Water Relations in Pulvini from Samanea saman: II. Effects of Excision of Motor Tissues.
Pulvinar motor tissues of Samanea saman are often excised for in vitro studies of ion transport. Because ion transport may be regulated in part by hydrostatic pressure (P), this study explores how P and water potential (Psi) change when motor tissues are excised. Water potential (Psi) of excised extensor and flexor tissues was measured by the Chardakov method and compared with Psi measurements made on extensor and flexor tissues of intact pulvini (HL Gorton 1987 Plant Physiol 83: 945-950). Psi values for excised extensor and flexor tissues were always substantially more negative than for the same tissues in intact pulvini. Extensor tissues excised from open pulvini had slightly more negative Psi than excised flexor tissues, and the opposite was true for closed pulvini. Extensor and flexor tissues elongate immediately when excised from open or closed pulvini, suggesting that in intact pulvini they are constrained from elongating by the nonextensible vascular core. In addition, both tissues in both open and closed pulvini are under compression imposed by oppositely positioned motor tissue. Excision relieves constraint and compression, decreasing P, and thus decreasing Psi. This finding may explain, at least in part, the difference between Psi measurements on intact and excised motor tissues. Implications of these data for the planning and interpretation of in vitro experiments requiring excised strips of extensor and flexor tissues are discussed.


An Update in Epigenetics in Metabolic-Associated Fatty Liver Disease.
Metabolic-associated fatty liver disease (MAFLD) is characterized by hepatic steatosis accompanied by one of three features: overweight or obesity, T2DM, or lean or normal weight with evidence of metabolic dysregulation. It is distinguished by excessive fat accumulation in hepatocytes, and a decrease in the liver's ability to oxidize fats, the accumulation of ectopic fat, and the activation of proinflammatory pathways. Chronic damage will keep this pathophysiologic cycle active causing progression from hepatic steatosis to cirrhosis and eventually, hepatocarcinoma. Epigenetics affecting gene expression without altering DNA sequence allows us to study MAFLD pathophysiology from a different perspective, in which DNA methylation processes, histone modifications, and miRNAs expression have been closely associated with MAFLD progression. However, these considerations also faced us with the circumstance that modifying those epigenetics patterns might lead to MAFLD regression. Currently, epigenetics is an area of great interest because it could provide new insights in therapeutic targets and non-invasive biomarkers. This review comprises an update on the role of epigenetic patterns, as well as innovative therapeutic targets and biomarkers in MAFLD.


Nicotine does not affect vascularization but inhibits growth of freely transplanted ovarian follicles by inducing granulosa cell apoptosis.
There is clear support for an association between smoking and decreased female fecundity and fertility. Cigarette smoke appears to have adverse effects along a continuum of reproductive processes. We therefore studied the effect of nicotine on follicular growth and vascularization of freely transplanted ovarian follicles. We used the skinfold chamber model in Syrian golden hamsters, which allows the in vivo microscopy of follicular grafts. Animals were treated daily with nicotine at doses mimicking low-rate and high-rate smokers (0.2 and 1.0 mg/kg body weight subcutaneously). Saline-treated animals served as controls. To further evaluate the effect of nicotine on angiogenesis, an in vitro aortic ring assay was used. The re-vascularisation rate of follicles was similar in nicotine-treated animals and controls. During the 7 days after transplantation, nicotine further caused a dose-dependent inhibition of follicular growth. In contrast, the vascularized area and microvessel density were not affected by the nicotine exposure. In vitro aortic ring assays confirmed that nicotine does not influence sprouting and microvessel formation. However, immunohistochemistry for cleaved caspase-3 revealed a large extent of granulosa cell apoptosis within transplanted follicles of high-dose nicotine-treated animals. Nicotine as one toxic component of cigarette smoke does not affect vascularization, but adversely influences follicular growth by an increase in apoptotic cell death. As follicular growth is a crucial step in normal ovulation and fertilization, nicotine-induced cell apoptosis may represent one of the mechanisms underlying the well-established link between smoking and fertility disorders.


[HLA-G: an immunoregulatory non classical class I HLA molecule].
HLA-G is a particular non classical HLA class I molecule. Despite its tissue-restricted expression and low polymorphism, this molecule plays an important role in innate and adaptative immunity. The tolerogenic propriety of HLA-G makes it an immunomodulatory molecule acting in the early phases of conception, protecting fetal tissues from the maternal immune system. Immunomodulatory functions of HLA-G and the associations of this molecule with some pathological states are reported in this review. So, little amounts of soluble HLA-G or particular allelic expression of this molecule are associated with some pregnancy complications. HLA-G expression on tumor cells by preventing antitumor responses via a trogocytosis mechanism and regulatory T cells induction is associated with invasiveness and clinical evolution of some tumor types. HLA-G is also involved in the protection of the transplanted tissues from rejection. Revealing of new more functional homomultimeric isoforms of this molecule offers new insight in a better understanding of clinical and biological role of HLA-G.


Efficacy of the Herpes Simplex Virus 2 (HSV-2) Glycoprotein D/AS04 Vaccine against Genital HSV-2 and HSV-1 Infection and Disease in the Cotton Rat Sigmodon hispidus Model.
Subunit vaccines based on the herpes simplex virus 2 (HSV-2) glycoprotein D (gD-2) have been the major focus of HSV-2 vaccine development for the past 2 decades. Based on the promising data generated in the guinea pig model, a formulation containing truncated gD-2, aluminum salt, and MPL (gD/AS04) advanced to clinical trials. The results of these trials, however, were unexpected, as the vaccine protected against HSV-1 infection but not against HSV-2. To address this discrepancy, we developed a Depot medroxyprogesterone acetate (DMPA)-treated cotton rat Sigmodon hispidus model of HSV-2 and HSV-1 genital infection. The severity of HSV-1 genital herpes was less than that of HSV-2 genital herpes in cotton rats, and yet the model allowed for comparative evaluation of gD/AS04 immunogenicity and efficacy. Cotton rats were intramuscularly vaccinated using a prime boost strategy with gD/AS04 (Simplirix vaccine) or control vaccine formulation (hepatitis B vaccine FENDrix) and subsequently challenged intravaginally with HSV-2 or HSV-1. The gD/AS04 vaccine was immunogenic in cotton rats and induced serum IgG directed against gD-2 and serum HSV-2 neutralizing antibodies but failed to efficiently protect against HSV-2 disease or to decrease the HSV-2 viral load. However, gD/AS04 significantly reduced vaginal titers of HSV-1 and better protected animals against HSV-1 compared to HSV-2 genital disease. The latter finding is generally consistent with the clinical outcome of the Herpevac trial of Simplirix. Passive transfer of serum from gD/AS04-immunized cotton rats conferred stronger protection against HSV-1 genital disease. These findings suggest the need for alternative vaccine strategies and the identification of new correlates of protection. In spite of the high health burden of genital herpes, there is still no effective intervention against the disease. The significant gap in knowledge on genital herpes pathogenesis has been further highlighted by the recent failure of GSK HSV-2 vaccine Simplirix (gD/AS04) to protect humans against HSV-2 and the surprising finding that the vaccine protected against HSV-1 genital herpes instead. In this study, we report that gD/AS04 has higher efficacy against HSV-1 compared to HSV-2 genital herpes in the novel DMPA-synchronized cotton rat model of HSV-1 and HSV-2 infection. The findings help explain the results of the Simplirix trial.


New artificial electron donors for in vitro assay of nitrate reductase isolated from cultured tobacco cells and other organisms.
The capacity of bromphenol blue and its analogs to act as electron donors for measurement of in vitro nitrate reductase activity from tobacco cells (Nicotiana tabacum var Techné SP 25 strain) was determined. Competitive inhibition was demonstrated to occur between NADH, the natural electron donor, and bromphenol blue, the artificial electron donor, suggesting that both donors bind to a similar active site on the enzyme. NADH-dependent or bromphenol blue-dependent nitrate reductase activity was carried out by a similar molecular weight protein exhibiting similar antigenic sites. Following ammonium sulfate precipitation, sucrose density gradient and two chromatographic steps, nitrate reductase activity from tobacco cells was purified near homogeneity using bromphenol blue as an electron donor in the absence of measurable NADH-dependent activity. The enzyme is composed of two identical subunits of 83 kilodaltons < Momega < 94 kilodaltons.


Deep Surgical Site Infections after Open Radical Cystectomy and Urinary Diversion Significantly Increase Hospitalisation Time and Total Treatment Costs.
Deep surgical site infections (DSSI) usually require secondary treatments. The aim of this study was to compare the total length of hospitalisation (LOH), intensive care unit (ICU) duration, and total treatment costs in patients with DSSI versus without DSSI after open radical cystectomy (ORC) and urinary diversion. Prospective case-control study in a tertiary care hospital in patients after ORC with urinary diversion during April 2008 to July 2012. DSSI was defined based on Centers for Disease Control and Prevention criteria. Matched-pair analysis for patients with versus without DSSI was done in 1:2 ratios. Patients with superficial surgical site infections (SSI) were excluded from analysis. In total, 189 operations were performed. Thirty-eight patients (20.1%) developed SSI of which 28 patients (14.8%) had DSSI. Out of 28 patients, 27 (96.4%) were with DSSI and required surgical re-intervention. Due to insufficient matching criteria, 11 patients with DSSI were excluded from analyses. Consequently, 17 patients with DSSI were matched with 34 patients without DSSI. Significant differences were seen for median overall LOH (30 vs. 18 days, p < 0.001), median ICU duration (p = 0.024), and median overall treatment costs (€17,030 vs. €11,402, p = 0.011). DSSI significantly increases LOH (67%) and treatment costs (49%), adding up to a financial loss for the hospital of approximately €5,500 in patients with DSSI.


Hepatitis C virus antibody in Poland.
To assess the epidemiology of hepatitis C virus (HCV) in Poland, anti-HCV was studied in patients with acute and chronic non-A, non-B (NANB) hepatitis, in healthy adults, and in subjects at risk. Anti-HCV prevalence was 2% in 152 blood donors, 78% in 95 parenteral drug addicts, 21% in 112 alcoholics, and 86% in 42 patients with chronic NANB hepatitis. Among 34 prospectively followed patients with acute NANB hepatitis 17 (50%) developed anti-HCV. It seems that HCV infection is responsible for the majority of NANB hepatitis in Poland and is common in parenteral drug abusers and alcoholics.


