited the P-gp expression of K562/ADM cells, and synergistically enhanced the sensitivity of the drug-resistant cells to adriamycin. As(2)O(3) induces growth-inhibition and apoptosis, down-regulates P-gp expression and exerts a synergistic effect in combination with adriamycin in multidrug-resistant leukemia cells.


Comparison of the effectiveness between power toothbrushes and manual toothbrushes for oral health: a systematic review and meta-analysis.
Objective: Power toothbrushes is considered an effective tool for maintaining oral health; however, its efficacy as compared to manual toothbrushes is still not completely clarified. This article aims to evaluate the efficacy of power toothbrushes compared with the manual toothbrushes in terms of plaque, gingivitis and bleeding reduction.Methods: An electronic search was performed on PUBMED, Web of Science, Wiley and Research Gate. Studies comparing the effectiveness of plaque, gingivitis and bleeding reduction between power and manual toothbrushes were included. Results and effect sizes analysis are presented as standard mean difference (SMD), and subgroup analysis stratified by mode of action of the power toothbrush was performed. Study quality and risk of bias were assessed using the Cochrane assessment tool.Results: A total of 21 randomized clinical studies were included. Power toothbrushes were significantly more effective in reducing plaque index (26 trials: SMD = 0.86, 95% CI: 0.58 to 1.14, I2 = 91.5%, p < .0001), gingival index (14 trials: SMD = 0.47, 95% CI: 0.12 to 0.82, I2 = 88.7%, p < .0001), and bleeding index (11 trials: SMD = 0.92, 95% CI: 0.43 to 1.40, I2 = 91.8%, p < .0001) compared with the manual toothbrushes, except that there was no significant differences between the oscillating-rotating toothbrushes and manual toothbrushes regarding gingivitis reduction (7 trials: SMD = 0.07, 95% CI: -0.20 to 0.33, I2 = 57.2%, p = .03).Conclusions: Power toothbrushes is more effective in reducing dental plaque, gingivitis and bleeding compared with the manual toothbrush.


Organic removal activity in biofilm and suspended biomass fractions of MBBR systems.
The moving bed biofilm reactor (MBBR) wastewater treatment process is usually designed based on the assumption that all activity in the process occurs in the biofilm on the MBBR carriers, although there is always some active biomass in the bulk liquid due to biofilm sloughing and, sometimes, free-growing bacteria. In this study the removal of organic matter is evaluated in laboratory-scale MBBR reactors under varying load, hydraulic retention time (HRT), oxygen concentration and volumetric filling degree of carriers in order to determine the heterotrophic activity in the different fractions of the MBBR biomass. The results showed that the heterotrophic conversions in an MBBR can show the same type of diffusion limited dependency on oxygen as nitrification, even for easily degradable substrates such as acetate. The contribution to the removal from the suspended biomass is shown to vary depending on HRT, as the amount of suspended solids changes. The developed method in this report is a useful tool for determining heterotrophic activity in the separate fractions of biomass in MBBRs.


Novel Macrocyclic Peptidomimetics Targeting the Polo-Box Domain of Polo-Like Kinase 1.
The polo-box domain (PBD) of Plk1 is a promising target for cancer therapeutics. We designed and synthesized novel phosphorylated macrocyclic peptidomimetics targeting PBD based on acyclic phosphopeptide PMQSpTPL. The inhibitory activities of 16e on Plk1-PBD is >30-fold higher than those of PMQSpTPL. Both 16a and 16e possess excellent selectivity for Plk1-PBD over Plk2/3-PBD. Analysis of the cocrystal structure of Plk1-PBD in complex with 16a reveals that the 3-(trifluoromethyl)benzoyl group in 16a interacts with Arg516 through a π-stacking interaction. This π-stacking interaction, which has not been reported previously, provides insight into the design of novel and potent Plk1-PBD inhibitors. Furthermore, 16h, a PEGlyated macrocyclic phosphopeptide derivative, induces Plk1 delocalization and mitotic failure in HeLa cells. Also, the number of phospho-H3-positive cells in a zebrafish embryo increases in proportion to the amount of 16a. Collectively, the novel macrocyclic peptidomimetics should serve as valuable templates for the design of potent and novel Plk1-PBD inhibitors.


