Diminished Th1-type cytokine production in gastric mucosa T-lymphocytes after H. pylori eradication in duodenal ulcer patients.
Helicobacter pylori infects an estimated 50% of the world population, however only a small proportion of individuals develop clinical symptoms of gastritis, peptic ulceration or gastric cancer. The variations in disease presentation may be due to differences in bacterial virulence and/or immune response to the pathogen. In the previous study we reported an increased expression of the IL-2 receptor in duodenal ulcer (DU) patients infected with H. pylori. This study examined intracellular lymphokine production in gastric mucosa infiltrating T lymphocytes in DU patients before and after H. pylori eradication. T lymphocytes were isolated from gastric mucosa biopsies by using mechanical and enzymatic tissue desegregation. Ficoll-purified lymphocytes were incubated with monoclonal antibodies and analysed by using 3-colour flow cytometry analysis for intracellular interferon gamma (IFNgamma) and interleukin 4 (IL-4) expression in order to define Th1 and Th2 cell population. We demonstrated a significant decrease in the proportion of Th1 cells infiltrating gastric mucosa 6 and 12 months after H. pylori eradication. Our results suggest the importance of the local immune response in the development of H. pylori related gastritis.


Phosphorylation of human gp130 at Ser-782 adjacent to the Di-leucine internalization motif. Effects on expression and signaling.
The receptor for leukemia inhibitory factor (LIF) consists of two polypeptides, the LIF receptor and gp130. Agonist stimulation has been shown previously to cause phosphorylation of gp130 on serine, threonine, and tyrosine residues. We found that gp130 fusion proteins were phosphorylated exclusively on Ser-782 by LIF- and growth factor-stimulated 3T3-L1 cell extracts. Ser-780 was required for phosphorylation of Ser-782 but was not itself phosphorylated. Ser-782 is located immediately N-terminal to the di-leucine motif of gp130, which regulates internalization of the receptor. Transient expression of chimeric granulocyte colony-stimulating factor receptor (G-CSFR)-gp130(S782A) receptors resulted in increased cell surface expression in COS-7 cells and increased ability to induce vasoactive intestinal peptide gene expression in IMR-32 neuroblastoma cells when compared with expression of chimeric receptors containing wild-type gp130 cytoplasmic domains. These results identify Ser-782 as the major phosphorylated serine residue in human gp130 and indicate that this site regulates cell surface expression of the receptor polypeptide.


Peak oxygen uptake measured during a perceptually-regulated exercise test is reliable in community-based manual wheelchair users.
This study compares test-retest reliability and peak exercise responses from ramp-incremented (RAMP) and maximal perceptually-regulated (PRETmax) exercise tests during arm crank exercise in individuals reliant on manual wheelchair propulsion (MWP). Ten untrained participants completed four trials over 2-weeks (two RAMP (0-40 W + 5-10 W · min-1) trials and two PRETmax. PRETmax consisted of five, 2-min stages performed at Ratings of Perceived Exertion (RPE) 11, 13, 15, 17 and 20). Participants freely changed the power output to match the required RPE. Gas exchange variables, heart rate, power output, RPE and affect were determined throughout trials. The V̇O2peak from RAMP (14.8 ± 5.5 ml · kg-1 · min-1) and PRETmax (13.9 ± 5.2 ml · kg-1 · min-1) trials were not different (P = 0.08). Measurement error was 1.7 and 2.2 ml · kg-1 · min-1 and coefficient of variation 5.9% and 8.1% for measuring V̇O2peak from RAMP and PRETmax, respectively. Affect was more positive at RPE 13 (P = 0.02), 15 (P = 0.01) and 17 (P = 0.01) during PRETmax. Findings suggest that PRETmax can be used to measure V̇O2peak in participants reliant on MWP and leads to a more positive affective response compared to RAMP.


Inspiring innovation in medical education.
Traditionally, changes to medical education come from the top down, an approach that potentially misses important contributions from medical students, residents, faculty and staff. In order to provide an avenue for them to bring forward their ideas for educational improvements, the University of Minnesota Medical School sponsored the "What's the Bright Idea?" contest. Through the contest, we sought to foster a culture of innovation and collaboration among faculty, staff and students. The contest included five phases: launch, idea submission, online voting, follow-up and implementation. Seventy-six ideas were submitted, and 902 people participated in the online voting. When asked in a follow-up survey whether the submitter would have developed their idea without the contest, 27% of respondents answered "no" and 18% answered "maybe." Three-fourths stated the contest stimulated networking and collaboration. Four of the recommendations are now being implemented.


Effect of n-butyrate on superinfection of Raji lymphoblastoid cell line by Epstein-Barr virus.
The effect of n-butyrate on superinfectability of virus-nonproducer Raji cells by the P3HR-1 strain of Epstein-Barr virus (EBV) was investigated. n-Butyrate is known to be a potent inducer of virus antigen synthesis in virus-producer cell lines and of cell differentiation in virus nonproducers. The drug inhibited the growth of Raji cells but did not interfere markedly with cell viability. It induced a low rate of early antigen (EA) synthesis in about 1-2% of noninfected Raji cells. While the number of superinfectable cells remained relatively constant after treatment with butyrate, an increase in antigen positivity was noted in untreated cells. This relative decrease in sensitivity to superinfection in butyrate-treated Raji cells was more pronounced in cultures that had been treated with the drug for 48 or 72 hr as compared to those treated for 24 hr. A blocking of the treated cells in the certain cell-cycle phase and their drug-induced differentiation towards plasma cells might have been involved in the phenomenon described.


Effect of adhesive matrix composition and terpinolene on indomethacin bioavailability in rats from transdermal therapeutic system.
Drug-in-adhesive matrix-type transdermal therapeutic systems for indomethacin (IND) were formulated and evaluated. Silicone and two types of polyacrylates were used as the bases of matrices. Terpinolene was used as a penetration enhancer. The physicochemical properties of matrices were determined. The bioavailability study of IND was performed in rats. The presence of IND in blood was demonstrated for each system. The calculated pharmacokinetics parameters for IND mainly depend on the solubility of IND in the adhesive layer. The positive influence of a penetration enhancer on IND bioavailability was observed only for one type of polyacrylate matrices.


Predominant cultivable flora isolated from human root surface caries plaque.
Plaque samples were obtained from tooth surfaces exhibiting typical lesions of root surface caries and were immediately cultured by a continuous anaerobic procedure. The bacterial composition of root caries flora was determined on individual samples. Representative isolates from each specimen were characterized by morphological and physiological criteria. In addition, fluorescent antibody reagents were used to confirm the identification of Streptococcus mutans and Actinomyces viscosus. The plaque samples could be divided into two groups on the basis of the presence or absence of S. mutans in the plaque. In group I plaques, S. mutans comprised 30 percent of the total cultivable flora. S. sanguis was either not found or was present in very low number. In group II plaques, S. mutans was not detected, and S. sanguis formed 48 percent of the total plaque flora. A. viscosus was the dominant organism in all plaque samples, accounting for 47 percent of the group I isolates and 41 percent of the group II isolates.