Transcriptome meta-analysis of peripheral lymphomononuclear cells indicates that gestational diabetes is closer to type 1 diabetes than to type 2 diabetes mellitus.
We performed a meta-analysis of the transcription profiles of type 1, type 2 and gestational diabetes to evaluate similarities and dissimilarities among these diabetes types. cRNA samples obtained from peripheral blood lymphomononuclear cells (PBMC) of 56 diabetes mellitus patients (type 1 = 19; type 2 = 20; gestational = 17) were hybridized to the same whole human genome oligomicroarray platform, encompassing 44,000 transcripts. The GeneSpring software was used to perform analysis and hierarchical clustering, and the DAVID database was used for gene ontology. The gene expression profiles showed more similarity between gestational and type 1 diabetes rather than between type 2 and gestational diabetes, a finding that was not influenced by patient gender and age. The meta-analysis of the three types of diabetes disclosed 3,747 differentially and significantly expressed genes. A total of 486 genes were characteristic of gestational diabetes, 202 genes of type 1, and 651 genes of type 2 diabetes. 19 known genes were shared by type 1, type 2 and gestational diabetes, highlighting EGF, FAM46C, HBEGF, ID1, SH3BGRL2, VEPH1, and TMEM158 genes. The meta-analysis of PBMC transcription profiles characterized each type of diabetes revealing that gestational and type 1 diabetes were transcriptionally related.


Gastrointestinal surgery in cystic fibrosis: a 20-year review.
The purpose of this study was to evaluate outcomes of the surgical management for meconium ileus (MI) and Distal Intestinal Obstruction Syndrome (DIOS) in Cystic Fibrosis (CF). Children born between 1990 and 2010 were identified using a regional CF database. Retrospective case note analysis was performed. Outcome measures for MI were mortality, relaparotomy rate, length of stay (LOS), time on parental nutrition (TP), and time to full feeds (TFF). Outcome measures for DIOS were: age of onset, number of episodes, and need for laparotomy. Seventy-five of 376 neonates presented with MI. Fifty-four (92%) required laparotomy. Contrast enema decompression was attempted in nineteen. There were no post-operative deaths. Thirty-nine (72%) neonates with MI were managed with stomas. LOS was longer in those managed with stomas (p=0.001) and in complex MI (p=0.002). Thirty-five patients were treated for DIOS. Twenty-five patients were managed with gastrograffin. Ten patients underwent surgical management of DIOS. Overall, MI did not predispose to later development of DIOS. There was a significantly greater incidence of laparotomy for DIOS in children who had MI. The proportion of neonates with complex meconium ileus was high (49%) and may explain the infrequent utilisation of radiological decompression. Complex MI or management with stomas both significantly increase LOS. Re-laparotomy rate is high (22%) in MI irrespective of the type of management. DIOS is not a benign condition, particularly when the child has had previous abdominal surgery. Early referral to a surgical team is essential in these children.


Analytical validation of a practical molecular assay prognostic of survival in nonsquamous non-small cell lung cancer.
A molecular assay prognostic of survival in resected nonsquamous non-small cell lung cancer designed to meet the need for improved risk stratification in early-stage disease has recently been described. This assay measures the expression levels of 14 genes using RNA extracted from formalin-fixed, paraffin-embedded (FFPE) tissues. The assay underwent blinded clinical validation in 2 large international cohorts involving approximately 1500 patients; the analytical precision and reproducibility of this assay, however, have not yet been reported. For each of the 14 TaqMan quantitative polymerase chain reaction (PCR) primer and probe sets used in the molecular prognostic assay, the linear range, PCR efficiency, limits of blank, limits of quantitation, and quantitative bias were determined using serial dilutions of pooled RNA extracted from FFPE samples. The reproducibility of the entire molecular assay was determined by performing repeat testing of FFPE samples over multiple days. The linear range of individual quantitative TaqMan PCR primer and probe sets was between 2(10)- and 2(15)-fold input RNA. The median C(T) of the quantitative PCR primer and probe sets at 10 ng of input RNA was 24.3; the median efficiency was 91.2%. The median quantitative bias across all quantitative PCR primer and probe sets was 0.75% (range, 0.32% to 1.32%). In repeat testing, the mean SD of the risk score (scaled from 1 to 100) was 2.18, with a mean coefficient of variation of 0.08. The molecular prognostic assay presented in this study demonstrates high precision and reproducibility, validating its clinical utility as a reliable prognostic tool that can contribute to the management of patients with early-stage disease.


Interaction between trehalose and MTHase from Sulfolobus solfataricus studied by theoretical computation and site-directed mutagenesis.
Maltooligosyltrehalose trehalohydrolase (MTHase) catalyzes the release of trehalose by cleaving the α-1,4-glucosidic linkage next to the α-1,1-linked terminal disaccharide of maltooligosyltrehalose. Computer simulation using the hydrogen bond analysis, free energy decomposition, and computational alanine scanning were employed to investigate the interaction between maltooligosyltrehalose and the enzyme. The same residues that were chosen for theoretical investigation were also studied by site-directed mutagenesis and enzyme kinetic analysis. The importance of residues determined either experimentally or computed theoretically were in good accord with each other. It was found that residues Y155, D156, and W218 of subsites -2 and -3 of the enzyme might play an important role in interacting with the ligand. The theoretically constructed structure of the enzyme-ligand complex was further validated through an ab initio quantum chemical calculation using the Gaussian09 package. The activation energy computed from this latter study was very similar to those reported in literatures for the same type of hydrolysis reactions.


[Extraction of buildings three-dimensional information from high-resolution satellite imagery based on Barista software].
The demand for accurate and up-to-date spatial information of urban buildings is becoming more and more important for urban planning, environmental protection, and other vocations. Today's commercial high-resolution satellite imagery offers the potential to extract the three-dimensional information of urban buildings. This paper extracted the three-dimensional information of urban buildings from QuickBird imagery, and validated the precision of the extraction based on Barista software. It was shown that the extraction of three-dimensional information of the buildings from high-resolution satellite imagery based on Barista software had the advantages of low professional level demand, powerful universality, simple operation, and high precision. One pixel level of point positioning and height determination accuracy could be achieved if the digital elevation model (DEM) and sensor orientation model had higher precision and the off-Nadir View Angle was relatively perfect.


Deficiency of Bid protein reduces sepsis-induced apoptosis and inflammation, while improving septic survival.
Increased apoptotic cell death is believed to play a pathological role in patients with sepsis and experimental animals. Apoptosis can be induced by either a cell death receptor (extrinsic) or a mitochondrial (intrinsic) pathway. Bid, a proapoptotic member of the Bcl-2 family, is thought to mediate the cross talk between the extrinsic and intrinsic pathways of apoptosis; however, little is known about the action of Bid in the development of apoptosis and organ-specific tissue damage/cell death as seen in polymicrobial sepsis. Our results show that after the onset of sepsis, tBid (the active form of Bid) is significantly increased in mitochondrial fractions of the thymus, spleen, Peyer patches, and liver, and that Fas or FasL deficiency blocks Bid activation in various tissues after septic challenge. Increased Bid activation is correlated with increased active caspase-3, caspase-9, and apoptosis during sepsis. Bid-deficient mice exhibit significantly reduced apoptosis in the thymus, spleen, and Peyer patches compared with background mice after sepsis. Furthermore, Bid-deficient mice had significantly reduced systemic and local inflammatory cytokine levels and improved survival after sepsis. These data support not only the contribution of Bid to sepsis-induced apoptosis and the onset of septic morbidity/mortality, but also the existence of a bridge between extrinsic apoptotic signals, e.g., FasL:Fas, TNF:TNFR, and so on, and the intrinsic mitochondrial pathway via Bid-tBid activation during sepsis.


Validity of inspiratory and expiratory methods of measuring gas exchange with a computerized system.
The accuracy of a computerized metabolic system, using inspiratory and expiratory methods of measuring ventilation, was assessed in eight male subjects. Gas exchange was measured at rest and during five stages on a cycle ergometer. Pneumotachometers were placed on the inspired and expired side to measure inspired (VI) and expired ventilation (VE). The devices were connected to two systems sampling expired O(2) and CO(2) from a single mixing chamber. Simultaneously, the criterion (Douglas bag, or DB) method assessed VE and fractions of O(2) and CO(2) in expired gas (FE(O(2)) and FE(CO(2))) for subsequent calculation of O(2) uptake (VO(2)), CO(2) production (VCO(2)), and respiratory exchange ratio. Both systems accurately measured metabolic variables over a wide range of intensities. Though differences were found between the DB and computerized systems for FE(O(2)) (both inspired and expired systems), FE(CO(2)) (expired system only), and VO(2) (inspired system only), the differences were extremely small (FE(O(2)) = 0.0004, FE(CO(2)) = -0.0003, VO(2) = -0.018 l/min). Thus a computerized system, using inspiratory or expiratory configurations, permits extremely precise measurements to be made in a less time-consuming manner than the DB technique.


Immunolocalization of oviductin in endocytic compartments in the blastomeres of developing embryos in the golden hamster.
We have previously localized an antigen of oviductal origin in the zona pellucida of superovulated hamster ova. This antigen is a high-molecular-weight glycoprotein that is secreted by the nonciliated secretory cells of the oviduct and later is transferred to the zona pellucida of the oocyte during oviductal transit. This glycoprotein is rich in N-acetyl-D-galactosamine residues and has been designated Hamster Oviductin-1. In the present study, we have examined the intracellular localization of the oviductin in the blastomeres of developing embryos of the golden hamster. Seventeen cycling females were used for the localization and detection of the oviductin in oviductal oocytes and early embryos. Thin sections of hamster 1-cell oocytes, and 2-cell, and 8-cell embryos, embedded in Lowicryl and then incubated with the monoclonal antibody against the oviductin, revealed a homogenous distribution of antigenic sites in the matrix of the zona pellucida. While immunogold labeling was completely absent in the cytoplasm of 1-cell ova prior to fertilization, labeling was found associated with coated pits and coated vesicles and with flocculent material in the perivitelline space of fertilized eggs. In addition to these labeled endocytic structures, many endosomes, multivesicular bodies, and secondary lysosomes were also found to be labeled heavily in the cytoplasm of developing embryos. Our results indicate that following the transfer of Hamster Oviductin-1 to the zona pellucida of oocytes during transit in the oviduct and subsequent to fertilization, some of the oviductin associated with the zona pellucida appears to be internalized by blastomeres of the embryo and further processed through the endosomal/lysosomal pathway.