Efficacy of Dapagliflozin According to Heart Rate: A Patient-Level Pooled Analysis of DAPA-HF and DELIVER.
Although elevated resting heart rate (HR) is associated with a higher risk of cardiovascular events in patients with heart failure with reduced ejection fraction in sinus rhythm (SR), the relationship between HR and outcomes among patients with heart failure with mildly reduced ejection fraction/heart failure with preserved ejection fraction and in those with atrial fibrillation (AF) is uncertain. The aims of this study were to examine the association between baseline HR and outcomes across the range of left ventricular ejection fraction, in patients with and without AF, and evaluate the effect of dapagliflozin according to HR. A patient-level pooled analysis of the DAPA-HF (Dapagliflozin and Prevention of Adverse Outcomes in Heart Failure; heart failure with reduced ejection fraction) and DELIVER (Dapagliflozin Evaluation to Improve the Lives of Patients With Preserved Ejection Fraction Heart Failure trial; heart failure with mildly reduced ejection fraction/heart failure with preserved ejection fraction) trials. The primary outcome of each was the composite of worsening heart failure or cardiovascular death. Among patients with SR (n=6401, 64%), the rate of the primary outcome was higher in those with higher HR: 16.8 versus 7.7 per 100 person-years for ≥80 bpm versus <60 bpm. The relationship between HR and risk was steeper in heart failure with reduced ejection fraction versus heart failure with mildly reduced ejection fraction/heart failure with preserved ejection fraction. HR was not associated with outcomes in patients in AF for either heart failure phenotype. The benefit of dapagliflozin on the primary outcome was consistent across the HR range in both SR (Pinteraction=0.28) and AF (Pinteraction=0.56), for example, for SR <60 bpm, hazard ratio for dapagliflozin versus placebo 0.72 (95% CI, 0.55-0.95); 60 to 69 bpm, 0.78 (0.63-0.97); 70 to 79 bpm, 0.73 (0.59-0.91); ≥80 bpm, 0.77 (0.61-0.97). The benefit was consistent across HR range in both heart failure with reduced ejection fraction and heart failure with mildly reduced ejection fraction/heart failure with preserved ejection fraction. The risk of worsening heart failure or cardiovascular death increased with increasing baseline HR among patients in SR, but this association was not seen among patients in AF, irrespective of left ventricular ejection fraction. The benefit of dapagliflozin was consistent across HR range, irrespective of left ventricular ejection fraction or rhythm. URL: https://www. gov; Unique identifier: NCT03036124 and NCT03619213.


Free-radical reaction in biological systems.
Free radicals are a highly reactive chemical species which differ from all other species in possessing an unpaired electron. Although free-radical activity in living systems is energetically improbable, recent work suggests that a number of important biological processes depend on it. In contrast to enzymic metabolism these free-radical processes tend to be non-cyclic, irreversible, non-homoeostatic, and energetically wasteful. For the organism as a whole they may nevertheless have considerable survival value. Free radicals are too short-lived to be demonstrable in tissues; but methodological advances are leading to the detection and measurement of characteristic free-radical reaction products. The findings suggest that free-radical activity is not only a potential cause but also a common, perhaps invariable, consequence of cell damage. It is possible that some of the secondary products of free-radical reactions may help to regulate the body's local and systemic response to injury.


Atomically resolved orientational ordering of C60 molecules on epitaxial graphene on Cu(111).
A detailed understanding of interactions between molecules and graphene is one of the key issues for tailoring the properties of graphene-based molecular devices, because the electronic and structural properties of molecular layers on surfaces are determined by intermolecular and molecule-substrate interactions. Here, we present the atomically resolved experimental measurements of the self-assembled fullerene molecules on single-layer graphene on Cu(111). Fullerene molecules form a (4 × 4) superstructure on graphene/Cu(111), revealing only single molecular orientation. We can resolve the exact adsorption site and the configuration of fullerene by means of low-temperature scanning tunnelling microscopy (LT-STM) and density functional theory (DFT) calculations. The adsorption orientation can be explained in terms of the competition between intermolecular interactions and molecule-substrate interactions, where strong Coulomb interactions among the fullerenes determine the in-plane orientation of the fullerene. Our results provide important implications for developing carbon-based organic devices using a graphene template in the future.


Sleep states and behavior patterns in preterm and fullterm infants.
The sleep states and state-related behaviors of 10 preterm infants were naturalistically observed at 37 and 41 weeks CA; 20 fullterm infants were also observed at 41 weeks CA. Respiration was recorded in synchrony with the behavioral observations. At 41 weeks CA the preterm infants seemed more mature than the fullterm infants on some measures, and less mature on others. These results indicate global differences in state organization, although neither the causes nor the consequences of these differences are currently known.


Teaching professionalism - Why, What and How.
Due to changes in the delivery of health care and in society, medicine became aware of serious threats to its professionalism. Beginning in the mid-1990s it was agreed that if professionalism was to survive, an important step would be to teach it explicitly to students, residents, and practicing physicians. This has become a requirement for medical schools and training programs in many countries. There are several challenges in teaching professionalism. The first challenge is to agree on the definition to be used in imparting knowledge of the subjects to students and faculty. The second is to develop means of encouraging students to consistently demonstrate the behaviors characteristic of a professional - essentially to develop a professional identity. Teaching of professionalism must be both explicit and implicit. The cognitive base consisting of definitions and -attributes and medicine's social contract with society must be taught and evaluated explicitly. Of even more -importance, there must be an emphasis on experiential learning and reflection on personal experience. The general principles, which can be helpful to an institution or program of teaching professionalism, are presented, along with the experience of McGill University, an institution which has established a comprehensive program on the teaching of professionalism.