Talin requires beta-integrin, but not vinculin, for its assembly into focal adhesion-like structures in the nematode Caenorhabditis elegans.
In cultured cells, the 230-kDa protein talin is found at discrete plasma membrane foci known as focal adhesions, sites that anchor the intracellular actin cytoskeleton to the extracellular matrix. The regulated assembly of focal adhesions influences the direction of cell migrations or the reorientation of cell shapes. Biochemical studies of talin have shown that it binds to the proteins integrin, vinculin, and actin in vitro. To understand the function of talin in vivo and to correlate its in vitro and in vivo biochemical properties, various genetic approaches have been adopted. With the intention of using genetics in the study of talin, we identified a homologue to mouse talin in a genetic model system, the nematode Caenorhabditis elegans. C. elegans talin is 39% identical and 59% similar to mouse talin. In wild-type adult C. elegans, talin colocalizes with integrin, vinculin, and alpha-actinin in the focal adhesion-like structures found in the body-wall muscle. By examining the organization of talin in two different C. elegans mutant strains that do not make either beta-integrin or vinculin, we were able to determine that talin does not require vinculin for its initial organization at the membrane, but that it depends critically on the presence of integrin for its initial assembly at membrane foci.


Interactions between the intestinal microbiome and liver diseases.
The human intestine harbors a diverse community of microbes that promote metabolism and digestion in their symbiotic relationship with the host. Disturbance of its homeostasis can result in disease. We review factors that disrupt intestinal homeostasis and contribute to nonalcoholic fatty liver disease, steatohepatitis, alcoholic liver disease, and cirrhosis. Liver disease has long been associated with qualitative and quantitative (overgrowth) dysbiotic changes in the intestinal microbiota. Extrinsic factors, such as the Western diet and alcohol, contribute to these changes. Dysbiosis results in intestinal inflammation, a breakdown of the intestinal barrier, and translocation of microbial products in animal models. However, the contribution of the intestinal microbiome to liver disease goes beyond simple translocation of bacterial products that promote hepatic injury and inflammation. Microbial metabolites produced in a dysbiotic intestinal environment and host factors are equally important in the pathogenesis of liver disease. We review how the combination of liver insult and disruptions in intestinal homeostasis contribute to liver disease.


The Role of Non-Coding RNAs in Chromosomal Instability in Cancer.
Chromosomal instability (CIN) is characterized by an increased frequency of changes in chromosome structure or number and is regarded as a hallmark of cancer. CIN plays a prevalent role in tumorigenesis and cancer progression by assisting the cancer cells' phenotypic adaptation to stress, which have been tightly linked to therapy resistance and metastasis. Both CIN-inducing and CIN-repressing agents are being clinically tested for the treatment of cancer to increase CIN levels to unsustainable levels leading to cell death or to decrease CIN levels to limit the development of drug resistance, respectively. Non-coding RNAs (ncRNAs) including microRNAs and long ncRNAs (lncRNAs) have been fundamentally implicated in CIN. The miR-22, miR-26a, miR-28, and miR-186 target important checkpoint proteins involved in mediating chromosomal stability and their expression modulation has been directly related to CIN occurrence. lncRNAs derived from telomeric, centrosomal, and enhancer regions play an important role in mediating genome stability, while specific lncRNA transcripts including genomic instability inducing RNA called Ginir, P53-responsive lncRNA termed as GUARDIN, colon cancer-associated transcript 2, PCAT2, and ncRNA activated by DNA damage called NORAD have been shown to act within CIN-associated pathways. In this review, we discuss how these ncRNAs either maintain or disrupt the stability of chromosomes and how these mechanisms could be exploited for novel therapeutic approaches targeting CIN in cancer patients. SIGNIFICANCE STATEMENT: Chromosomal instability increases tumor heterogeneity and thereby assists the phenotypic adaptation of cancer cells, causing therapy resistance and metastasis. Several microRNAs and long non-coding RNAs that have been causally linked to chromosomal instability could represent novel therapeutic targets. Understanding the role of non-coding RNAs in regulating different genes involved in driving chromosomal instability will give insights into how non-coding RNAs can be utilized toward modifying chemotherapeutic regimens in different cancers.


Characterization of the Francisella tularensis subsp. novicida type IV pilus.
Francisella tularensis causes the disease tularaemia. Type IV pili (Tfp) genes are present in the genomes of all F. tularensis subspecies. We show that the wild-type F. tularensis subsp. novicida expresses pilus fibres on its surface, and mutations in the Tfp genes pilF and pilT disrupt pilus biogenesis. Mutations in other Tfp genes (pilQ and pilG) do not eliminate pilus expression. A mutation in pilE4 eliminates pilus expression, whereas mutations in the other pilin subunits pilE1-3 and pilE5 do not, suggesting that pilE4 is the major pilus structural subunit. The virulence regulator MglA is required for pilus expression, and it regulates the transcription of a putative Tfp glycosylation gene (FTN0431). However, MglA does not regulate transcription of pilF, pilT or pilE4, and a strain lacking FTN0431 still expresses pili; thus, it is unclear how MglA regulates pilus expression. Only pilF was also required for protein secretion, while pilE4 and pilT were not, indicating that there is very little overlap of the protein secretion/Tfp functions of the pil genes. The protein secretion component pilE1 was more important for in vitro intramacrophage growth and mouse virulence than the Tfp component pilE4. Our results provide the first genetic characterization of the novel Tfp system of F. tularensis.


Activation of the maize anthocyanin gene a2 is mediated by an element conserved in many anthocyanin promoters.
Two transcription factors, C1 (a Myb-domain protein) and B (a basic-helix-loop-helix protein), mediate transcriptional activation of the anthocyanin-biosynthetic genes of maize (Zea mays). To begin to assess the mechanism of activation, the sequences required for C1- and B-mediated induction have been determined for the a2 promoter, which encodes an anthocyanin-biosynthetic enzyme. Analysis of a series of 7- to 13-base-pair substitutions revealed two regions crucial for activation. One region, centered at -99, contained a C1-binding site that abolished C1 binding. The other crucial region was adjacent, centered at -91. C1 binding was not detected at this site, and mutation of this site did not prevent C1 binding at -99. An oligonucleotide dimer containing these two crucial elements was sufficient for C1 and B activation of a heterologous promoter. These data suggest that activation of the anthocyanin genes involves C1 and another factor binding at closely adjacent sites. Mutating a previously postulated anthocyanin consensus sequence within a2 did not significantly reduce activation by C1 and B. However, sequence comparisons of the crucial a2 regions with sequences important for C1- and B-mediated activation in two other anthocyanin promoters led to a revised consensus element shared by these promoters.


Identification of pneumococcal surface protein A as a lactoferrin-binding protein of Streptococcus pneumoniae.
Lactoferrin (Lf), an iron-sequestering glycoprotein, predominates in mucosal secretions, where the level of free extracellular iron (10(-18) M) is not sufficient for bacterial growth. This represents a mechanism of resistance to bacterial infections by prevention of colonization of the host by pathogens. In this study we were able to show that Streptococcus pneumoniae specifically recognizes and binds the iron carrier protein human Lf (hLf). Pretreatment of pneumococci with proteases reduced hLf binding significantly, indicating that the hLf receptor is proteinaceous. Binding assays performed with 63 clinical isolates belonging to different serotypes showed that 88% of the tested isolates interacted with hLf. Scatchard analysis showed the existence of two hLf-binding proteins with dissociation constants of 5.7 x 10(-8) and 2.74 x 10(-7) M. The receptors were purified by affinity chromatography, and internal sequence analysis revealed that one of the S. pneumoniae proteins was homologous to pneumococcal surface protein A (PspA). The function of PspA as an hLf-binding protein was confirmed by the ability of purified PspA to bind hLf and to competitively inhibit hLf binding to pneumococci. S. pneumoniae may use the hLf-PspA interaction to overcome the iron limitation at mucosal surfaces, and this might represent a potential virulence mechanism.