Erythroleukemia treated effects of rat plasma profile and erythrocyte membranes.
Erythroleukemia disease is caused by over production of malignant blood and immature large number of blood cells enters into peripheral compartment. Biophysical and biochemical changes in plasma and erythrocyte membrane in erythroleukemia treated rats were identified. Our study, leukemia is experimentally exposed in rats were injecting erythroleukemia cells (FLC) (H-2d) intravenously in adult rats and normal control rats were maintained. Significant increase in the activity of blood glucose, proteins levels, aspartate transaminase (AST) and alanine transaminase (ALT) values and significant decrease in haemoglobin (Hb), albumin levels in erythroleukemia treated rats were observed when compared with control rats. Cholesterol and low density liproprotein (LDL) levels increased significantly in erythroleukemia treated rats but triglycerides, high density lipoprotein (HDL) and very low density lipoprotein (VLDL) levels decreased significantly. Levels of red cell membrane cholesterol decreased in erythroleukemia treated rats in comparison with control while levels of phospholipids and proteins increased in erythrocytes of erythroleukemia treated rats. Red blood cell (RBC) and white blood cell (WBC) counts increased significantly and platelet count decreased. C/P (cholesterol/phospholipid) ratio decreased significantly in erythroleukemia treated rats. This study has been undertaken for the first time to investigate the effect of (FLC) (H-2d) erythroleukemia cells (treated) in intravenously in adult rats and normal control rats. Results indicate biophysical and biochemical alterations at molecular level in plasma and erythrocyte membrane.


How phytohormone IAA and chelator EDTA affect lead uptake by Zn/Cd hyperaccumulator Picris divaricata.
In this paper, the effects of indole-3-acetic acid (IAA) and/or ethylenediaminetetraacetic acid (EDTA) on lead uptake by a Zn/Cd hyperaccumulator Picris divaricata were studied. P. divaricata responded to Pb by better root system and increased biomass in presence of phytohormone IAA, which was able to reduce the inhibiting effects of Pb on transpiration without reducing the uptake of Pb The application of 100 microM IAA increased plant transpiration rate by about 20% and Pb concentration in leaves by about 37.3% as compared to treatment exposed to Pb alone. The enhanced phytoextraction efficiency could be attributed to the mechanisms played by IAA through alleviating Pb toxicity, creating better root system and plant biomass, promoting a higher transpiration rate as well as regulating the level of nutrient elements. On the contrary, inefficiency of phytoextraction was found with EDTA or the combination of IAA and EDTA probably because most Pb was in the form of Pb-EDTA complex which blocked the uptake by P. divaricata. The present study demonstrated that IAA was able to enhance the phytoextraction of Pb by Zn/Cd hyperaccumulator P. divaricata, providing a feasible method for the phytoremediation of polymetallic contaminated soils.


Standardized LC-MS/MS based steroid hormone profile-analysis.
In order to overcome many limitations of immunoassays, high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) has the potential to find its place in the clinical laboratory medicine for quantification of steroid hormones. A prerequisite for the application of a new analytical procedure in clinical diagnostics is standardization to minimize analytical intra- and interlaboratory variability and inaccuracy. We evaluate a newly standardized HPLC-MS/MS assay in kit-format, developed for routine determination of 16 steroid hormones in human serum samples. Fifteen metabolites can be measured quantitatively, which include aldosterone, androstenedione, androsterone, corticosterone, cortisol, cortisone, 11-deoxycortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), 17β-estradiol (E2), estrone (E1), etiocholanolone, 17α-hydroxyprogesterone (17OHP), progesterone, and testosterone. 11-Deoxycorticosterone is the only compound rated as semi-quantitative in this kit. The sample preparation is performed by solid phase extraction (SPE) on a 96-well plate. The standardized assay has been validated for human serum in terms of lower and upper limit of quantification (LLOQ 0.01-32 ng/mL, ULOQ 5-8000 ng/mL), linear correlation coefficient of calibration (R(2)>0.9966), intra- and inter-day precision (intra-day 1.1-8.8%, inter-day 5.2-14.8% and 8.2-18.6% for 11-deoxycorticosterone), accuracy (intra-day 88.3-115.5% and 109.3-128.2% for 11-deoxycorticosterone, inter-day 91.4-117.2% and 102.3-137.1% for 11-deoxycorticosterone), analytical total error (3.6-17.8%), proficiency test accuracy (85.4-113.4%), recovery (68-99%), and metabolite stability (freeze/thaw stability 95.5-108.1%, short term stability 86.9-107.2%). Inter-assay comparison with a routine reference HPLC-MS/MS assay and seven immunoassays demonstrates the outstanding high performance of this HPLC-MS/MS based kit by improvements in accuracy for progesterone, androstenedione, and 17OHP. Finally, results of two metyrapone tests demonstrate the potential of the standardized HPLC-MS/MS assay for the analysis of a comprehensive steroid hormone profile in clinical diagnostics.


Excess Heritability Contribution of Alcohol Consumption Variants in the "Missing Heritability" of Type 2 Diabetes Mellitus.
We aim to compare the relative heritability contributed by variants of behavior-related environmental phenotypes and elucidate the role of these factors in the conundrum of "missing heritability" of type 2 diabetes. Methods: We used Linkage-Disequilibrium Adjusted Kinships (LDAK) and LDAK-Thin models to calculate the relative heritability of each variant and compare the relative heritability for each phenotype. Biological analysis was carried out for the phenotype whose variants made a significant contribution. Potential hub genes were prioritized based on topological parameters of the protein-protein interaction network. We included 16 behavior-related phenotypes and 2607 valid variants. In the LDAK model, we found the variants of alcohol consumption and caffeine intake were identified as contributing higher relative heritability than that of the random variants. Compared with the relative expected heritability contributed by the variants associated with type 2 diabetes, the relative expected heritability contributed by the variants associated with these two phenotypes was higher. In the LDAK-Thin model, the relative heritability of variants of 11 phenotypes was statistically higher than random variants. Biological function analysis showed the same distributions among type 2 diabetes and alcohol consumption. We eventually screened out 31 hub genes interacting intensively, four of which were validated and showed the upregulated expression pattern in blood samples seen in type 2 diabetes cases. Conclusion: We found that alcohol consumption contributed higher relative heritability. Hub genes may influence the onset of type 2 diabetes by a mediating effect or a pleiotropic effect. Our results provide new insight to reveal the role of behavior-related factors in the conundrum of "missing heritability" of type 2 diabetes.


Numerical generation of a polarization singularity array with modulated amplitude and phase.
A point having no defined polarized ellipse azimuthal angle (circularly polarized) in a space-variant vector field is called a polarization singularity, and it has three types: Lemon, Monstar, and Star. Recently, the connection of polarization singularities has been performed. Inspired by this, we conduct a numerical generation of a polarization singularity array. Our method is based on two orthogonal linearly polarized light beams with modulated amplitude and phase. With appropriate distribution functions of amplitudes and phases we can control the polarized states of polarization singularities, which offer a possibility to simulate a polarization singularity array.


Memory for faces, shoes, and objects by deaf and hearing signers and hearing nonsigners.
The visual spatial memory of 15 deaf signers, 15 hearing signers, and 15 hearing nonsigners for shoes, faces, and verbalizable objects was measured using the game Concentration. It was hypothesized that the deaf and hearing signers would require fewer attempts than the hearing nonsigners on the shoes and faces tasks because of their experience of using a visual-spatial language; and, in the case of the Deaf, also possibly, due to a compensatory mechanism. It was also hypothesized that memory for shoes would be more like that for faces than for simple objects. It was also anticipated that there would be no difference between the three group's memories for verbalizable objects. Deaf signers were found to be similar to hearing signers, both of whom were better than hearing nonsigners on the faces and shoes tasks. Generally, performance on the faces and shoes tasks was similar and followed the same pattern for the three groups. The three groups performed at a similar level on the objects task. There were no gender differences.


[Effect of a dialysis solution with icodextrin on ultrafiltration and selected metabolic parameters in patients treated with peritoneal dialysis].
To date, peritoneal dialysis has been performed almost exclusively using dialysis solutions containing glucose as the osmotic agent. Use of these solutions is fraught with problems regarding adequate fluid removal from the body and is also associated with undesirable metabolic effects; hence the search for alternative osmotic agents. A dialysis solution with the glucose polymer icodextrin generates ultrafiltration on the principle of colloidal osmosis. The aim of the study was to establish the effect of icodextrin-base dialysis solution on the magnitude of ultrafiltration and evaluate selected metabolic parameters of patients treated by ambulatory peritoneal dialysis. A total of 9 patients whose glucose-based solution was replaced by an icodextrin-based solution during the night-time exchange were evaluated. A control group of 9 patients used glucose-solution during all exchanges. Night-time bag ultrafiltration, blood pressure, and the serum levels of lipids, insulin, leptin, maltose, and amylase were determined before icodextrin administration (time 0), at one-month intervals (time 1, 2, 3), and one month after study completion (time 4). In icodextrin-treated patients, ultrafiltration rose from 246.5 +/- 60.5 ml (mean +/- SEM) at time 0 to 593.1 +/- 87.4 ml; p < 0.01, at time 1, to 547 +/- 67 ml; p < 0.05, at time 2, and to 586.7 +/- 58.8 ml; p < 0.01, at time 3, the icodextrin administration led to a rise in maltose from 0.02 +/- 0.01 g/l at time 0 to 0.1 +/- 0.1 g/l; p < 0.01, at time 1, to 1.0 +/- 0.09 g/l; p < 0.01, at time 2, and to 1.1 +/- 0.09 g/l; p < 0.01, at time 3, with a fall to zero values at time 4 (NS). Icodextrin administration was followed by a decrease in leptinemia from 34.6 +/- 17.2 ng/ml at time 0 to 21.7 +/- 8.9 ng/ml; p < 0.05, at time 1, to 21.4 +/- 9.5 ng/ml; p < 0.05, at time 2, and to 15.9 +/- 24.1 ng/ml; p < 0.05 at time 4. Insulin and lipid levels were not affected. There was no change in the above parameters in the control group. Icodextrin-treated patients reduced their antihypertensive medication, but not statistically significantly. Icodextrin administration significantly increase ultrafiltration thus providing for effective control of hydration status without the need for high-level glucose-based dialysis solutions. The use of a glucose polymer-based dialysis solution is associated with a significant yet reversible rise in serum maltose. The decrease in leptin may signal a reduction in body weight after replacing glucose in dialysis solutions with icodextrin, or enhanced rates of leptin elimination as a result of ultrafiltration-induced convective transport.