Graphene-Enabled, Spatially Controlled Electroporation of Adherent Cells for Live-Cell Super-resolution Microscopy.
The incorporation of exogenous molecules into live cells is essential for both biological research and therapeutic applications. In particular, for the emerging field of super-resolution microscopy of live mammalian cells, it remains a challenge to deliver tailored, often cell-impermeable, fluorescent probes into live cells for target labeling. Here, utilizing the outstanding mechanical, electrical, and optical properties of graphene, we report a facile approach that enables both high-throughput delivery of fluorescent probes into adherent mammalian cells and in situ super-resolution microscopy on the same device. Approximately 90% delivery efficiencies are achieved for free dyes and dye-tagged affinity probes, short peptides, and whole antibodies, thus enabling high-quality super-resolution microscopy. Moreover, we demonstrate good spatiotemporal controls, which, in combination with the ready patternability of graphene, allow for the spatially selective delivery of two different probes for cells at different locations on the same substrate.


Does comorbid substance use disorder impair recovery from major depression with SSRI treatment? An analysis of the STAR*D level one treatment outcomes.
Many patients with major depressive disorder (MDD) present with concurrent substance use disorders (SUDs), which has been thought to impair their response to antidepressants. Clinicians often delay antidepressant treatment until sustained sobriety has been established. Unfortunately, these comorbid subjects are typically excluded from depression treatment trials, leaving a gap in understanding the treatment outcomes. In the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study, 2876 adult outpatients diagnosed with nonpsychotic MDD were prospectively treated with the selective serotonin-reuptake inhibitor (SSRI), citalopram, and returned for at least one post-baseline visit. Participants with SUD (29%) and without SUD (71%) were compared in regard to baseline clinical and sociodemographic features and treatment response. The group with MDD and SUD was further subdivided into those with alcohol only, drug only, and both alcohol and drug use. Despite clear sociodemographic and clinical differences, there were no significant differences between groups in the time to achieve response or rates of response to citalopram; however, those who endorsed both alcohol and drug use had significantly reduced rates of remission and significantly increased times to reach remission compared to the MDD group without SUD. In addition, subjects with MDD and SUD had higher risk of psychiatric serious adverse events (3.3% vs. 1.5%) and hospitalization (2.8% vs. 1.2%). The results indicate that first-line treatment with citalopram in depressed patients with alcohol or drug use respond as well as those without SUD. More intensive treatment is most likely needed for MDD patients with both drug and alcohol use disorders.


Suppression of PKC-α attenuates TNF-α-evoked cerebral barrier breakdown via regulations of MMP-2 and plasminogen-plasmin system.
Ischaemic stroke, accompanied by neuroinflammation, impairs blood-brain barrier integrity through a complex mechanism involving both protein kinase C (PKC) and urokinase. Using an in vitro model of human blood-brain barrier (BBB) composed of brain microvascular endothelial cells (HBMEC) and astrocytes, this study assessed the putative roles of these elements in BBB damage evoked by enhanced availability of pro-inflammatory cytokine, TNF-α. Treatment of HBMEC with TNF-α significantly increased the mRNA and protein expressions of all plasminogen-plasmin system (PPS) components, namely tissue plasminogen activator, urokinase, urokinase plasminogen activator receptor and plasminogen activator inhibitor-1 and also the activities of urokinase, total PKC and extracellular MMP-2. Inhibition of urokinase by amiloride abated the effects of TNF-α on BBB integrity and MMP-2 activity without affecting that of total PKC. Conversely, pharmacological inhibition of conventional PKC isoforms dramatically suppressed TNF-α-induced overactivation of urokinase. Knockdown of PKC-α gene via specific siRNA in HBMEC suppressed the stimulatory effects of TNF-α on protein expression of all PPS components, MMP-2 activity, DNA fragmentation rates and pro-apoptotic caspase-3/7 activities. Establishment of co-cultures with BMEC transfected with PKC-α siRNA attenuated the disruptive effects of TNF-α on BBB integrity and function. This was partly due to elevations observed in expression of a tight junction protein, claudin-5 and partly to prevention of stress fibre formation. In conclusion, specific inhibition of PKC-α in cerebral conditions associated with exaggerated release of pro-inflammatory cytokines, notably TNF-α may be of considerable therapeutic value and help maintain endothelial cell viability, appropriate cytoskeletal structure and basement membrane.


Use of proteins to minimize the physical aging of EUDRAGIT sustained release films.
The objective of this study was to investigate the influence of two proteins, albumin and type B gelatin, on the physical aging of EUDRAGIT RS 30 D and RL 30 D coated theophylline pellets. The physicomechanical properties of sprayed films, thermal properties of cast films, influence of proteins on the zeta potential and particle size of the dispersion, and the release of proteins from cast films under simulated dissolution conditions were investigated. The release rate of theophylline decreased significantly over time from pellets coated with an acrylic dispersion containing 10% albumin when there was no acidification of the acrylic dispersion; however, when pellets were coated with an acidified EUDRAGIT/albumin dispersion, the theophylline release rate was stable for dosage forms stored in the absence of humidity. The drug release rate was faster for pellets coated with acrylic dispersions containing 10% gelatin compared to the albumin-containing formulations. When sprayed films were stored at 40 degrees C/75% RH, the water vapor permeability decreased significantly for both EUDRAGIT films and those containing EUDRAGIT and albumin; however, there was no significant change in this parameter when 10% gelatin was present. Albumin was released from the acrylic films when the pH of the dissolution media was below the isoelectric point of the protein while no quantitative release of gelatin was observed in pH 1.2 or 7.4 media. The effect of gelatin to prevent the decrease in drug release rate was due to stabilization in water vapor permeability of the film. Acidification of the polymeric dispersion resulted in electrostatic repulsive forces between albumin and the acrylic polymer, which stabilized the drug release rate when the dosage forms were stored in aluminum induction sealed containers at both 40 degrees C/75% RH and 25 degrees C/60% RH.