Word and tone working memory deficits in schizophrenia.
Verbal memory deficits have been reported in many studies of patients with schizophrenia. We evaluated the specificity of these deficits by comparing patients and control subjects on several verbal and nonverbal auditory memory tests. Performance of stable, medicated outpatients with DSM-III-R diagnoses of schizophrenia (N = 38) was compared with that of healthy subjects (N = 39) on a word list immediate recall task, tone delayed discrimination tasks, and word and tone serial position tasks. Before memory testing, patients were divided into 2 groups based on their ability to perform normally on a screening test requiring pitch discrimination and sustained attention. The nonverbal tests were more difficult for control subjects than the verbal tests. Despite this, patients who performed normally on the screening test of perception and attention performed normally on both nonverbal tests but had highly significant deficits on both verbal tests (P<.001 and P = .02). Patients who performed poorly on the screening test had highly significant performance deficits on all the memory tests. One subgroup of patients with schizophrenia has a selective deficit in verbal memory despite normal motivation, attention, and general perceptual function. Another group has deficits in multiple aspects of cognitive function suggestive of failure in early stages of information processing.


The importance of contacts and embrasures and their effect on the periodontium.
Operative and restorative dentistry are challenging disciplines of dentistry. When esthetics and periodontics are added to the equation, it becomes that much more difficult for any operator to restore teeth in an ideal manner. In order to be successful and have reasonable occlusal harmony, periodontal health, and beautiful esthetics, knowledge of all of these disciplines is necessary no matter what kind of material is being placed. Continuing education, care, skill, practice, a desire for excellence, and, most of all, patience will allow any dentist to accomplish these goals.


An in vitro study of lipid preference in whaleworm (Anisakis simplex, Nematoda, Ascaridoidea, Anisakidae) third-stage larvae.
The behavioural response of nematodes to chemical stimuli has been extensively investigated in some free-living and plant parasitic species. However, in animal parasitic species, little is yet known, particularly in regards to marine forms such as the whaleworm (Anisakis simplex). Previous studies showed that A. simplex L3-larvae tend to prefer fish tissue with high lipid content. The intention of this study was to investigate the behaviour of A. simplex L3 in response to different concentrations of fish lipid in further detail. This was done by an in vitro study based on larvae from cod (Gadus morhua). Ten larvae were placed in each of the culture containers containing agar that was separated into three segments of equal size. Three categories of agar were used containing 0, 2 and 7% cod liver oil. A total of 900 larvae were included. The study consisted of three parts: The purpose of experiment I was to establish whether different lipid concentrations influenced the migration pattern at all. Experiment II was intended to examine whether A. simplex L3-larvae were able to actively search for lipids. Experiment III was set up to analyse the short-distance dispersion of the L3-larvae. Experiment I indicated that the L3-larvae move randomly but do not stop randomly since the tendency to move out of the start area was inversely correlated with lipid concentration. Experiment II indicates that the larvae are almost unable to select areas of high lipid concentrations when more than a few centimetres away. Experiment III showed that the L3-larvae prefer high-fat content and can seek it out over short distances.


Interstitial Eosinophilic Aggregates and Kidney Outcome in Patients with CKD.
Interstitial eosinophilic aggregates are observed in various kidney diseases, but their clinical implications remain unknown. We assessed the association between interstitial eosinophilic aggregates and kidney outcomes and further analyzed the association between blood eosinophil count, as a surrogate for interstitial eosinophilic aggregates, and the risk of kidney failure in patients with advanced CKD. We analyzed datasets from two retrospective cohort studies: ( 1 ) the kidney biopsy cohort including 563 patients who underwent native kidney biopsy at Osaka University Hospital between 2009 and 2021 and ( 2 ) the retrospective CKD cohort including 2877 patients with an eGFR of 10-60 ml/min per 1.73 m 2 referred to the nephrology outpatient center at Osaka University Hospital between 2005 and 2018. Interstitial eosinophilic aggregates were defined as ≥5 interstitial eosinophils in the high-power field on hematoxylin and eosin staining. This study outcome was initiation of KRT or ≥40% decline in eGFR. In the kidney biopsy cohort, interstitial eosinophilic aggregates were found in 17% of patients, most frequently in those with diabetic nephropathy (50%). Interstitial eosinophilic aggregates were associated with a higher rate of the composite kidney outcome after adjustment for clinical and histological variables (hazard ratio, 3.61; 95% confidence interval, 2.47 to 5.29; P < 0.001). LASSO revealed that blood eosinophil count was the strongest predictor of interstitial eosinophilic aggregates. In the retrospective CKD cohort, higher baseline and time-updated blood eosinophil counts were significantly associated with a higher rate of KRT initiation in Cox proportional hazards models and marginal structural models. Interstitial eosinophilic aggregates were associated with a higher risk of a composite of KRT initiation or ≥40% decline in eGFR. This article contains a podcast at https://dts.podtrac.com/redirect.mp3/www.asn-online.org/media/podcast/CJASN/2023_12_08_CJN0000000000000277.mp3.


Dexras1 Deletion and Iron Chelation Promote Neuroprotection in Experimental Optic Neuritis.
Dysregulation of iron metabolism, and resultant cytotoxicity, has been implicated in the pathogenesis of multiple sclerosis (MS) and other neurodegenerative processes. Iron accumulation promotes cytotoxicity through various mechanisms including oxidative stress and glutamate toxicity, and occurs in both MS patients and in the experimental autoimmune encephalomyelitis (EAE) model of MS. Divalent Metal Transporter1, a major iron importer in cells, is stimulated by signaling of Dexras1, a small G protein member of the Ras family. Dexras1 is activated by S-nitrosylation by nitric oxide (NO) produced by either inducible nitric oxide synthase in activated microglia/macrophages or neuronal nitric oxide synthase in neurons. Here we show Dexras1 exacerbates oxidative stress-induced neurodegeneration in experimental optic neuritis, an inflammatory demyelinating optic nerve condition that occurs in MS and EAE. Dexras1 deletion, as well as treatment with the iron chelator deferiprone, preserves vision and attenuates retinal ganglion cell (RGC) and axonal loss during EAE optic neuritis. These results suggest that iron entry triggered by NO-activated Dexras1 signaling is a potential mechanism of neuronal death in experimental optic neuritis. The current data suggest modulation of Dexras1 signaling and iron chelation are potential novel treatment strategies for optic neuritis and MS, and possibly other optic neuropathies as well.