Modelling of recharge and pollutant fluxes to urban groundwaters.
Urban groundwater resources are of considerable importance to the long-term viability of many cities world-wide, yet prediction of the quantity and quality of recharge is only rarely attempted at anything other than a very basic level. This paper describes the development of UGIf, a simple model written within a GIS, designed to provide estimates of spatially distributed recharge and recharge water quality in unconfined but covered aquifers. The following processes (with their calculation method indicated) are included: runoff and interception (curve number method); evapotranspiration (Penman-Grindley); interflow (empirical index approach); volatilization (Henry's law); sorption (distribution coefficient); and degradation (first order decay). The input data required are: meteorological data, landuse/cover map with event mean concentration attributes, geological maps with hydraulic and geochemical attributes, and topographic and water table elevation data in grid form. Standard outputs include distributions of: surface runoff, infiltration, potential recharge, ground level slope, interflow, actual recharge, pollutant fluxes in surface runoff, travel times of each pollutant through the unsaturated zone, and the pollutant fluxes and concentrations at the water table. The process of validation has commenced with a study of the Triassic Sandstone aquifer underlying Birmingham, UK. UGIf predicts a similar average recharge rate for the aquifer as previous groundwater flow modelling studies, but with significantly more spatial detail: in particular the results indicate that recharge through paved areas may be more important than previously thought. The results also highlight the need for more knowledge/data on the following: runoff estimation; interflow (including the effects of lateral flow and channelling on flow times and therefore chemistry); evapotranspiration in paved areas; the nature of unsaturated zone flow below paved areas; and the role of the pipe network. Although considerably more verification is needed, UGIf shows promise for use: in providing input for regional groundwater solute transport models; in identifying gaps in knowledge and data; in determining which processes are the most important influences on urban groundwater quantity and quality; in evaluating existing recharge models; in planning, for example in investigation of the effects of landuse or climate change; and in assessing groundwater vulnerability.


Relation between functions and conformational characteristics of modified nucleosides found in tRNAs.
Conformational characteristics of N4-acetyl-2'-O-methylcytidine (ac4Cm), 5-methyl-2'-O-methylcytidine (m5Cm) and N2-dimethyl-2'-O-methylguanosine (m2(2)Gm) found in tRNAs from extremely thermophilic archaebacteria were analyzed by proton NMR spectroscopy. The 2'-O-methylation of ac4C, m5C and m2(2)G was found to stabilize the C3'-endo form and therefore cause "conformational rigidity". In particular, the ac4Cm was found to be extremely rigid due to additive effects of the N4-acetylation and 2'-O-methylation. Therefore, tRNAs from the extremely thermophilic archaebacteria use the base modifications in combination with the 2'-O-methylation, resulting in stabilization of the A-type conformation at specific positions in the tRNAs even at very high temperatures. In contrast, mesophile tRNAs use for a given site only one of these ribose and base modifications each of which is effective enough by itself at ordinary temperatures. These findings are consistent with our previous findings that roles of a variety of post-transcriptional modifications are to regulate the conformational rigidity/flexibility which is essential for the tRNA functions.


How much does climate change threaten European forest tree species distributions?
Although numerous species distribution models have been developed, most were based on insufficient distribution data or used older climate change scenarios. We aimed to quantify changes in projected ranges and threat level by the years 2061-2080, for 12 European forest tree species under three climate change scenarios. We combined tree distribution data from the Global Biodiversity Information Facility, EUFORGEN, and forest inventories, and we developed species distribution models using MaxEnt and 19 bioclimatic variables. Models were developed for three climate change scenarios-optimistic (RCP2.6), moderate (RCP4.5), and pessimistic (RPC8.5)-using three General Circulation Models, for the period 2061-2080. Our study revealed different responses of tree species to projected climate change. The species may be divided into three groups: "winners"-mostly late-successional species: Abies alba, Fagus sylvatica, Fraxinus excelsior, Quercus robur, and Quercus petraea; "losers"-mostly pioneer species: Betula pendula, Larix decidua, Picea abies, and Pinus sylvestris; and alien species-Pseudotsuga menziesii, Quercus rubra, and Robinia pseudoacacia, which may be also considered as "winners." Assuming limited migration, most of the species studied would face a significant decrease in suitable habitat area. The threat level was highest for species that currently have the northernmost distribution centers. Ecological consequences of the projected range contractions would be serious for both forest management and nature conservation.


Analysis of phenolic compounds in health care products by low-pressure liquid-chromatography with monolithic column and chemiluminescent detection.
This paper presents a new application for monolithic columns with low-pressure chromatographic separation using an flow injection analysis configuration with chemiluminescent detection for the determination of a mixture of phenolic compounds: phloroglucinol, 2,4-dihydroxybenzoic acid, salicylic acid, methyl paraben and n-propyl gallate. The procedure consists of the separation of these compounds on a reverse-phase ultra-short monolithic column with pH 3.0 acetate buffer and 5% acetonitrile as carrier phase. The detection is based on a chemiluminescence measurement coming from Ce(IV)-Rhodamine 6G chemistry with the incorporation of two different chemiluminescent chemical conditions in the chromatographic setup in order to enhance the sensitivity for the different phenolic compounds. All separation and detection variables were optimized to propose a determination method. The analysis is performed in 280?s, with the sampling frequency being some 13 h(-1) . The calibration function is a double reciprocal function obtaining good results within two orders of magnitude. The limits of detection were 8.8 × 10 (-8) m (phloroglucinol), 2.7 × 10 (-8) m (2,4-dihydroxybenzoic acid); 2.3 × 10 (-8) m (salicylic acid); 5.2 × 10 (-8) m (methyl paraben) and 4.1 × 10 (-6) m (n-propyl gallate), and the relative standard deviations at a medium level of the linear range were 4.4% (phloroglucinol), 2.8% (2,4-dihydroxybenzoic acid), 5.2% (salicylic acid), 3.6% (methyl paraben) and 6.8% (n-propyl gallate). The method was applied and validated satisfactorily for the determination of these compounds in healthcare products, comparing the results against an HPLC reference method.


Gastrointestinal ischemia related mortality in patients undergoing off- or on-pump coronary artery bypass grafting.
Gastrointestinal ischemia (GII) after heart surgery is a rare but devastating condition. The aim of this study was to compare the occurrence of GII after coronary artery bypass grafting (CABG) performed off-pump (OPCAB) vs on-pump (ONCAB). We retrospectively evaluated 2625 adult patients who underwent isolated coronary artery surgery during a 6-year period. The OPCAB group included 658 patients and the ONCAB group 1967 patients. Patients were evaluated, and GII, morbidity, and mortality in the 2 groups were compared. GII developed in 0.4% (7 of 1967) patients in the ONCAB group and in 0.2% (1 of 658) patients in the OPCAB group (P = .28). Mortality rates due to GII were 0.2% (4 of 1967) in the ONCAB group, and no deaths occurred in the OPCAB group (P < .04). Postoperative atrial fibrillation incidence with GII was 100% (7 of 7) in ONCAB group and 0% (0 of 1) in the OPCAB group (P < .01). Compared to ONCAB, the OPCAB procedure has lower GII related mortality rates, which is an important cause of morbidity and mortality in the postoperative period of CABG surgery.


Involvement of Mammalian RF-Amide Peptides and Their Receptors in the Modulation of Nociception in Rodents.
Mammalian RF-amide peptides, which all share a conserved carboxyl-terminal Arg-Phe-NH2 sequence, constitute a family of five groups of neuropeptides that are encoded by five different genes. They act through five G-protein-coupled receptors and each group of peptide binds to and activates mostly one receptor: RF-amide related peptide group binds to NPFFR1, neuropeptide FF group to NPFFR2, pyroglutamylated RF-amide peptide group to QRFPR, prolactin-releasing peptide group to prolactin-releasing peptide receptor, and kisspeptin group to Kiss1R. These peptides and their receptors have been involved in the modulation of several functions including reproduction, feeding, and cardiovascular regulation. Data from the literature now provide emerging evidence that all RF-amide peptides and their receptors are also involved in the modulation of nociception. This review will present the current knowledge on the involvement in rodents of the different mammalian RF-amide peptides and their receptors in the modulation of nociception in basal and chronic pain conditions as well as their modulatory effects on the analgesic effects of opiates.


Abdominal imaging can misdiagnose submassive hepatic necrosis as cirrhosis in acute liver failure.
Patients with acute liver failure (ALF) can be listed status I for liver transplantation (LT) whereas patients with cirrhosis must follow the MELD scoring system. Liver imaging can mistakenly diagnose submassive hepatic necrosis in ALF as cirrhosis. The purpose of our study was to assess the accuracy of ultrasound (US) and computed tomography (CT) in distinguishing cirrhosis from ALF. All patients listed for ALF and transplanted during the study period were included. Controls were age- and gender-matched cirrhotic patients who underwent LT during the same period. Abdominal US or CT scans obtained on all patients were independently reviewed by three blinded abdominal radiologists. Explants from all patients were reviewed by two blinded pathologists, and histological diagnosis was correlated with radiological diagnosis. Forty-one patients with ALF and 42 patients with cirrhosis were analyzed. Univariate and multivariate analyses both revealed overall accuracy of 85% for ultrasound and 93% for CT. US and CT scans both provide high levels of accuracy in terms of discriminating ALF from cirrhosis but measures taken to determine whether a patient has ALF vs. cirrhosis needs to approach 100% accuracy. Thus, imaging studies alone should not definitively diagnosis one etiology of liver failure over the other.


Effects of disodium dichloromethylene diphosphonate on bone loss in paraplegic patients.
21 paraplegic patients with recent traumatic spinal cord injury were orally administered 400 (n = 7) or 1,600 (n = 7) mg/d of disodium dichloromethylene diphosphonate (Cl2MDP) and compared with a placebo group (n = 7) to test the preventive effects of the drug on acute bone loss and osteoclastic resorption. Cl2MDP therapy was initiated at a mean of 17.6 d after the onset of paraplegia. The study lasted at least 6 mo, consisting of a 3.5-mo treatment period, and a variable follow-up period. The effects of Cl2MDP were assessed by blood and urine biochemistry, bone histomorphometry on transilial samples, photon absorptiometry of the tibia and fibula, and radiomorphometry of the femur. The elevation in serum and urinary calcium and in urine hydroxyproline observed in the placebo group did not appear under treatment. With both doses of Cl2MDP there was no further decrease in the bone mineral content. In the treated groups, a smaller percentage increase in osteoclastic population was also noted when compared with the placebo group, but this difference was not significant. There was no mineralization defect induced by Cl2MDP, as shown by tetracycline double labeling. It thus appears that at doses ranging between 400 and 1,600 mg, given as early as possible, Cl2MDP can prevent or reduce the development of the acute bone loss of paraplegic patients, without adverse side effects, though it does not prevent the development of heterotopic ossification.