Frontal and temporal dopamine release during working memory and attention tasks in healthy humans: a positron emission tomography study using the high-affinity dopamine D2 receptor ligand [11C]FLB 457.
Experimental studies on animals have shown that dopamine is a key neurotransmitter in the regulation of working memory (WM) functions in the prefrontal cortex. In humans, blood flow studies show prefrontal involvement in WM functions, but direct evidence for the involvement of the dopaminergic system in WM is lacking. Using positron emission tomography with a recently developed high-affinity dopamine D2 receptor tracer, [11C]FLB 457, we explored frontal, temporal, and parietal D2 receptor availability in 12 healthy volunteers while they were performing verbal WM and sustained attention tasks. During the performance of both tasks, reduced D2 receptor availability was observed in the left ventral anterior cingulate, suggesting an attention or arousal-related increase in dopamine release during these tasks. Compared with the sustained attention task, the verbal WM task reduced D2 receptor availability in the ventrolateral frontal cortex bilaterally and in the left medial temporal structures (amygdala, hippocampus), suggesting that dopamine release in these regions might have a specific role in WM. In addition, correlation analyses indicated that increased dopamine release in the right ventrolateral frontal cortex and the left ventral anterior cingulate during the WM task was associated with faster and more stable WM performance, respectively. Our results indicate that regionally specific components of the frontotemporal dopaminergic network are functionally involved in WM performance in humans.


Inappropriate activation of androgen receptor by relaxin via beta-catenin pathway.
We have previously demonstrated that human H2-relaxin can mediate androgen-independent growth of LNCaP through a mechanism that involves the activation of the androgen receptor (AR) signaling pathway. The goal of the current study is to elucidate the mechanism(s) by which H2-relaxin causes activation of the AR pathway. Our data indicate that there is cross-talk between AR and components of the Wnt signaling pathway. Addition of H2-relaxin to LNCaP cells resulted in increased phosphorylation of protein kinase B (Akt) and inhibitory phosphorylation of glycogen synthase kinase-3beta (GSK-3beta) with subsequent cytoplasmic accumulation of beta-catenin. Immunoprecipitation and immunocytochemical studies demonstrated that the stabilized beta-catenin formed a complex with AR, which was then translocated into the nucleus. Chromatin immunoprecipitation analysis determined that the AR/beta-catenin complex binds to the proximal region of the prostate-specific antigen promoter. Inhibition of the phosphatidylinositol 3-kinase (PI3K)/Akt pathway, using LY294002, prevented both H2-relaxin-mediated phosphorylation of Akt and GSK-3beta and translocation of beta-catenin/AR into the nucleus. Knockdown of beta-catenin levels using a beta-catenin-specific small interfering RNA inhibited H2-relaxin-induced AR activity. The combined data demonstrate that PI3K/Akt and components of the Wnt pathway can facilitate H2-relaxin-mediated activation of the AR pathway.


Drinking Location and Pregaming as Predictors of Alcohol Intoxication Among Mandated College Students.
Both drinking location and pregaming have been associated with heavy alcohol use among college students, yet the manner by which they uniquely contribute to alcohol intoxication remains unclear. The current study examined the unique utility of drinking location and pregaming in predicting alcohol intoxication among college students who violated campus alcohol policy. Between 2011 and 2012, mandated college students who reported drinking prior to their referral events (N = 212, 41% female, 80% White, Mage = 19.4 y) completed a computerized assessment of drinking location and related behaviors as part of larger research trial. Chi-squared statistics, t-tests, one-way analyses of covariance, and regression were used to examine study aims. Participants were most likely (44%) to report drinking in off-campus housing prior to the referral event, and approximately half (47%) reported pregaming. Alcohol intoxication on the night of the referral event differed significantly as a function of both drinking location and pregaming, but pregaming did not moderate the association between drinking location and alcohol intoxication among mandated students. Female birth sex, pregaming, and drinking at either fraternities or off-campus housing predicted greater levels of alcohol intoxication on the night of the referral incident, while drinking in a residence hall/dorm predicted lower intoxication. Drinking location and pregaming are distinct predictors of alcohol intoxication among mandated college students. Future interventions may benefit from targeting both where and how college students consume alcohol.