HLA-haploidentical transplantation with regulatory and conventional T-cell adoptive immunotherapy prevents acute leukemia relapse.
Posttransplant relapse is still the major cause of treatment failure in high-risk acute leukemia. Attempts to manipulate alloreactive T cells to spare normal cells while killing leukemic cells have been unsuccessful. In HLA-haploidentical transplantation, we reported that donor-derived T regulatory cells (Tregs), coinfused with conventional T cells (Tcons), protected recipients against graft-versus-host disease (GVHD). The present phase 2 study investigated whether Treg-Tcon adoptive immunotherapy prevents posttransplant leukemia relapse. Forty-three adults with high-risk acute leukemia (acute myeloid leukemia 33; acute lymphoblastic leukemia 10) were conditioned with a total body irradiation-based regimen. Grafts included CD34(+) cells (mean 9.7 × 10(6)/kg), Tregs (mean 2.5 × 10(6)/kg), and Tcons (mean 1.1 × 10(6)/kg). No posttransplant immunosuppression was given. Ninety-five percent of patients achieved full-donor type engraftment and 15% developed ≥grade 2 acute GVHD. The probability of disease-free survival was 0.56 at a median follow-up of 46 months. The very low cumulative incidence of relapse (0.05) was significantly better than in historical controls. These results demonstrate the immunosuppressive potential of Tregs can be used to suppress GVHD without loss of the benefits of graft-versus-leukemia (GVL) activity. Humanized murine models provided insights into the mechanisms underlying separation of GVL from GVHD, suggesting the GVL effect is due to largely unopposed Tcon alloantigen recognition in bone marrow.


An algorithm for Morphological Phylogenetic Analysis with Inapplicable Data.
Morphological data play a key role in the inference of biological relationships and evolutionary history and are essential for the interpretation of the fossil record. The hierarchical interdependence of many morphological characters, however, complicates phylogenetic analysis. In particular, many characters only apply to a subset of terminal taxa. The widely used "reductive coding" approach treats taxa in which a character is inapplicable as though the character's state is simply missing (unknown). This approach has long been known to create spurious tree length estimates on certain topologies, potentially leading to erroneous results in phylogenetic searches-but pratical solutions have yet to be proposed and implemented. Here, we present a single-character algorithm for reconstructing ancestral states in reductively coded data sets, following the theoretical guideline of minimizing homoplasy over all characters. Our algorithm uses up to three traversals to score a tree, and a fourth to fully resolve final states at each node within the tree. We use explicit criteria to resolve ambiguity in applicable/inapplicable dichotomies, and to optimize missing data. So that it can be applied to single characters, the algorithm employs local optimization; as such, the method provides a fast but approximate inference of ancestral states and tree score. The application of our method to published morphological data sets indicates that, compared to traditional methods, it identifies different trees as "optimal." As such, the use of our algorithm to handle inapplicable data may significantly alter the outcome of tree searches, modifying the inferred placement of living and fossil taxa and potentially leading to major differences in reconstructions of evolutionary history.


Adding, defining and measuring value.
Suppliers to the health care industry are changing relationships with their customers by forging partnerships that blur the boundaries between them. Here, representatives from several suppliers discuss what programs and services the market has driven them to offer.


Living in the here and now: interrelationships between impulsivity, mindfulness, and alcohol misuse.
Impulsivity and mindfulness both emphasize orientation to the present, and both have been linked to alcohol misuse, but the relationship between the two is not clearly understood. The objectives of this study are to examine the relationships between elements of impulsivity and mindfulness and to examine both variables in relation to alcohol misuse. Young adults (N = 116) were assessed for alcohol use, mindfulness, and impulsivity using psychometrically validated measures. Numerous significant associations were present among the facets of impulsivity and mindfulness. All impulsivity facets and three facets of mindfulness were related to alcohol consumption and adverse consequences from drinking. After controlling for other variables, only the impulsivity domains of Negative Urgency (NU), Positive Urgency, and delay discounting were significantly related to alcohol consumption and only Lack of Premeditation and NU were significantly associated with drinking-related consequences. There was considerable overlap between some elements of impulsivity and mindfulness while the overlap was negligible for other facets. The associations between mindfulness and alcohol misuse were entirely a function of impulsivity. In particular, acting on impulses while experiencing a negative affect was significantly associated with level of alcohol consumption and level of alcohol-related risk. Steep discounting of future rewards was associated with alcohol consumption while poor premeditation was associated with adverse drinking consequences. These findings illustrate the importance of jointly studying impulsivity when examining mindfulness traits.


Targeting of hepatitis C virus core protein for MHC I or MHC II presentation does not enhance induction of immune responses to DNA vaccination.
We analyzed different vaccine approaches aimed at enhancing CD4(+)- and CD8(+)-dependent responses against hepatitis C virus (HCV) core antigen. Specific DNA vectors expressing various forms of the core in fusion with the ubiquitin or the lysosome-associated membrane protein (LAMP) were generated. These expressed the full-length wildtype core; the full-length core expressed as a covalent fusion with the ubiquitin; the full-length core expressed as a noncovalent fusion with the ubiquitin and containing a N-stabilizing or N-destabilizing residue; and the full-length core expressed as a fusion with the LAMP sequence. In vitro expression levels of the different plasmids differed by as much as tenfold. After injection into mice, none of the plasmids yielded a detectable antibody response, whereas core-specific cytotoxic T-lymphocyte (CTL) activity could be observed with all plasmids as long as 21 weeks postimmunization. No increase in CTL activity (ranging from 7% to 34% specific lysis) was observed with the ubiquitin-fusion-expressed core antigens compared with the wildtype core. The lowest CTL activity (< 5% specific lysis) was observed with the LAMP fusion. This vector was nonetheless unable to induce a detectable proliferative response. Screening of 10 different putative CTL peptide epitopes failed to reveal newly targeted epitopes when the core-fusion plasmids were used compared with the wildtype core-expressing plasmid. These data underline the difficulty in optimizing anti-core cellular immune response using molecular targeting strategies in DNA-based vaccination.


Architecture of the medial gastrocnemius in children with spastic diplegia.
Ultrasound images were obtained of the medial gastrocnemius at different ankle joint positions with the knee extended. Fascicle length and deep fascicle angle were measured in five normally developing adults (mean age 33 years, age range 24 to 36 years) and in five normally developing children (mean age 7.8 years, age range 7 to 11 years), and in seven children with spastic diplegia (mean age 10 years, age range 6 to 13 years). These architectural variables were similar in the groups of normally developing adults and children. Importantly, no statistical difference could be found between the normally developing children and those with diplegia for fascicle length. Deep fascicle angles were reduced significantly in the clinical group at a particular ankle joint angle but not at the resting angles. The difference in deep fascicle angles is explained as a function of resting muscle length and is not attributed any clinical importance. Our results do not explain the structural origin of muscle contracture explicitly. However, they do indicate that most of the fixed shortness in the medial gastrocnemii of ambulant children with spastic diplegia is not due to reduced muscle fibre length. We suggest that muscle contracture may be better explained in terms of shortness of the aponeuroses of pennate muscles, such as the medial gastrocnemius, through reduced muscle fibre diameter.