Intrauterine ischemic reperfusion switches the fetal transcriptional pattern from HIF-1α- to P53-dependent regulation in the murine brain.
Ischemic reperfusion (IR) during the perinatal period is a known causative factor of fetal brain damage. So far, both morphologic and histologic evidence has shown that fetal brain damage can be observed only several hours to days after an IR insult has occurred. Therefore, to prevent fetal brain damage under these circumstances, a more detailed understanding of the underlying molecular mechanisms involved during an acute response to IR is necessary. In the present work, pregnant mice were exposed to IR on day 18 of gestation by clipping one side of the maternal uterine horn. Simultaneous fetal electrocardiography was performed during the procedure to verify that conditions resulting in fetal brain damage were met. Fetal brain sampling within 30 minutes after IR insult revealed molecular evidence that a fetal response was indeed triggered in the form of inhibition of the Akt-mTOR-S6 synthesis pathway. Interestingly, significant changes in mRNA levels for both HIF-1α and p53 were apparent and gene regulation patterns were observed to switch from a HIF-1α-dependent to a p53-dependent process. Moreover, pre-treatment with pifithrin-α, a p53 inhibitor, inhibited protein synthesis almost completely, revealing the possibility of preventing fetal brain damage by prophylactic pifithrin-α treatment.


Engineering Native and Synthetic Pathways in Pseudomonas putida for the Production of Tailored Polyhydroxyalkanoates.
Growing environmental concern sparks renewed interest in the sustainable production of (bio)materials that can replace oil-derived goods. Polyhydroxyalkanoates (PHAs) are isotactic polymers that play a critical role in the central metabolism of producer bacteria, as they act as dynamic reservoirs of carbon and reducing equivalents. PHAs continue to attract industrial attention as a starting point toward renewable, biodegradable, biocompatible, and versatile thermoplastic and elastomeric materials. Pseudomonas species have been known for long as efficient biopolymer producers, especially for medium-chain-length PHAs. The surge of synthetic biology and metabolic engineering approaches in recent years offers the possibility of exploiting the untapped potential of Pseudomonas cell factories for the production of tailored PHAs. In this article, an overview of the metabolic and regulatory circuits that rule PHA accumulation in Pseudomonas putida is provided, and approaches leading to the biosynthesis of novel polymers (e.g., PHAs including nonbiological chemical elements in their structures) are discussed. The potential of novel PHAs to disrupt existing and future market segments is closer to realization than ever before. The review is concluded by pinpointing challenges that currently hinder the wide adoption of bio-based PHAs, and strategies toward programmable polymer biosynthesis from alternative substrates in engineered P. putida strains are proposed.


Prefrontal Lobe and Posterior Cingulate Cortex Activations in Patients with Major Depressive Disorder by Using Standardized Weighted Low-Resolution Electromagnetic Tomography.
The differences in brain activity between patients with major depressive disorder (MDD) and healthy adults have been confirmed by functional magnetic resonance imaging (fMRI), positron emission tomography (PET), and electroencephalography (EEG). The prefrontal lobe and posterior cingulate cortex (PCC) are related to emotional regulation in patients with MDD. However, the high cost and poor time resolution of fMRI and PET limit their clinical application. Recently, researchers have used high time resolution of standardized weighted low-resolution electromagnetic tomography (swLORETA) to investigate deep brain activity. This study aimed to convert raw EEG signals into swLORETA images and explore deep brain activity in patients with MDD and healthy adults. BrainMaster EEG equipment with a 19-channel EEG cap was used to collect resting EEG data with eyes closed for 5 min. NeuroGuide software was used to remove the EEG artifacts, and the swLORETA software was used to analyze 12,700 voxels of current source density (CSD) for 139 patients with MDD and co-morbid anxiety symptoms (mean age = 43.08, SD = 13.76; 28.78% were male) and 134 healthy adults (mean age = 40.60, SD = 13.52; 34.33% were male). Deep brain activity in the frontal lobe and PCC at different frequency bands was analyzed, including delta (1-4 Hz), theta (5-7 Hz), alpha (8-11 Hz), beta (12-24 Hz), beta1 (12-14 Hz), beta2 (15-17 Hz), beta3 (18-24 Hz), and high beta (25-29 Hz). There was lower delta and theta and higher beta, beta1, beta2, beta3, and high-beta activity at the prefrontal lobe (dorsal medial prefrontal cortex [dmPFC], ventral medial prefrontal cortex [vmPFC], and dorsal lateral prefrontal cortex [dlPFC], ventral lateral prefrontal cortex [vlPFC], orbital frontal cortex [OFC]) and PCC in MDD patients compared with healthy adults. There was no significant difference in alpha activity between the two groups. This study indicates brain hyperactivity in the right prefrontal lobe (dlPFC and vmPFC) and PCC in patients with MDD with co-morbid anxiety symptoms, and the dlPFC and PCC were also related to emotion regulation in MDD. Inhibiting high-beta activity or restoring delta and theta activity to the normal range in the right frontal lobe and PCC may be possible in z-score neurofeedback protocols for patients with MDD in future studies.


Photographic assessment of head shape following sagittal synostosis surgery.
A photographic assessment of the head shape of infants who had undergone surgical correction of sagittal synostosis was performed to determine (a) whether this subset could be delineated from an age-matched normal subpopulation and (b) whether two operative procedures differed in achieving normalization of head shape. This retrospective study included 8 patients who underwent extended strip craniectomy, 12 patients who underwent subtotal calvarectomy and cranial vault remodeling, and 12 age-matched subjects with no calvarial abnormality, for a total of 32 subjects. Criteria for inclusion in this study included surgery for sagittal synostosis within the first year of life and postoperative photographs at ages 4 to 8 years (mean, 4.5 years). Each set of images (frontal and lateral profile) were ranked from most to least normal by five lay observers and four professional observers. The rankings were analyzed with statistics designed for ordinal data. Differences in ranking between treatment groups were examined with Kruskal-Wallis rank sums tests. Mean ranks were calculated for lay and professional observers in an attempt to produce simpler and more generalizable results; these means were also analyzed using statistics designed for ordinal data. There was no statistical difference in the ranks of infants who had undergone a surgical correction and the normal subpopulation. In the mean rankings of the lay observers, the normal groups had the highest score mean (15.6), the group with extended strip craniectomy was second (16.0), and the subtotal calvarectomy with calvarial remodeling group was last (17.8) (p = 0.84). In the mean rankings of the professional observers, the normal groups again had the highest score mean (15.8), the subtotal calvarectomy group was second (15.9), and the extended craniectomy group was last (18.6) (p = 0.77). These results suggest that children who have undergone correction of sagittal synostosis in infancy are indistinguishable from their peers, on the basis of fully haired head shape on frontal and lateral photographs, when they begin primary school, irrespective of the type of calvarial surgery.


Metabolic profiles and genetic diversity of denitrifying communities in activated sludge after addition of methanol or ethanol.
External carbon sources can enhance denitrification rates and thus improve nitrogen removal in wastewater treatment plants. The effects of adding methanol and ethanol on the genetic and metabolic diversity of denitrifying communities in activated sludge were compared using a pilot-scale plant with two parallel lines. A full-scale plant receiving the same municipal wastewater, but without external carbon source addition, was the reference. Metabolic profiles obtained from potential denitrification rates with 10 electron donors showed that the denitrifying communities altered their preferences for certain compounds after supplementation with methanol or ethanol and that methanol had the greater impact. Clone libraries of nirK and nirS genes, encoding the two different nitrite reductases in denitrifiers, revealed that methanol also increased the diversity of denitrifiers of the nirS type, which indicates that denitrifiers favored by methanol were on the rise in the community. This suggests that there might be a niche differentiation between nirS and nirK genotypes during activated sludge processes. The composition of nirS genotypes also varied greatly among all samples, whereas the nirK communities were more stable. The latter was confirmed by denaturing gradient gel electrophoresis of nirK communities on all sampling occasions. Our results support earlier hypotheses that the compositions of denitrifier communities change during predenitrification processes when external carbon sources are added, although no severe effect could be observed from an operational point of view.


Simulation as a Tool for Teaching and Learning Epidemiologic Methods.
In aspiring to be discerning epidemiologists, we must learn to think critically about the fundamental concepts in our field and be able to understand and apply many of the novel methods being developed today. We must also find effective ways to teach both basic and advanced topics in epidemiology to graduate students, in a manner that goes beyond simple provision of knowledge. Here, we argue that simulation is one critical tool that can be used to help meet these goals, by providing examples of how simulation can be used to address 2 common misconceptions in epidemiology. First, we show how simulation can be used to explore nondifferential exposure misclassification. Second, we show how an instructor could use simulation to provide greater clarity on the correct definition of the P value. Through these 2 examples, we highlight how simulation can be used to both clearly and concretely demonstrate theoretical concepts, as well as to test and experiment with ideas, theories, and methods in a controlled environment. Simulation is therefore useful not only in the classroom but also as a skill for independent self-learning.


Altretamine.
Altretamine is a National Cancer Institute-designated group C antineoplastic agent used in the treatment of advanced ovarian cancer. Altretamine is a highly lipid-soluble drug available only for oral administration as a capsule. The drug is activated through metabolic oxidation to intermediate methylol derivatives and formaldehyde. It is unclear which metabolite is the major species responsible for cytotoxicity or the primary mechanism of cytotoxicity. As a single agent in the treatment of ovarian cancer, altretamine demonstrates a response rate similar to other active agents in this disease (21-39 percent). The major utility of altretamine is in combination with other agents such as cyclophosphamide, doxorubicin, fluorouracil, melphalan, and cisplatin. However, few randomized trials have evaluated the contribution of altretamine in these multiagent combinations. Dose-limiting toxicities include gastrointestinal (nausea, vomiting, anorexia), hematologic, and neurotoxic (peripheral neurotoxicity). The therapeutic role of altretamine is limited because of a toxicity profile similar to that of cisplatin, one of the more active agents in ovarian cancer. Its use should be reserved for patients who are not candidates for more standard platinum-based regimens.


Cognitive changes in prevalent and incident cardiovascular disease: a 12-year follow-up in the Maastricht Aging Study (MAAS).
Cardiovascular disease (CVD) has been suggested to accelerate cognitive decline and to be a risk factor for dementia, but still little is known about the cognitive course after a first cardiovascular event. Therefore, the present study aims to investigate the cognitive trajectories in both prevalent and incident CVD over a 12-year time period in the general population. Cognitively healthy participants (age 24-82 years, n = 1823) of a prospective cohort study were serially assessed at baseline, 6 and 12 years. Verbal memory, executive function, and information processing speed were analysed in adults with prevalent, incident, and no CVD. Random effects models were used to test the association between CVD and change in cognitive function over time. At baseline, participants with prevalent CVD showed more decline in memory and information processing speed than healthy controls. Participants with incident CVD also showed more decline in these cognitive domains, but this was only significant in the follow-up period from 6 to 12 years. Associations were more pronounced in participants aged younger than 65 years at baseline, and in sub-analyses with angina pectoris or myocardial infarction as the most prevalent CVD conditions. Prevalent and incident CVD predict cognitive decline in middle-aged individuals. Findings for incident CVD suggest that the onset of decline is linked in time with the vascular event itself. Timely CVD management may delay the onset of decline.