Microfluidic Electroporation Coupling Pulses of Nanoseconds and Milliseconds to Facilitate Rapid Uptake and Enhanced Expression of DNA in Cell Therapy.
Standard electroporation with pulses in milliseconds has been used as an effective tool to deliver drugs or genetic probes into cells, while irreversible electroporation with nanosecond pulses is explored to alter intracellular activities for pulse-induced apoptosis. A combination treatment, long nanosecond pulses followed by standard millisecond pulses, is adopted in this work to help facilitate DNA plasmids to cross both cell plasma membrane and nuclear membrane quickly to promote the transgene expression level and kinetics in both adherent and suspension cells. Nanosecond pulses with 400-800 ns duration are found effective on disrupting nuclear membrane to advance nuclear delivery of plasmid DNA. The additional microfluidic operation further helps suppress the negative impacts such as Joule heating and gas bubble evolution from common nanosecond pulse treatment that lead to high toxicity and/or ineffective transfection. Having appropriate order and little delay between the two types of treatment with different pulse duration is critical to guarantee the effectiveness: 2 folds or higher transfection efficiency enhancement and rapid transgene expression kinetics of GFP plasmids at no compromise of cell viability. The implementation of this new electroporation approach may benefit many biology studies and clinical practice that needs efficient delivery of exogenous probes.


A microplate assay to measure classical and alternative complement activity.
We developed and validated a kinetic microplate hemolytic assay (HA) to quantify classical and alternative complement activity in a single dilution of human plasma or serum. The assay is based on monitoring hemolysis of sensitized sheep (or uncoated rabbit) red blood cells by means of a 96-well microplate reader. The activity of the calibrator was evaluated by reference to 200 healthy adults. The conversion of 50% hemolysis time into a percentage of activity was obtained using a calibration curve plotted daily. The linearity of the assay as well as interference (by hemolysis, bilrubinemia and lipemia) was assessed for classical pathway (CP). The within-day and the between-day precision was satisfactory regarding the performance of commercially available liposome immunoassay (LIA) and ELISA. Patients with hereditary or acquired complement deficiencies were detected (activity was measured <30%). We also provided a reference range obtained from 200 blood donors. The agreement of CP evaluated on samples from 48 patients was 94% with LIA and 87.5% with ELISA. The sensitivity of our assay was better than that of LIA, and the cost was lower than either LIA or ELISA. In addition, this assay was less time consuming than previously reported HAs. This assay allows the simultaneous measurement of 36 samples in duplicate per run of a 96-well plate. The use of a daily calibration curve allows standardization of the method and leads to good reproducibility. The same technique was also adapted for the quantification of alternative pathway (AP) activity.


Machine learning and tubercular drug target recognition.
Tuberculosis (TB) remains to be a global major public-health threat, causing millions of deaths each year. A major difficulty in dealing with TB is that the causative bacterium, Mycobacterium tuberculosis, can persist in host tissue for a long period of time even after treatment. Mycobacterial persistence has become a central research focus for developing next-generation TB drugs. Latest genomic technology has enabled a high-throughput approach for identifying potential TB drug targets. Each gene product can be screened for its uniqueness to the TB metabolism, host-pathogen discrimination, essentiality for survival, and potential for chemical binding, among other properties. However, the exhaustive search for useful drug targets over the entire genome would not be productive as expected in practice. On the other hand, the problem can be formulated as pattern recognition or inductive learning and tackled with rule-based or statistically based learning algorithms. Here we review the perspective that combines machine learning and genomics for drug discovery in tuberculosis.


Denoising of Quality Scores for Boosted Inference and Reduced Storage.
Massive amounts of sequencing data are being generated thanks to advances in sequencing technology and a dramatic drop in the sequencing cost. Much of the raw data are comprised of nucleotides and the corresponding quality scores that indicate their reliability. The latter are more difficult to compress and are themselves noisy. Lossless and lossy compression of the quality scores has recently been proposed to alleviate the storage costs, but reducing the noise in the quality scores has remained largely unexplored. This raw data is processed in order to identify variants; these genetic variants are used in important applications, such as medical decision making. Thus improving the performance of the variant calling by reducing the noise contained in the quality scores is important. We propose a denoising scheme that reduces the noise of the quality scores and we demonstrate improved inference with this denoised data. Specifically, we show that replacing the quality scores with those generated by the proposed denoiser results in more accurate variant calling in general. Moreover, a consequence of the denoising is that the entropy of the produced quality scores is smaller, and thus significant compression can be achieved with respect to lossless compression of the original quality scores. We expect our results to provide a baseline for future research in denoising of quality scores. The code used in this work as well as a Supplement with all the results are available at http://web.stanford.edu/~iochoa/DCCdenoiser_CodeAndSupplement.zip.