Prevalence of Helicobacter pylori in peptic ulcer disease in a Singapore hospital.
High Helicobacter pylori (H. pylori) prevalence is well documented among peptic ulcer patients. However, there have been recent reports of declining H. pylori infection rates in developed countries. Based on previous local data in a different hospital, H. pylori prevalence was 66% in gastric ulcer, 86% in duodenal ulcer and 75% in combined gastric and duodenal ulcers. Our present study aims to review the recent H. pylori prevalence in peptic ulcer patients in a Singapore hospital. H. pylori infection rate in relation to sex, age, non-steroidal anti-inflammatory drug (NSAID) use and race were also examined. Over a 6-month period, patients diagnosed with peptic ulcer by oesophagogastroduodenoscopy were selected. Relevant information was obtained from case notes retrieved from the Medical Records Office. H. pylori status was assessed by rapid urease test (CLOtest) and/or histology. Exclusion criteria were(1) history of peptic ulcer(2), active bleeding(3), cancer or(4) recent use of antibiotics or proton pump inhibitors. 107 peptic ulcer patients were selected; 53 gastric ulcer, 47 duodenal ulcer and 7 with combined gastric and duodenal ulcers. Overall H. pylori prevalence was 67.9%(36/53), 85.1%(40/47) and 85.7%(6/7) respectively. Except for the gender variable in gastric ulcer group, age, race and NSAID use was not found to influence H. pylori prevalence. The prevalence of H. pylori among our peptic ulcer patients is slightly lower compared to overseas studies but the local trend,when compared indirectly to another previous local study did not appear to have changed much. Reasons for the lower H. pylori infection rate in comparison to overseas studies are discussed. The lower H. pylori prevalence among female gastric ulcer patients may be due to the higher prevalence of NSAID use.


Injection Reactions after Administration of Sustained-release Meloxicam to BALB/cJ, C57BL/6J, and Crl:CD1(ICR) Mice.
The sustained-release formulation of meloxicam (MSR) is a compounded NSAID that may provide pain relief for as long as 72 h after administration. MSR injection-site skin reactions have occurred in several species but have not previously been observed in mice. We investigated the development and progression of localized skin reactions after a single injection of MSR in Crl:CD1(ICR), C57BL/6J, and BALB/cJ mice. Each mouse received a subcutaneous injection of MSR (n = 60), standard-formulation meloxicam (MEL; n = 24) or saline (control; SC; n = 24) and was scored daily according to a 5-point system for erythema and mass characteristics. Mice were euthanized at either 7 or 14 d after injection and underwent postmortem analysis. MSR-treated mice had more erythematous and mass reactions than did MEL and SC mice. Mass lesions developed in 49 MSR mice (82%; 95% CI, 70% to 90%), 5 MEL animals (21%; 95% CI, 7% to 42%), and 1 SC mouse. MSR-treated BALB/cJ developed erythematous lesions less frequently than similarly treated Crl:CD1(ICR) or C57BL/6J. Lesions often were ventrolateral to the injection site. The median times to the appearance of mass and erythematous lesions were 2 d and 3 d, respectively. Histologically, the erythematous and mass reactions correlated with necrotizing to pyogranulomatous injection-site panniculitis. Inflammation severity scores at 7 and 14 d after injection were greater in the MSR-treated group than the other 2 groups. No strain- or sex-associated differences emerged except that inflammation severity scores at day 14 were higher in Crl:CD1(ICR) females than males. The character of the inflammatory response in MSR-treated mice did not differ between 7 and 14 d after injection, indicating that MSR-induced inflammation is slow to resolve. The ventral migration and delayed onset of MSR injection-site reactions could result in their being attributed to another cause or not being identified. Researchers and clinicians should be aware of the potential for slowly resolving injection-site reactions with MSR.


Renal safety of tenofovir disoproxil fumarate and entecavir with hepatitis B immunoglobulin in liver transplant patients.
Potent nucleos(t)ide analogues and hepatitis B immunoglobulin combinations are recommended after liver transplantation to prevent the recurrence of hepatitis B virus (HBV). Despite its proven efficacy, the renal safety of tenofovir disoproxil fumarate (TDF) has not been well established in liver transplant recipients. We aimed to assess the impacts of TDF and entecavir (ETV) on tubular and glomerular functions. We analysed 206 liver transplant patients treated with TDF (n = 102) or ETV (n = 104) plus hepatitis B immunoglobulin. Serum creatinine, phosphate and uric acid levels were measured. Proximal tubular dysfunction was defined as the presence of hypophosphatemia (<2 mg/dL) and hypouricemia (<2 mg/dL). Glomerular dysfunction was defined as an estimated glomerular filtration rate (eGFR) of <60 mL/min/1.73 m2 accompanied by a ≥25% eGFR decline from baseline. During a median follow-up of 42.5 months, 48 patients developed proximal tubular dysfunction (30.4% and 16.3% in the TDF and ETV groups; P = .017). Serum levels of phosphate and uric acid were significantly lower in the TDF group post-LT. TDF (OR, 2.34; 95% CI, 1.16-4.69; P = .017) and low body mass index (OR, 2.11; 95% CI, 1.06-4.21; P = .034) were independent risk factors for proximal tubular dysfunction. The prevalence of glomerular dysfunction was not significantly different between the two groups (TDF 51.0% and ETV 54.8%; P = .582). TDF significantly increased the risk of proximal tubular dysfunction. Although the effect of TDF on glomerular function was comparable to that of ETV, glomerular dysfunction was common after liver transplant.


Factors contributing to underuse of radiation among younger women with breast cancer.
Evidence-based literature has confirmed the effectiveness of radiation therapy (RT) after breast-conserving surgery (BCS), especially for young women. However, women with young children may be less likely to be compliant. This study explores factors associated with noncompliance of RT among insured young patients. Using the MarketScan Database, we identified the study cohort as women aged 20 to 64 years who had a BCS between January 1, 2004, and December 31, 2009, and had continuous enrollment 12 months before and after the date of BCS. Patients who had any radiation claims within a year of BCS were considered compliant. Adjusted odds of compliance were estimated from logistic regressions for the full sample and age-stratified subgroups. Sensitivity analyses were performed to evaluate the robustness of study findings. All statistical tests were two-sided. Eighteen thousand one hundred twenty of 21 008 (86.25%) nonmetastatic BCS patients received RT. Among patients aged 20 to 64 years, those with children aged 7 to 12 years, those with children aged 13 to 17 years, and those with no children or children aged 18 years or older were more likely to receive RT than patients with at least one child aged less than 7 years (7-12 years: odds ratio (OR) = 1.32, 95% confidence interval (CI) = 1.05 to 1.66, P = .02; 13-17 years: OR = 1.41, 95% CI = 1.13 to 1.75, P = .002; no children or ≥18 years: OR = 1.38, 95% CI = 1.13 to 1.68, P = .001). Stratified analyses showed that the above association was primarily driven by women in the youngest age group (aged 20-50). Other important factors included breast cancer quality of care measures, enrollment in health maintenance organizations or capitated preferred provider organizations, travelled to a Census division outside their residence for BCS, and whether patients were primary holders of the insurance policy. Competing demands from child care can constitute a barrier to complete guideline-concordant breast cancer therapy. Younger patients may be confronted by unique challenges that warrant more attention in future research.