Training of patients for self-management of oral anticoagulant therapy: standards, patient suitability, and clinical aspects.
Self-control and self-management of oral anticoagulant therapy have become more and more attractive for patients undergoing long-term treatment. In our training center, we examined 50 patients who took part in a standardized training course for self-management. Patients (36 men, 14 women) were preselected according to the guidelines of the German Association for Self-management of Oral Anticoagulation (ASA e.V.) and were all trained by the same physician. The complete course took an average of eight sessions. Patients older than 59 years needed significantly more training time in theoretical advising than younger patients; they did not need more training time in practical matters. The values between International Normalized Ratio (INR) measured in venous blood samples and by self-assessment were comparable for both groups. There was a good overall correlation between self-controlled INRs and laboratory assays, however, the self-assayed INRs were significantly lower than those from the venous blood samples.


Ex vivo expansion of natural killer cells using cryopreserved irradiated feeder cells.
Currently, feeder cells are γ-irradiated immediately before use for the ex vivo expansion of natural killer (NK) cells from human peripheral blood. Storing irradiated feeder cells by cryopreserving them in multiple vials would be more convenient than irradiating cells each time they are needed. We compared NK cell expansion using cryopreserved-irradiated feeder cells (cryopreserved group) and freshly-irradiated feeder cells (fresh group). To expand NK cells, peripheral blood mononuclear cells were isolated and co-cultured with-100 Gy-irradiated K562 leukemia cells that had been modified to express 4-1BB ligand and membrane-bound (mb) interleukin (IL)-15 (K562-mb15-41BBL cells) for three weeks in the presence of IL-2 and IL-15. Fresh and cryopreserved K562-mb15-41BBL feeder cells expressed similar levels of 4-1BB ligand, whereas membrane-bound IL-15 expression was lower in the cryopreserved cells than in the fresh cells. The NK cell expansion rate did not differ between the two groups (980-fold vs. 1058-fold, respectively), although the mean NK cell purity was higher in the fresh-group than in the cryopreserved-group at day 14 (94.1% vs. 92.5%, respectively) and day 21 (97.1% vs. 95.4%, respectively). The NK cells from the two feeder cell groups did not differ in cytotoxicity against various malignant cell lines at effector-to-target ratios of 4:1, 2:1, and 1:1, or in the expression pattern of NK cell receptors [cluster of differentiation (CD)-16, natural killer group-2, member D (NKG2D), CD69, NKp30, NKp44, NKp46, and CD158b] and level of interferon-γ secretion. Our results demonstrate that cryopreserved irradiated feeder cells can be used for the ex vivo expansion of human NK cells and provide a convenient improvement on current methods.


Effect of Microstructures of Carbon Nanoproducts Grown on Carbon Fibers on the Interfacial Properties of Epoxy Composites.
Two kinds of carbon nanoproducts with different microstructures, namely, carbon nanotubes (CNTs) and carbon nanofibers (CNFs), were grown on the surface of carbon fibers (CFs) by chemical vapor deposition (CVD) at low temperatures to improve the interface bonding between fibers and resins. The short-beam method and the micro-debonding method were used to test the interlaminar shear strength (ILSS) and interfacial shear strength (IFSS) of the composites. The results showed that the contribution of CNTs to the improvement of interfacial properties was better than that of CNFs. Specifically, the ILSS and IFSS of the CF-CNFs/epoxy composites increased by 18.59 and 24.39%, respectively, while the ILSS and IFSS of the CF-CNTs/epoxy composites increased by 26.97 and 47.79%, respectively. Compared with CNFs, the high degree of graphitization of CNTs and the π-interactions with the resin can better induce the formation of an interphase between the fiber and the resin, which suppressed the initiation of cracks and extended the propagation path of the cracks in the composites.


Traumatic abdominal wall hernia.
Traumatic abdominal hernias are uncommon. A case is presented of a 49-year-old man who struck his abdomen on the handlebars of a snowmobile. Local pain, bruising, and reducible swelling with cough impulse suggest the diagnosis. Lateral or oblique X-rays and use of contrast material are recommended. Differentiation from a preexisting hernia may be important medicolegally. Incision, if necessary, should be very carefully made. In the present case surgical repair was followed by uneventful recovery.


Effects of cyclic nucleotides on the longitudinal internal resistance in the rabbit sinus node.
The effect of dibutyryl cyclic AMP as well as dibutyryl cyclic GMP on the longitudinal internal resistance was studied in the rabbit sinus node strips by means of microelectrode and sucrose gap techniques. Dibutyryl cyclic AMP added to the sucrose gap decreased the resistance and increased the spontaneous rate whereas dibutyryl cyclic GMP evoked the opposite effects. These results suggest that cyclic nucleotides influence the cell coupling in the sinus node.


Modern multiphase catalysis: new developments in the separation of homogeneous catalysts.
Homogeneous catalysts are powerful tools for the synthesis of fine chemicals, pharmaceuticals and materials, however their exploitation on an industrial scale is often held back due to the challenges of separating and recycling the catalyst. This perspective focuses on approaches to multiphase catalysis that have emerged in the last decade, highlighting methods that can address the separation issues and in some cases result in superior catalyst performance and environmental benefits.


Clinical Outcomes of Donor Sites After Osteochondral Graft Harvest From Healthy Knees.
The use of autologous osteochondral grafts has become popular in the treatment of small, isolated, well-contained articular cartilage defects. However, donor site morbidity is a problem, and few reports are available of donor site morbidity after mosaicplasty. To examine the clinical outcomes of donor sites after osteochondral grafts from healthy knees. Case series; Level of evidence, 4. Between September 1997 and September 2011, there were 40 patients (40 knees; 32 men, 8 women; 31 right knees, 9 left knees) with asymptomatic osteochondral graft donor sites used for autologous transfer; all had a follow-up period of >2 years. The mean patient age at surgery was 21.0 years (range, 12-58 years). The recipient sites included the elbow (n = 28), contralateral knee (n = 5), and ankle (n = 7). The mean diameter of the grafted plugs was 7.5 mm (range, 4.5-9 mm), and the mean number of grafted plugs was 2.2 (range, 1-3). At a mean follow-up of 43.1 months (range, 24-177 months), knee symptoms, return to sport, ability to sit straight in Japanese style, and radiological changes of the patellofemoral joint were evaluated. Whether operative age, follow-up period, and diameter or number of the grafted plugs were risk factors was analyzed statistically. Significance was defined as P < .05. Thirty-four patients had no knee symptoms, and 4 patients had occasional mild knee pain. Two patients underwent reoperation for arthrofibrosis and not for cartilage defect. Twenty-seven patients had complete return to sports, and 32 patients could sit straight; donor site morbidity was not the cause of failure to return to sports or inability to sit straight. The radiological changes became worse in 3 patients, and the risk factor for degenerative change was older operative age. When osteochondral plugs were obtained from healthy knees, 34 patients (85%) were asymptomatic at follow-up. No donor site defects required surgical intervention due to persistent symptoms.


A new model of myofibroblast-cardiomyocyte interactions and their differences across species.
Although coupling between cardiomyocytes and myofibroblasts is well known to affect the physiology and pathophysiology of cardiac tissues across species, relating these observations to humans is challenging because the effect of this coupling varies across species and because the sources of these effects are not known. To identify the sources of cross-species variation, we built upon previous mathematical models of myofibroblast electrophysiology and developed a mechanoelectrical model of cardiomyocyte-myofibroblast interactions as mediated by electrotonic coupling and transforming growth factor-β1. The model, as verified by experimental data from the literature, predicted that both electrotonic coupling and transforming growth factor-β1 interaction between myocytes and myofibroblast prolonged action potential in rat myocytes but shortened action potential in human myocytes. This variance could be explained by differences in the transient outward K+ current associated with differential Kv4.2 gene expression across species. Results are useful for efforts to extrapolate the results of animal models to the predicted effects in humans and point to potential therapeutic targets for fibrotic cardiomyopathy.


Differential interactions of urocortin/corticotropin-releasing hormone peptides with the blood-brain barrier.
The two newest members of the urocortin (UCN)/corticotropin-releasing hormone (CRH) family of peptides - UCN II and UCN III - bind to the CRH-2 receptor, suppress feeding, and are expressed in the periphery as well as the brain. We used several sensitive techniques to examine their interactions with the blood-brain barrier (BBB). Of the four known peptides in this family, each interacts with the BBB differently. UCN I barely enters the brain from blood unless its latent saturable influx system is activated by leptin or pretreatment with glucose. However, neither leptin nor glucose affected the entry of intact UCN II. UCN II reached brain paranchyma at a moderate rate that was not self-inhibited or cross-inhibited by UCN/CRH peptides. The apparent, but misleading, rapid influx of UCN III (stresscopin) could be explained by degradation at the BBB itself. Influx of CRH into brain was slower than UCN II but faster than UCN I; it was inhibited by excess CRH but not by excess UCN I, II, III, or leptin. CRH is the only member of this family to have a saturable efflux system out of the brain. Determination of hydrogen bonding, newly applied here to ingestive peptides, was not helpful in explaining these differential interactions of the UCN peptides with the BBB.


Clinical characteristics and prognosis of patients with Pneumocystis jirovecii pneumonia without a compromised illness.
Pneumocystis jirovecii pneumonia (PCP) is a fatal respiratory infection, mostly associated with immunocompromised conditions. Several reports have described PCP development in patients who were not immunocompromised, but the clinical course and prognosis of PCP are not well understood. We compared the clinical characteristics and prognoses between patients with and without immunocompromised conditions who developed PCP. We retrospectively analyzed patients who had been treated for PCP from three hospitals. We defined immunocompromised (IC) status as following: human immunodeficiency virus (HIV) infection; hematological malignancy; solid organ tumor under chemotherapy; rheumatic disease; medication with immunosuppressive agents. Patients without immunocompromised status were defined as being non-immunocompromised (non-IC). The IC and non-IC groups comprised 173 and 14 patients. The median ages were 62.0 and 74.0 years in the IC and the non-IC group, respectively. The median interval between admission and anti-PCP treatment was significantly longer for patients in the non-IC group than that for patients in the IC group (7 vs. 2 days). The in-hospital mortality rates were significantly higher for patients in the non-IC group than that for patients in the IC group (71.4% vs. 43.9%; P = 0.047). A longer interval between admission and anti-PCP therapy was associated with increased 90-day mortality rate in patients with PCP (hazard ratio, 1.082; 95% confidence interval, 1.015-1.153; P = 0.016). Patients with PCP with no predisposing illnesses were older and had higher mortality rates than IC patients with PCP. Delayed anti-PCP treatment was associated with increased 90-day mortality.