Concentration Effect of Reducing Agents on Green Synthesis of Gold Nanoparticles: Size, Morphology, and Growth Mechanism.
Under various concentration conditions of reducing agents during the green synthesis of gold nanoparticles (AuNPs), we obtain the various geometry (morphology and size) of AuNPs that play a crucial role in their catalytic properties. Through both theoretical and experimental approaches, we studied the relationship between the concentration of reducing agent (caffeic acid) and the geometry of AuNPs. As the concentration of caffeic acid increases, the sizes of AuNPs were decreased due to the adsorption and stabilizing effect of oxidized caffeic acids (OXCAs). Thus, it turns out that optimal concentration exists for the desired geometry of AuNPs. Furthermore, we investigated the growth mechanism for the green synthesis of AuNPs. As the caffeic acid is added and adsorbed on the surface of AuNPs, the aggregation mechanism and surface free energy are changed and consequently resulted in the AuNPs of various geometry.


Genetic admixture and cardiovascular disease risk in postmenopausal Hispanic women.
Hispanics are a heterogeneous population with differences in the prevalence of cardiovascular disease (CVD) and its related risk factors among ethnic sub-groups. This study evaluated the association of genetic admixture and CVD in self-identified Hispanic women from the Women's Health Initiative (WHI). Data came from the WHI Observational Study and the Clinical Trial Components conducted among postmenopausal women. The CVD outcomes included coronary heart disease (CHD) and stroke. The proportions of European (EUR), sub-Saharan African (AFR), and Amerindian (AMI) admixture were estimated using 92 ancestry-informative markers. Cox regression models were used to assess the relationship between genetic admixture and CVD adjusting for age, lifestyle risk factors, known risk factors, and neighborhood socioeconomic status. Among 5195 participants EUR ancestry was associated with a lower CHD risk after adjusting for age (HR 0.41, p = 0.02), and in the fully adjusted model (HR 0.40, p = 0.03). AFR ancestry was associated with a higher CHD risk after adjusting for age (HR 2.91, p = 0.03), but it only showed a trend in in the fully adjusted model (HR 2.46, p = 0.10). AMI ancestry was not statistically significantly associated with CHD and none of the genetic admixture proportions were statistically significantly associated with stroke (p > 0.05). EUR ancestry was associated with a lower risk of CHD in Hispanic women. This highlights the need to account for genetic admixture in future CVD studies to consider different heritage groups to understand the role that genetic, neighborhood socioeconomic status, and environmental factors contribute to CVD health disparities in Hispanic women.


Potential for Genetic Improvement of the Main Slaughter Yields in Common Carp With in vivo Morphological Predictors.
Common carp is a major aquaculture species worldwide, commonly sold alive but also as processed headless carcass or filets. However, recording of processing yields is impossible on live breeding candidates, and alternatives for genetic improvement are either sib selection based on slaughtered fish, or indirect selection on correlated traits recorded in vivo. Morphological predictors that can be measured on live fish and that correlate with real slaughter yields hence remain a possible alternative. To quantify the power of morphological predictors for genetic improvement of yields, we estimated genetic parameters of slaughter yields and various predictors in 3-year-old common carp reared communally under semi-intensive pond conditions. The experimental stock was established by a partial factorial design of 20 dams and 40 sires, and 1553 progenies were assigned to their parents using 12 microsatellites. Slaughter yields were highly heritable (h2 = 0.46 for headless carcass yield, 0.50 for filet yield) and strongly genetically correlated with each other (rg = 0.96). To create morphological predictors, external (phenotypes, 2D digitization) and internal measurements (ultrasound imagery) were recorded and combined by multiple linear regression to predict slaughter yields. The accuracy of the phenotypic prediction was high for headless carcass yield (R2 = 0.63) and intermediate for filet yield (R2 = 0.49). Interestingly, heritability of predicted slaughter yields (0.48-0.63) was higher than that of the real yields to predict, and had high genetic correlations with the real yields (rg = 0.84-0.88). In addition, both predicted yields were highly phenotypically and genetically correlated with each other (0.95 for both), suggesting that using predicted headless carcass yield in a breeding program would be a good way to also improve filet yield. Besides, two individual predictors (P1 and P2) included in the prediction models and two simple internal measurements (E4 and E23) exhibited intermediate to high heritability estimates (h2 = 0.34 - 0.72) and significant genetic correlations to the slaughter yields (rg = |0.39 - 0.83|). The results show that there is a solid potential for genetic improvement of slaughter yields by selecting for predictor traits recorded on live breeding candidates of common carp.


Effects of feeding polydextrose on faecal characteristics, microbiota and fermentative end products in healthy adult dogs.
Polydextrose is a potential prebiotic, but has not been well tested in dogs. Thus, the objective of the present study was to determine the effects of polydextrose on faecal characteristics, microbial populations and fermentative end products in healthy adult dogs. A total of eight adult hound dogs (3.5 (sem 0.5) years; 20 (sem 0.5) kg) were randomly allotted to one of four test diets containing the following concentrations of polydextrose: (1) 0 % (control); (2) 0.5 %; (3) 1.0 %; or (4) 1.5 %. A Latin square design was used, with each treatment period lasting 14 d (days 0-10 adaptation; days 11-14 fresh and total faecal collection). All dogs were fed to maintain body weight. Data were evaluated for linear and quadratic effects using SAS software. Although apparent total tract DM digestibility was unaffected, total tract crude protein digestibility tended to decrease (P < 0.10) linearly with increasing dietary polydextrose concentrations. Fresh faecal DM percentage tended to decrease (P < 0.10) linearly, while faecal scores increased (P < 0.05; looser stools) with increasing dietary concentrations of polydextrose. Faecal acetate, propionate and total SCFA concentrations increased (P < 0.05) linearly with increased dietary polydextrose. Faecal pH decreased (P < 0.05) linearly with increasing polydextrose. Faecal indole tended to decrease (P < 0.10) linearly with increasing polydextrose, but other faecal protein catabolites were not changed. Faecal Clostridium perfringens linearly decreased (P < 0.05) with increasing dietary polydextrose concentrations, but Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. were not affected. Based on the present results, polydextrose appears to act as a highly fermentable fibre, but requires further research to test its potential as a prebiotic in dogs.