Staff smoking and other barriers to nicotine dependence intervention in addiction treatment settings: a review.
The aims of this review were to assess smoking prevalence among drug abuse treatment staff and summarize the range of barriers to provision of nicotine dependence intervention to clients receiving addictions treatment. A systematic literature search was conducted to identify publications reporting on workforce smoking prevalence, attitudes toward smoking, and perceived barriers to providing smoking cessation treatment in drug abuse treatment settings. Twenty papers met study inclusion criteria. Staff smoking prevalence estimates in the literature ranged from 14% to 40%. The most frequently reported barriers to providing nicotine dependence intervention in addiction treatment settings were lack of staff knowledge or training in this area, that smoking cessation concurrent with other drug or alcohol treatment may create a risk to sobriety, and staff are themselves smokers. Staff smoking is not uniformly elevated in the drug abuse treatment workforce. Smoking prevalence may be lower where staff are more educated or professionally trained, and may be higher in community-based drug treatment programs. Barriers to treating nicotine dependence may be addressed through staff training, policy development, and by supporting staff to quit smoking. State departments of alcohol and drug programs, and national and professional organizations, can also support treatment of nicotine dependence in drug abuse treatment settings.


Bone marrow mesenchymal stromal cells to treat complications following allogeneic stem cell transplantation.
Allogeneic hematopoietic stem cell transplantation (HSCT) is a technologically complicated procedure that represents the only cure for many hematologic malignancies. However, HSCT is often complicated by life-threatening toxicities related to the chemo-radiation conditioning regimen, poor engraftment of donor HSCs, the hyperinflammatory syndrome of graft-versus-host disease (GVHD), infection risks from immunosuppression, and end-organ damage. Bone marrow stromal cells (MSCs), also known as "mesenchymal stromal cells," not only play a nurturing role in the hematopoietic microenvironment but also can differentiate into other cell types of mesenchymal origin. MSCs are poorly immunogenic, and they can modulate immunological responses through interactions with a wide range of innate and adaptive immune cells to reduce inflammation. They are easily expanded ex vivo and after infusion, home to sites of injury and inflammation to promote tissue repair. Despite promising early trial results in HSCT with significant responses that have translated into survival benefits, there have been significant barriers to successful commercialization as an off-the-shelf therapy. Current efforts with MSCs in the HSCT setting are geared toward determining the factors determining potency, understanding the precise mechanisms of action in human HSCT, knowing their kinetics and fate, optimizing dose and schedule, incorporating biomarkers as response surrogates, addressing concerns about safety, optimizing clinical trial design, and negotiating the uncharted regulatory landscape for licensable cellular therapy.


Characterization and functional biology of the soybean aleurone layer.
Soybean is a globally important oil seed crop. Both the high protein and oil content of soybean seeds make this crop a lucrative commodity. As in higher eukaryotic species with available genomes, the functional annotation of most of soybean's genes still remains to be investigated. A major hurdle in the functional genomics of soybean is a rapid method to test gene constructs before embarking on stable transformation experiments. In this paper we describe the morphology and composition of the persistent single-cell aleurone layer that derives from the endosperm of developing soybean seeds. Its composition compared to cotyledonary tissue indicates the aleurone layer plays a role in both abiotic and biotic stress. The potential utility as the aleurone layer as a transient expression system in soybean was shown. As a near transparent single-cell layer it can be used as a transient expression system to study transgene expression and inter- and intra-cellular targeting as it is amenable to microscopic techniques. The transparent single cell aleurone layer was shown to be compositionally comparable to cotyledonary tissue in soybean with an enrichment in oxidative response proteins and shown to be a potential transient expression platform.


Development of a Decellularized Porcine Esophageal Matrix for Potential Applications in Cancer Modeling.
Many decellularized extracellular matrix-derived whole organs have been widely used in studies of tissue engineering and cancer models. However, decellularizing porcine esophagus to obtain decellularized esophageal matrix (DEM) for potential biomedical applications has not been widely investigated. In this study a modified decellularization protocol was employed to prepare a porcine esophageal DEM for the study of cancer cell growth. The cellular removal and retention of matrix components in the porcine DEM were fully characterized. The microstructure of the DEM was observed using scanning electronic microscopy. Human esophageal squamous cell carcinoma (ESCC) and human primary esophageal fibroblast cells (FBCs) were seeded in the DEM to observe their growth. Results show that the decellularization process did not cause significant loss of mechanical properties and that blood ducts and lymphatic vessels in the submucosa layer were also preserved. ESCC and FBCs grew on the DEM well and the matrix did not show any toxicity to cells. When FBS and ESCC were cocultured on the matrix, they secreted more periostin, a protein that supports cell adhesion on matrix. This study shows that the modified decellularization protocol can effectively remove the cell materials and maintain the microstructure of the porcine esophageal matrix, which has the potential application of studying cell growth and migration for esophageal cancer models.


Why does the yellow-eyed Ensatina have yellow eyes? Batesian mimicry of Pacific newts (genus Taricha) by the salamander Ensatina eschscholtzii xanthoptica.
Color patterns commonly vary geographically within species, but it is rare that such variation corresponds with divergent antipredator strategies. The polymorphic salamander Ensatina eschscholtzii, however, may represent such a case. In this species, most subspecies are cryptically colored, whereas E. e. xanthoptica, the Yellow eyed ensatina, is hypothesized to be an aposematic mimic of highly toxic Pacific newts (genus Taricha). To test the mimicry hypothesis, we conducted feeding trials using Western Scrub-Jays, Aphelocoma californica. In every feeding trial, we found that jays, following presentation with the presumed model (T. torosa), were more hesitant to contact the presumed mimic (E. e. xanthoptica) than a control subspecies lacking the postulated aposematic colors (E. e. oregonensis). The median time to contact was 315 sec for the mimic and 52 sec for the control. These results support the mimicry hypothesis, and we suggest that E. e. xanthoptica is likely a Batesian mimic, rather a Müllerian or quasi-Batesian mimic, of Pacific newts.


Michael addition of thiols to alpha-enones in ionic liquids with and without organocatalysts.
Seventeen organocatalyts were tested for their ability to catalyst the addition of thiophenols to chalcones in [bmim]PF6. The products were isolated in high yield after a short reaction time, but no stereoselectivity was observed. The reactions also proceeded (without any stereoselectivity) in four other ionic liquids. In contrast, 16% and 26% ee were observed when L-proline and cinchonine, respectively, were used as the catalysts in CH2Cl2. Addition of thiophenols is also catalysed by HCl, as well as D-mandelic and L-tartaric acids. Addition of thiophenols to chalcones also occurred in neat ionic liquids, without any additional catalyst, but the rate of the reaction depended considerably on the structure of ionic liquid. The scope of the non-catalysed reaction in ionic liquids was tested by the reactions of 5 different thiols and 3 different alpha-enones.


Similar exemplar pooling processes underlie the learning of facial identity and handwriting style: Evidence from typical observers and individuals with Autism.
Considerable research has addressed whether the cognitive and neural representations recruited by faces are similar to those engaged by other types of visual stimuli. For example, research has examined the extent to which objects of expertise recruit holistic representation and engage the fusiform face area. Little is known, however, about the domain-specificity of the exemplar pooling processes thought to underlie the acquisition of familiarity with particular facial identities. In the present study we sought to compare observers' ability to learn facial identities and handwriting styles from exposure to multiple exemplars. Crucially, while handwritten words and faces differ considerably in their topographic form, both learning tasks share a common exemplar pooling component. In our first experiment, we find that typical observers' ability to learn facial identities and handwriting styles from exposure to multiple exemplars correlates closely. In our second experiment, we show that observers with Autism Spectrum Disorder (ASD) are impaired at both learning tasks. Our findings suggest that similar exemplar pooling processes are recruited when learning facial identities and handwriting styles. Models of exemplar pooling originally developed to explain face learning, may therefore offer valuable insights into exemplar pooling across a range of domains, extending beyond faces. Aberrant exemplar pooling, possibly resulting from structural differences in the inferior longitudinal fasciculus, may underlie difficulties recognising familiar faces often experienced by individuals with ASD, and leave observers overly reliant on local details present in particular exemplars.