Functional characterization of a GJA1 frameshift mutation causing oculodentodigital dysplasia and palmoplantar keratoderma.
A frameshift mutation generated from a dinucleotide deletion (780-781del) in the GJA1 gene encoding Cx43 results in a frameshift yielding 46 aberrant amino acids after residue 259 and a shortened protein of 305 residues compared with the 382 in wild-type Cx43. This frameshift mutant (fs260) causes oculodentodigital dysplasia (ODDD) that includes the added condition of palmoplantar keratoderma. When expressed in a variety of cell lines, the fs260 mutant was typically localized to the endoplasmic reticulum and other intracellular compartments. The fs260 mutant, but not the G138R ODDD-linked Cx43 mutant or a Cx43 mutant truncated at residue 259 (T259), reduced the number of apparent gap junction plaques formed from endogenous Cx43 in normal rat kidney cells or keratinocytes. Interestingly, mutation of a putative FF endoplasmic reticulum retention motif encoded within the 46 aberrant amino acid domain failed to restore efficient assembly of the fs260 mutant into gap junctions. Dual whole cell patch-clamp recording revealed that fs260-expressing N2A cells exerted severely reduced electrical coupling in comparison to wild-type Cx43 or the T259 mutant, whereas single patch capacitance recordings showed that fs260 could also dominantly inhibit the function of wild-type Cx43. Co-expression studies further revealed that the dominant negative effect of fs260 on wild-type Cx43 was dose-dependent, and at a predicted 1:1 expression ratio the fs260 mutant reduced wild-type Cx43-mediated gap junctional conductance by over 60%. These results suggest that the 46 aberrant amino acid residues associated with the frameshift mutant are, at least in part, responsible for the manifestation of palmoplantar keratoderma symptoms.


Interferon-alpha activates multiple STAT signals and down-regulates c-Met in primary human hepatocytes.
Interferon (IFN)-alpha therapy is currently the primary choice for viral hepatitis and a promising treatment for hepatocellular carcinoma (HCC). Primary mouse and rat hepatocytes respond poorly to IFN-alpha stimulation. Thus, it is very important to examine the IFN-alpha signal pathway in primary human hepatocytes. The IFN-alpha-activated signals and genes in primary human hepatocytes and hepatoma cells were examined by Western blotting and microarray analyses. Primary human hepatocytes respond very well to IFN-alpha stimulation as shown by activation of multiple signal transducer and activator of transcription factor (STAT) 1, 2, 3, 5, and multiple genes. The differential response to IFN-alpha stimulation in primary human and mouse hepatocytes may be caused by expression of predominant functional IFN-alpha receptor 2c (IFNAR2c) in primary human hepatocytes vs. expression of predominant inhibitory IFNAR2a in mouse hepatocytes. Microarray analyses of primary human hepatocytes show that IFN-alpha up-regulates about 44 genes by over 2-fold and down-regulates about 9 genes by 50%. The up-regulated genes include a variety of antiviral and tumor suppressors/proapoptotic genes. The down-regulated genes include c-myc and c-Met, the hepatocyte growth factor (HGF) receptor. Down-regulation of c-Met is caused by IFN-alpha suppression of the c-Met promoter through down-regulation of Sp1 binding and results in attenuation of HGF-induced signals and cell proliferation. IFN-alpha directly targets human hepatocytes, followed by activation of multiple STATs and regulation of a wide variety of genes, which may contribute to the antiviral and antitumor activities of IFN-alpha in human liver.


[The provision and perioperative transfusion of erythrocyte concentrates].
A previously used order schedule for procuring blood concentrates was reviewed, as the need for blood had changed due to the application of blood saving methods. Furthermore, the indication to blood transfusion has been reduced as a result of the potential danger of infection with AIDS. In 1035 patients, we compared the number of preoperatively ordered with the number of perioperatively transfused units of blood. In addition, we documented the factors which lead to increased blood ordering. The relation between ordered and transfused units of blood in total was 596/97 (Urology: 392/78, Gynaecology: 179/18, ENT and Ophthalmology: 25/1). Anticoagulation, anaemia, secondary illnesses and specific surgical problems have been identified as factors leading to increased blood ordering. As to many units of blood are being crossmatched without being transfused, we developed a revised blood ordering schedule. According to this schedule for certain surgical procedures typing and screening only should be done rather than crossmatching.


Lifestyle modification program decreases pregnancy weight gain in obese women.
We assessed the impact of a lifestyle modification program (LMP) on weight gain in pregnancy and evaluated its effect on adverse pregnancy outcomes. A prospective matched controlled study design was employed. Inclusion criteria consisted of pregnant women with singleton gestations and body mass index (BMI) > or = 30 without underlying medical conditions. In the study group, patients were prospectively enrolled in the first trimester. Through the LMP, women participated in a comprehensive program on nutrition, exercise, and weight control in pregnancy. The control group consisted of matched patients by starting BMI, parity, and socioeconomic status during the same study period cared for at the same institution. The primary outcome evaluated was weight gain in pregnancy. Adverse pregnancy outcomes including preeclampsia, gestational diabetes, gestational hypertension, and various postpartum complications were evaluated between the two groups. Statistical analysis was performed using nonparametric methods, with p < 0.5 considered significant. Twenty-eight patients were recruited for this study and underwent the intervention. Patient compliance with the LMP was 75%, two patients were lost to follow-up, one had a preterm delivery, and four had incomplete data due to various reasons. Twenty-one patients completed their pregnancy and were available for analysis. Twenty matched control patients were identified for analysis. Study participants gained significantly less weight in their pregnancies when compared with controls (mean weight gain 17 versus 34 pounds, respectively; p = 0.008). Secondary outcomes of preeclampsia, gestational diabetes, cesarean section, as well as infant birth weight did not significantly differ between the groups. This study suggests an effective method of reducing prenatal weight gain in the obese population. This type of comprehensive intervention could be an important, cost-effective risk-reduction strategy.


Zoledronic acid: an unending tale for an antiresorptive agent.
Therapy with zoledronic acid (ZOL) is a standard for patients with malignant bone disease. However, the antitumor effects of ZOL are demonstrated only in the preclinical setting and, therefore, they require pharmacological and molecular optimization. ZOL blocks protein isoprenylation, a pleiotropic and basic biochemical process within the cells regulating the activity of several proteins involved in survival and proliferating pathways. The present review will cover different areas of research from pharmacology to cell biology and molecular biology regarding both direct and indirect anticancer effects of ZOL in humans. The challenge in demonstrating the clinical activity of ZOL is mainly based on its pharmacokinetic properties, which are discussed in detail. New data about the modes to overcome the limitations of ZOL and the following therapeutic strategies to implement its anticancer activity are described: i) development of pharmacological combinations; ii) finding of new molecular targets of ZOL; and iii) development of new pharmacological formulations of ZOL. In conclusion, ZOL has still underestimated anticancer properties that require further investigation in both preclinical and clinical settings.


Hairless descendants of Mexican hairless dogs: an experimental model for studying hypertrophic scars.
In previous studies, the author noticed hypertrophic scar formation in hairless dogs. The purpose of this study was to evaluate both grossly and histopathologically hypertrophic scars in hairless dogs to explore any similarities with hypertrophic scars in humans and to introduce these dogs as an appropriate model for further investigation on hypertrophic scarring. Full-thickness wounds were made on the dorsolumbar skin of hairless dogs. Hypertrophic scarring was examined with three methods: clinical observations, dihydroxyphenylalanine (DOPA)-positive melanocytes, and skin histopathology. Hairless dogs clinically developed the formation of hyperpigmented and hypertrophic scars that did not extend beyond the original wound margins. In hypertrophic scars of hairless dogs, the split epidermal sheet showed an increased number of DOPA-positive melanocytes with well-developed dendrites exhibiting activated melanocytes. There were very few DOPA-positive melanocytes in the repaired skin of haired dogs. Histopathologic examinations demonstrated that hypertrophic scars were fully reepithelialized and granulation tissue formation was accompanied by inflammatory cell infiltration. There was remodeling of thick collagens and fine elastic fibers in the course of hypertrophic scar formation. Experimental hypertrophic scars produced in hairless dogs have morphologic properties similar to those of human hypertrophic scars.


[Risks of autologous blood transfusion].
Underutilization of autologous blood for transfusion is widespread. Our results suggest that this problem could only gradually be corrected, as some patients show insufficient erythropoietic recovery after a predeposit autologous donation.


The Connecticut Center of Excellence for Eliminating Health Disparities among Latinos (CEHDL).
CEHDL's mission is to contribute to the elimination of health disparities among Latino(a)s through the formation of human resources, community-based research, and culturally appropriate outreach/extension. CEHDL is structured as a consortium led by the University of Connecticut (UConn) in close partnership with the Hispanic Health Council (HHC), a community health agency located in inner-city Hartford, and Hartford Hospital (HH). Demonstrating best practice and culturally skilled, evidence-based outreach, and bringing the best of academic, community, and health institutions to socioeconomically disadvantaged communities, CEHDL fosters scientific-community interactions and supports training of undergraduate, graduate, and medical students. Building capacity in other agencies is one method through which CEDHDL seeks to accomplish its goals. Thus far, CEHDL has made substantial progress demonstrating that interdisciplinary community-academic-hospital partnerships are essential for addressing health inequities in our country.


Imaging through turbulent water using speckle reference holography.
A dramatic reduction in image distortion resulting from turbulent water has been demonstrated using speckle reference holography. Conventional photographs taken simultaneously over the same path clearly show that the holographic technique eliminates most image distortion.


Mirror movements and ipsilateral motor evoked potentials in ALS.
It can be difficult to detect upper motor neuron (UMN) involvement in early amyotrophic lateral sclerosis (ALS). The present study tested the usefulness of mirror movements (MMs), i.e., contralateral coactivation of hand muscles, as a UMN sign in ALS. Thirty-seven patients with possible, probable or definite ALS and 19 patients with suspected ALS without clinical signs of UMN involvement, 21 disease controls and 15 healthy volunteers were included. MMs were studied clinically, electromyographically and by transcranial magnetic stimulation (TMS), looking for ipsilateral motor evoked potentials (IMEPs). MMs were observed clinically in 15 ALS patients (27%) and electromyographically in 28 (50%). IMEPs in the abductor pollicis brevis muscle following TMS were recorded in 34 (61%) of all ALS patients and in nine (47%) out of 19 patients with suspected ALS, but not in healthy controls. Central motor conduction times were prolonged in 15 ALS patients (27%). It is concluded that MMs, especially if studied by EMG and TMS (IMEPs) can essentially improve detection of UMN involvement, especially in early stages of ALS.