Amino acid pools and metabolism during the cell division cycle of arginine-grown Candida utilis.
Synchronous cultures obtained by isopycnic density gradient centrifugation are used to investigate amino acid metabolism during the cell division cycle of the food yeast Candida utilis. Isotopic labeling experiments demonstrate that the rates of uptake and catabolism of arginine, the sole source of nitrogen, double abruptly during the first half of the cycle, while the cells undergo bud expansion. This is accompanied by a doubling in rate of amino acid biosynthesis, and an accumulation of amino acids. The accumulation probably occurs within the storage pools of the vacuoles. Amino acids derived from protein degradation contribute little to this accumulation. For the remainder of the cell cycle, during cell separation and until the next bud initiation, the rates of uptake and catabolism of arginine and amino acid biosynthesis remain constant. Despite the abrupt doubling in the rate of formation of amino acid pools, their rate of utilization for macromolecular synthesis increases steadily throughout the cycle. The significance of this temporal organization of nitrogen source uptake and amino acid metabolism during the cell division cycle is discussed.


Relevance of quality improvement methods to surgical practice: prospective assessment of carotid endarterectomy.
Continuous quality improvement methods are increasingly being applied to health care systems, yet demonstration of outcome and cost benefits for surgical patients remains sparse. We used continuous quality improvement principles to specifically identify potential opportunities to reduce patient charges for carotid endarterectomy in our academic vascular surgery practice without compromising results. The targeted opportunities included: 1) limitation of laboratory examination, 2) selective cardiac stress testing, 3) discharge on 1st postoperative day, and 4) substitution of outpatient carotid duplex imaging for inpatient angiography. After 1 year, reductions in the average patient charge ($7700 versus $13,900, P < 0.001) and increases in payment/charge ratio (1.2 versus 0.8; P < 0.001) were observed. These changes were primarily due to a reduction in length of stay (2.2 versus 5.7 days; P < 0.001). No significant difference in patient morbidity occurred. Reductions in charges occurred within the targeted areas of laboratory (-77%), cardiac testing (-73%), hospital room (-60%), and radiology (-81%) utilization. Attention to the four factors identified by continuous quality improvement methods significantly reduced total patient charges without detrimental effects on patient outcome.


Attitudes and opinions on corporal punishment among urban Israeli Jews.
Corporal punishment of children constitutes a human rights violation and has mental health consequences, yet it is frequently practiced in many countries. The objectives of this study were to identify attitudes and opinions with regard to corporal punishment among urban Jewish Israelis. Corporal punishment was defined as the means to discipline children by the use of physical force. A short battery of questions exploring attitudes and opinions on the use of corporal punishment of children was added to a periodic survey conducted by a private polling organization. The sample included 1,000 urban Jewish Israeli community residents. Most respondents did not endorse corporal punishment. However, when confronted with a question tapping the readiness of adults to comply with a ruling by the Supreme Court that prohibits the use of corporal punishment at home, most respondents expressed skepticism that the public will comply. Forty percent thought that most parents would agree to undergo training in the use of alternative means of discipline. Fifty-seven percent experienced physical punishment when growing up, a variable which was associated with all responses favoring its use as a means of discipline. The results of the study provide the bases for public health action, including training programs on alternative methods of discipline. Further research targeting other population groups not yet explored is needed.


A metal-mediated base pair that discriminates between the canonical pyrimidine nucleobases.
A nucleoside analogue comprising the ligand 1H-imidazo[4,5-f][1,10]phenanthroline (P) was applied to develop a molecular beacon capable of discriminating the canonical nucleobases cytosine and thymine. The beacon is based on the formation of a stable Ag+-mediated base pair between P and cytosine, whereas the presence of Ag+ strongly destabilizes nucleic acids comprising an artificial base pair between P and thymine. Metal-mediated base pair formation was investigated by temperature-dependent UV spectroscopy and CD spectroscopy and complemented by extensive DFT calculations. The molecular beacon significantly extends the application spectrum of nucleic acids with metal-mediated base pairs. It is of potential use in the detection of single-nucleotide polymorphisms.