The role of the JAK/STAT3 signaling pathway in acquired corneal diseases.
Acquired corneal diseases such as dry eye disease (DED), keratitis and corneal alkali burns are significant contributors to vision impairment worldwide, and more effective and innovative therapies are urgently needed. The Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) signaling pathway plays an indispensable role in cell metabolism, inflammation and the immune response. Studies have shown that regulators of this pathway are extensively expressed in the cornea, inducing significant activation of JAK/STAT3 signaling in specific acquired corneal diseases. The activation of JAK/STAT3 signaling contributes to various pathophysiological processes in the cornea, including inflammation, neovascularization, fibrosis, and wound healing. In the context of DED, the hypertonic environment activates JAK/STAT3 signaling to stimulate corneal inflammation. Inflammation and injury progression in infectious keratitis can also be modulated by JAK/STAT3 signaling. Furthermore, JAK/STAT3 signaling is involved in every stage of corneal repair after alkali burns, including acute inflammation, angiogenesis and fibrosis. Treatments modulating JAK/STAT3 signaling have shown promising results in attenuating corneal damage, indicating its potential as a novel therapeutic target. Thus, this review emphasizes the multiple roles of the JAK/STAT3 signaling pathway in common acquired corneal disorders and summarizes the current achievements of JAK/STAT3-targeting therapy to provide new insights into future applications.


Effect of liquid cow manure on andisol properties and atrazine adsorption.
Application of animal manure amendments to agricultural soils is a common practice to improve soil fertility through the addition of essential plant nutrients. This practice may increase the potential for atrazine (2-chloro-4-ethylamino-6-isopropylamino-1, 3, 5-triazine) leaching due to competition for adsorption sites between the pesticide and dissolved organic carbon (DOC) added through manure. We evaluated the influence of liquid cow manure (LCM) application on soil properties, atrazine adsorption, and the physicochemical controlling mechanisms in an Andisol. The LCM was applied at rates equivalent to 0, 100,000, 200,000, and 300,000 L ha(-1), resulting in treatments S-0, S-100, S-200, and S-300, respectively. The LCM application increased DOC and pH of the soils immediately on addition, but pH returned to S-0 values 30 d after application. The LCM application did not modify atrazine adsorption with the two lowest application rates (S-100 and S-200), but atrazine adsorption was decreased in S-300 (K(f) = 0.96) compared with the control (S-0) (K(f) = 1.19), possibly due to the competitive adsorption of DOC with the pesticide. The Fourier-transformed infrared analysis showed that LCM increased aliphaticity and presence of N-containing groups and polysaccharide-like groups in amended soils; however, these properties did not modify the atrazine interaction in the studied amended soils. Interestingly the addition of DOC to soil at the high application rate (S-300) reduced atrazine adsorption in this rich OM Andisol despite the LCM not raising the concentration of stable organic matter. The application of high rates of liquid manure containing DOC incurs an increased risk of pesticide leaching.


Quantitative determination of optical trapping strength and viscoelastic moduli inside living cells.
With the success of in vitro single-molecule force measurements obtained in recent years, the next step is to perform quantitative force measurements inside a living cell. Optical traps have proven excellent tools for manipulation, also in vivo, where they can be essentially non-invasive under correct wavelength and exposure conditions. It is a pre-requisite for in vivo quantitative force measurements that a precise and reliable force calibration of the tweezers is performed. There are well-established calibration protocols in purely viscous environments; however, as the cellular cytoplasm is viscoelastic, it would be incorrect to use a calibration procedure relying on a viscous environment. Here we demonstrate a method to perform a correct force calibration inside a living cell. This method (theoretically proposed in Fischer and Berg-Sørensen (2007 J. Opt. A: Pure Appl. Opt. 9 S239)) takes into account the viscoelastic properties of the cytoplasm and relies on a combination of active and passive recordings of the motion of the cytoplasmic object of interest. The calibration procedure allows us to extract absolute values for the viscoelastic moduli of the living cell cytoplasm as well as the force constant describing the optical trap, thus paving the way for quantitative force measurements inside the living cell. Here, we determine both the spring constant of the optical trap and the elastic contribution from the cytoplasm, influencing the motion of naturally occurring tracer particles. The viscoelastic moduli that we find are of the same order of magnitude as moduli found in other cell types by alternative methods.


Physiological effects of nanoparticles on fish: a comparison of nanometals versus metal ions.
The use of nanoscale materials is growing exponentially, but there are also concerns about the environmental hazard to aquatic biota. Metal-containing engineered nanoparticles (NPs) are an important group of these new materials, and are often made of one metal (e.g., Cu-NPs and Ag-NPs), metal oxides (e.g., ZnO and TiO(2) NPs), or composite of several metals. The physiological effects and toxicity of trace metals in the traditional dissolved form are relatively well known and the overall aim of this review was to use our existing conceptual framework of metal toxicity in fish to compare and contrast the effects of nanometals. Conceptually, there are some fundamental differences that relate to bioavailability and uptake. The chemistry and behaviour of nanometals involves dynamic aspects of aggregation theory, rather than the equilibrium models traditionally used for free metal ions. Some NPs, such as Cu-NPs, may also release free metal ions from the surface of the particle. Biological uptake of NPs is not likely via ion transporters, but endocytosis is a possible uptake mechanism. The body distribution, metabolism, and excretion of nanometals is poorly understood and hampered by a lack of methods for measuring NPs in tissues. Although data sets are still limited, emerging studies on the acute toxicity of nanometals have so far shown that these materials can be lethal to fish in the mg-μgl(-1) range, depending on the type of material. Evidence suggests that some nanometals can be more acutely toxic to some fish than dissolved forms. For example, juvenile zebrafish have a 48-h LC(50) of about 0.71 and 1.78mgl(-1) for nano- and dissolved forms of Cu respectively. The acute toxicity of metal NPs is not always explained, or only partly explained, by the presence of free metal ions; suggesting that other novel mechanisms may be involved in bioavailability. Evidence suggests that nanometals can cause a range of sublethal effects in fish including respiratory toxicity, disturbances to trace elements in tissues, inhibition of Na(+)K(+)-ATPase, and oxidative stress. Organ pathologies from nanometals can be found in a range of organs including the gill, liver, intestine, and brain. These sublethal effects suggest some common features in the sublethal responses to nanometals compared to metal salts. Effects on early life stages of fish are also emerging, with reports of nanometals crossing the chorion (e.g., Ag-NPs), and suggestions that the nano-forms of some metals (Cu-NPs and ZnO NPs) may be more toxic to embryos or juveniles, than the equivalent metal salt. It remains possible that nanometals could interfere with, and/or stimulate stress responses in fish; but data has yet to be collected on this aspect. We conclude that nanometals do have adverse physiological effects on fish, and the hazard for some metal NPs will be different to the traditional dissolved forms of metals.