[Results of alginate and hypertonic solution in wound healing of head and neck cancers].
In head and neck cancers, complications such as pharyngostoma, orostoma, or fistula can present following surgery or surgery after irradiation. To report the results of the association of two sea-derived products (Sinomarin and Algosteril) compared to normal saline for the management of healing. This is retrospective study (1998-2000). Thirty patients were included in group 1 (Sinomarin-Algosteril) and 17 patients in group 2 (normal saline). The average age was respectively 60 and 56.2. Patients in both groups presented with advanced head and neck carcinoma (T3-T4) and healing complications. The delay of healing was respectively of 33.53 and 72.94 days. Results were statistically significant (Student T-test P < 0.05). Our results show the efficiency (p < 0.05) of the Sinomarin Algosteril association in head and neck cancer healing complications.


Randomised clinical trial: the effects of a multispecies probiotic vs. placebo on innate immune function, bacterial translocation and gut permeability in patients with cirrhosis.
Probiotics may correct intestinal dysbiosis and proinflammatory conditions in patients with liver cirrhosis. To test the effects of a multispecies probiotic on innate immune function, bacterial translocation and gut permeability. In a randomised, double blind, placebo-controlled study, stable cirrhotic out-patients either received a daily dose of a probiotic powder containing eight different bacterial strains (Ecologic Barrier, Winclove, Amsterdam, The Netherlands) (n = 44) or a placebo (n = 36) for 6 months and were followed up for another 6 months. We found a significant but subclinical increase in neutrophil resting burst (2.6-3.2%, P = 0.0134) and neopterin levels (7.7-8.4 nmol/L, P = 0.001) with probiotics but not with placebo. Probiotic supplementation did not have a significant influence on neutrophil phagocytosis, endotoxin load, gut permeability or inflammatory markers. Ten severe infections occurred in total; one during intervention in the placebo group, and five and four after the intervention has ended in the probiotic and placebo group, respectively. Liver function showed some improvement with probiotics but not with placebo. Probiotic supplementation significantly increased serum neopterin levels and the production of reactive oxygen species by neutrophils. These findings might explain the beneficial effects of probiotics on immune function. Furthermore, probiotic supplementation may be a well-tolerated method to maintain or even improve liver function in stable cirrhosis. However, its influence on gut barrier function and bacterial translocation in cirrhotic patients is minimal.


Versatile Electroanalytical Bioplatforms for Simultaneous Determination of Cancer-Related DNA 5-Methyl- and 5-Hydroxymethyl-Cytosines at Global and Gene-Specific Levels in Human Serum and Tissues.
This paper reports the preparation of versatile electrochemical biosensing platforms for the simple, rapid, and PCR-independent detection of the most frequent DNA methylation marks (5-methylcytosine, 5-mC, and/or 5-hydroxymethylcytosine, 5-hmC) both at global and gene-specific levels. The implemented strategies, relying on the smart coupling of immuno-magnetic beads (MBs), specific DNA probes and amperometric detection at screen-printed carbon electrodes (SPCEs), provided sensitive and selective determination of the target methylated DNAs in less than 90 min with a great reproducibility and demonstrated feasibility for the simultaneous detection of the same or different cytosine epimarks both at global level and in different loci of the same gene or in different genes. The bioplatforms were applied to determine global methylation events in paraffin-embedded colorectal tissues and specific methylation at promoters of tumor suppressor genes in genomic DNA extracted from cancer cells and paraffin-embedded colorectal tissues, and in serum without previous DNA extraction from cancer patients.


Mutagenic Activation of Glutathione Peroxidase-4: Approaches toward Rational Design of Allosteric Drugs.
Glutathione peroxidase 4 (GPX4) reduces lipid hydroperoxides in lipid membranes, effectively inhibiting iron-dependent cell death or ferroptosis. The upregulation of the enzyme by the mutations at residues D21 and D23 has been suggested to be associated with higher protein activity, which confers more protection against neurodegenerative diseases such as Alzheimer's, Parkinson's, and Huntington's diseases. Therefore, it has become an attractive target for treating and preventing neurodegenerative diseases. However, identifying means of mimicking the beneficial effects of these mutations distant from the active site constitutes a formidable challenge in moving toward therapeutics. In this study, we explore using molecular dynamics simulations to computationally map the conformational and energetic landscape of the wild-type GPX4 protein and three mutant variants to identify the allosteric networks of the enzyme. We present the conformational dynamic profile providing the desired signature behavior of the enzyme. We also discuss the implications of these findings for drug design efforts.


Hyperbaric oxygen therapy improves neurogenesis and brain blood supply in piriform cortex in rats with vascular dementia.
Neural stem cell treatment and neurogenesis stimulation have gained attention as potential treatments for vascular dementia (VD). Currently, research mainly focuses on neurogenesis occurring in the sub-ventricular zone and dentate gyrus, while the research of the piriform cortex (Pir) is limited. Few results showed that weak neurogenesis exists in the Pir of adult rats. Since neurogenesis occurs in the Pir and is closely related to cognitive function, this study addressed the question of whether neurogenesis occurs in the Pir of an animal with the VD. PRIMARY OBJECTIVE AND HYPOTHESIS: This study investigated the effects of hyperbaric oxygen therapy on brain blood supply and neurogenesis in the piriform cortex (Pir) of rats with VD. Compared to non-VD control rats (NC), rats with VD showed reduced rCBF, increased rCBV and slower MTT in the Pir. However, following hyperbaric oxygen (HBO) treatment in VD rats, rCBF increased, rCBV decreased and MTT increased. To determine whether the restoration in brain blood supply was associated with increased neurogenesis, immunohistochemical detection of nestin and doublecortin (DCX) was used. In the Pir of both normal and VD rats, nestin positive cells were localized to layer II (superficial cellular layer) and layer III (deep cellular layer). Nestin expression was increased in Pir cells in VD rats and was even more intensely expressed after the HBO treatment. DCX positive cells were mostly located in layer II from amygdaline fissure to rhinal fissure. These results suggest that HBO therapy can improve the blood supply and promote the neurogenesis in the Pir of adult rats with the VD.


Radiocaesium in Cortinarius spp. mushrooms in the regions of the Reggio Emilia in Italy and Pomerania in Poland.
Activity concentrations of 134Cs and 137Cs have been determined in 23 species of mushrooms of the genus Cortinarius (59 individual samples) collected from the Reggio Emilia in Italy 1992-1999 and in 4 species (16 composite samples and 413 individuals) from the Pomerania region in Poland from 1996 to 2015. Across all the Cortinarius species from the Reggio Emilia, the activity concentrations were relatively high in Cortinarius alboviolaceus, Cortinarius duracinus, Cortinarius orellanus, Cortinarius rapaceus, and Cortinarius subannulatus, in which 137Cs was at 10,000 ~ 100,000 Bq kg-1 dry biomass (db) in 1994. Smaller activity concentrations were found in Cortinarius bivelus, Cortinarius bulliardii, Cortinarius cotoneus, Cortinarius largus, Cortinarius lividoviolaceus, Cortinarius purpureus, Cortinarius rufo-olivaceus, Cortinarius torvus, and Cortinarius venetus with levels at 1000 ~ 6000 Bq kg-1 db from 1992 to 1994, and further in Cortinarius anserinus, Cortinarius auroturbinatus, C. largus, Cortinarius praestans, Cortinarius purpurascens, Cortinarius scaurus, Cortinarius sebaceous, Cortinarius talus, and Cortinarius variecolor with activity concentrations at 100 ~ 600 Bq kg-1 db in 1994. All the data were calculated for dehydrated fungal material corrected back to the exact date samples of collection. The greatest activity concentrations of 137Cs both in Italy (1992-1999) and Poland (1996-2010) were found in the popular Cortinarius caperatus, confirming its very high capacity of radiocaesium accumulation. Besides 137Cs, the isotope 134Cs was detected in some species from the Reggio Emilia. An average calculated ratio of activities of 134Cs to 137Cs referenced to 1986 was equal to 0.38 in mushrooms from the Reggio Emilia, and this value slightly differ from that specific for Chernobyl fallout, which was 0.54. It was calculated that 137Cs originating from Chernobyl accident constituted about 68 % of the total activity concentration of the isotope in Reggio Emilia in 1986, while as much as 32 % of 137Cs in mushrooms were from the global fallout from nuclear bomb testing.


First-trimester screening for trisomies by cfDNA testing of maternal blood in singleton and twin pregnancies: factors affecting test failure.
To examine factors affecting the rate of failure to obtain a result from cell-free DNA (cfDNA) testing of maternal blood for fetal trisomies 21, 18 and 13 in singleton and twin pregnancies in the first trimester. This was a prospective study of 23 495 singleton and 928 twin pregnancies undergoing screening for fetal trisomy by targeted cfDNA testing at 10 + 0 to 14 + 1 weeks' gestation. Multivariate logistic regression analysis was used to determine significant predictors of failure to obtain a result after first sampling. There was no result from cfDNA testing after first sampling in 3.4% (798/23 495) of singletons, 11.3% (91/806) of dichorionic twins and 4.9% (6/122) of monochorionic twins. Multivariate logistic regression analysis demonstrated that the risk of test failure, first, increased with increasing maternal age (odds ratio (OR), 1.02; 95% CI, 1.01-1.04) and weight (OR, 1.05; 95% CI, 1.04-1.05), decreasing gestational age (OR, 0.85; 95% CI, 0.79-0.91), serum pregnancy-associated plasma protein-A (PAPP-A) multiples of the median (MoM) (OR, 0.56; 95% CI, 0.49-0.65) and free β-human chorionic gonadotropin (β-hCG) MoM (OR, 0.67; 95% CI, 0.60-0.74), second, was higher in women of black (OR, 1.72; 95% CI, 1.33-2.20) and South Asian (OR, 1.99; 95% CI, 1.56-2.52) than those of white racial origin, in dichorionic twin than in singleton pregnancy (OR, 1.75; 95% CI, 1.34-2.26) and in pregnancies conceived by in-vitro fertilization than in those conceived naturally (OR, 3.82; 95% CI, 3.19-4.55) and, third, was lower in parous than in nulliparous women (OR, 0.63; 95% CI, 0.55-0.74). Maternal age, weight, racial origin and parity, gestational age, dichorionicity, method of conception and serum levels of free β-hCG and PAPP-A are independent predictors of cfDNA test failure. The risk of test failure is higher in dichorionic twin than in singleton pregnancies, mainly because a higher proportion of twins are conceived by in-vitro fertilization and more of the women are nulliparous. Copyright © 2019 ISUOG. Published by John Wiley & Sons Ltd.