Aging and the kidney.
A host of abnormalities of renal structure and function accompanies advancing age. An appreciation of methodologic considerations, including population selection, that might confound the assessment of the effects of aging on renal function has prompted a recent reappraisal. Earlier studies assessed the effects of aging by utilizing cross-sectional studies and institutionalized elderly subjects, with their attendant drawbacks. Recent longitudinal studies have utilized appropriate patient cohorts, selected for lock of renal disease, including potential kidney transplant donors. These studies indicate that the morphological and functional changes of aging tend to be less marked than previously thought. The common denominator of these functional changes is a diminution in renal reserve, along with constraints on the kidney's ability to respond appropriately to challenges of either excesses or deficits. Although these alterations are unlikely to be of major clinical consequence under everyday conditions, they attain clinical significance when residual renal function is challenged by the superimposition of an acute illness. Finally, it should be emphasized that elderly patients frequently suffer from comorbid conditions, such as hypertension and heart disease, that may be additive to the changes of aging, thereby amplifying these abnormalities.


Bacterial adhesion in oligotrophic habitats.
Oligotrophic and copiotrophic bacteria can act as primary colonizers of surfaces in low nutrient habitats. Adhesion in copiotrophs provides an opportunity to change from a starvation-survival existence in the aqueous phase to an active growth mode at the solid surface. Healthy daughter cells are released from the surface to colonize other surfaces or to resume starvation-survival in the aqueous phase. Non-adhesive copiotrophs scavenge nutrients at surfaces, a fact seldom recognized in studies on the partitioning of bacteria between particulates and the aqueous phase.


Adrenal cavernous hemangioma: a case report with review of the literature.
Adrenal cavernous hemangioma is a rare type of tumor that is usually diagnosed post-operatively. There have only been approximately 63 cases reported in the literature to date. We report a case of adrenal cavernous hemangioma in a 27-year-old pregnant woman. The mass was discovered on ultrasonography when she visited a gastroenterologist for vague epigastric discomfort and vomiting. The laboratory tests were within normal limits and did not show any features suggestive of adrenal endocrinologic dysfunction. Computed Tomography (CT) revealed a well-defined 7.8 × 7.8 oval mass in the right adrenal gland with speckled calcifications. The mass was removed by transabdominal laparoscopic surgery. Strong positive immunostaining for CD31 and CD34 with weakly positive staining for podoplanin/D2-40 confirmed the diagnosis of cavernous hemangioma. We reviewed 52 case reports of adrenal cavernous hemangioma in an attempt to identify tumor characteristics. More than half of the patients reviewed showed a heterogeneous internal structure of the mass with peripheral patchy enhancement on CT. They also showed focal or speckled calcifications either on X-ray or CT. Nevertheless, many of these characteristics overlap with the imaging phenotypes of other common diseases of the adrenal gland and therefore do not seem to provide definite evidence for differential diagnosis. Laparoscopic approach is a feasible and safe modality to remove adrenal cavernous hemangiomas because they seem to form a rigid fibrotic capsule; hence the risk of bleeding due to surgical manipulation is relatively low.


Elucidating the Anomalous Binding Enhancement of Isoquinoline Boronic Acid for Sialic Acid Under Acidic Conditions: Expanding Biorecognition Beyond Vicinal Diols.
A zwitterionic heterocyclic boronic acid based on 4-isoquinolineboronic acid (IQBA) exhibits the highest reported binding affinity for sialic acid or N-acetylneuraminic acid (Neu5Ac, K=5390±190 m-1 ) through the formation of a cyclic boronate ester complex under acidic conditions (pH 3). This anomalous pH-dependent binding enhancement does not occur with common neutral saccharides (e.g., glucose, fructose, sorbitiol), because it is mediated via selective complexation to a α-hydroxycarboxylate moiety forming a stable ion pair and ternary complex with Neu5Ac in phosphate buffer. IQBA expands biorecognition beyond classical vicinal diols under neutral or alkaline buffer conditions, which enables the direct analysis of Neu5Ac by native fluorescence with sub-micromolar detection limits.


Allele-specific PCR amplification due to sequence identity between a PCR primer and an amplicon: is direct sequencing so reliable?
PCR-direct sequencing (DS) is thought to be a very reliable method of determining DNA sequence and genotyping. Under certain conditions, however, DS can generate inaccurate results. Here we report a case of erroneous DS, in which a single nucleotide polymorphism (SNP) in the human PAX9 gene was mistyped due to allele-dependent PCR amplification. Examination of the amplified region showed that the 5' eight bases of one of the PCR primers were identical to the eight bases of the reverse strand downstream of the SNP, and the ninth base matched one of the alleles. Altering the primer so that it matched the other allele reversed the allele-specific inhibition. Reducing the base-pairing abolished the inhibition. Thus, the SNP was responsible for the difference in annealing efficacy of the primer and was therefore critical for the allele dependency. The allele-specific inhibition presented here can occur with any PCR primer sequence that encompasses a site that is polymorphic in the gene sequence. This phenomenon needs to be considered as a possibility when interpreting results from all PCR-based experiments. Sequence similarity between PCR primers and internal amplified regions should be considered for all methods for mutation detection and genotyping using PCR.