Heritability and correlations among learning and inhibitory control traits.
To understand the evolution of cognitive abilities, we need to understand both how selection acts upon them and their genetic (co)variance structure. Recent work suggests that there are fitness consequences for free-living individuals with particular cognitive abilities. However, our current understanding of the heritability of these abilities is restricted to domesticated species subjected to artificial selection. We investigated genetic variance for, and genetic correlations among four cognitive abilities: inhibitory control, visual and spatial discrimination, and spatial ability, measured on >450 pheasants, Phasianus colchicus, over four generations. Pheasants were reared in captivity but bred from adults that lived in the wild and hence, were subject to selection on survival. Pheasant chicks are precocial and were reared without parents, enabling us to standardize environmental and parental care effects. We constructed a pedigree based on 15 microsatellite loci and implemented animal models to estimate heritability. We found moderate heritabilities for discrimination learning and inhibitory control (h2 = 0.17-0.23) but heritability for spatial ability was low (h2 = 0.09). Genetic correlations among-traits were largely positive but characterized by high uncertainty and were not statistically significant. Principle component analysis of the genetic correlation matrix estimate revealed a leading component that explained 69% of the variation, broadly in line with expectations under a general intelligence model of cognition. However, this pattern was not apparent in the phenotypic correlation structure which was more consistent with a modular view of animal cognition. Our findings highlight that the expression of cognitive traits is influenced by environmental factors which masks the underlying genetic structure.


A discriminative framework for detecting remote protein homologies.
A new method for detecting remote protein homologies is introduced and shown to perform well in classifying protein domains by SCOP superfamily. The method is a variant of support vector machines using a new kernel function. The kernel function is derived from a generative statistical model for a protein family, in this case a hidden Markov model. This general approach of combining generative models like HMMs with discriminative methods such as support vector machines may have applications in other areas of biosequence analysis as well.


How patients' self-disclosure about sickle cell pain episodes to significant others relates to living with sickle cell disease.
This cross-sectional study examines to whom and how fully sickle cell disease (SCD) patients talk to others about sickle cell pain, how helpful it is to talk with others about these pain episodes, and the association between talking to others about sickle cell pain episodes and patients' psychological adjustment and coping strategies in managing the disease. A convenience sample of 73 African American patients with SCD (30 men and 43 women), were recruited from two SCD clinics at the time of routine medical visits. Most participants had been diagnosed with hemoglobin SS, and they reported an average number of 8.61 pain episodes in the previous 12 months. Participants were asked to whom, how fully, and how helpful it was to talk to significant others about SCD pain episodes experienced in the last 12 months. Patients also completed measures of their psychological adjustment as well as how they would manage a future sickle cell pain episode. Self-report ratings were made on Likert-type scales. Based on paired samples t-tests, participants talked significantly more fully about their thoughts and feelings concerning pain episodes to God and to their primary medical providers than to either their parents, siblings, or an intimate partner/close friend. Bivariate correlations indicated that amount and helpfulness of talking about pain episodes to God and to parents were significantly associated with better psychological adjustment on selected measures. Also, bivariate correlations indicated that helpfulness in talking with siblings, intimate partner/close friend, and primary medical providers was positively related with willingness to go to a physician in the event of a future pain episode. The results document to whom and how helpful it is to talk with others about SCD pain episodes and how SCD disclosure is related to strategies for managing this disease.


Chiari malformation presenting as a focal motor deficit. Report of two cases.
Chiari malformations may present with a wide range of symptoms and signs. Nevertheless, focal foot weakness as a presentation of a Chiari malformation has not been described in the pediatric neurosurgical literature. Two children with Chiari malformations and holocord syringomyelia presented with manifestations of a supposed isolated lumbar radiculopathy. Neurological deficits completely resolved after decompressive suboccipital craniectomy and cervical laminectomy. These cases emphasize the importance of imaging the entire craniospinal axis and avoidance of therapeutic intervention specifically aimed at a radiculopathic process when initial imaging fails to show a structural abnormality at the spinal level of deficit. The possible pathophysiological origins for this unusual presentation are discussed. Based on the experience gained with these patients, recommendations are made regarding the diagnostic workup and management of this entity in children presenting with focal deficits that are not supported by imaging of the affected root levels. Chiari malformations may rarely masquerade as lower motor and sensory deficits, and appropriate treatment may result in excellent recovery of function.


Vismodegib (ERIVEDGE°) In basal cell carcinoma: too many unknowns.
Basal cell carcinomas are the most common skin cancers. They are usually localised and carry a good prognosis. There is no standard treatment for the rare patients with metastatic basal cell carcinoma or very extensive basal cell carcinoma for whom surgery or radiotherapy is inappropriate. Vismodegib, a cytotoxic drug, is claimed to prevent tumour growth by inhibiting a pathway involved in tissue repair and embryogenesis. It has been authorised in the European Union for patients with metastatic or locally advanced and extensive basal cell carcinoma. Clinical evaluation of vismodegib is based on a non-comparative clinical trial involving 104 patients, providing only weak evidence. Twenty-one months after the start of the trial, 7 patients with metastases (21%) and 6 patients with advanced basal cell carcinoma (10%) had died. Given the lack of a placebo group, there is no way of knowing whether vismodegib had any effect, positive or negative, on survival. There were no complete responses among patients with metastases, but about one-third of them had partial responses. Among the 63 patients with locally advanced basal cell carcinoma, there were 14 complete responses and 16 partial responses. The recurrence rate in patients with complete responses was not reported. Similar results were reported in two other uncontrolled trials available in mid-2014. Vismodegib has frequent and sometimes serious adverse effects, including muscle spasms, fatigue and severe hyponatraemia. Cases of severe weight loss, alopecia, ocular disorders, other cancers (including squamous cell carcinoma) and anaemia have also been reported. More data are needed on possible hepatic and cardiovascular adverse effects. A potent teratogenic effect was seen in experimental animals. As vismodegib enters semen, contraception is mandatory for both men (condoms) and women. In practice, vismodegib has frequent and varied adverse effects, some of which are serious, while its benefits are poorly documented. Vismodegib should only be proposed to patients in whom basal cell cancer markedly undermines quality of life, and only in the context of clinical research.


Src inhibitor reduces permeability without disturbing vascularization and prevents bone destruction in steroid-associated osteonecrotic lesions in rabbits.
To examine the therapeutic effect of Src inhibitor on the VEGF mediating vascular hyperpermeability and bone destruction within steroid-associated osteonecrotic lesions in rabbits. Rabbits with high risk for progress to destructive repair in steroid-associated osteonecrosis were selected according to our published protocol. The selected rabbits were systemically administrated with either Anti-VEGF antibody (Anti-VEGF Group) or Src inhibitor (Src-Inhibition Group) or VEGF (VEGF-Supplement Group) or a combination of VEGF and Src inhibitor (Supplement &Inhibition Group) or control vehicle (Control Group) for 4 weeks. At 0, 2 and 4 weeks after administration, in vivo dynamic MRI, micro-CT based-angiography, histomorphometry and immunoblotting were employed to evaluate the vascular and skeletal events in different groups. The incidence of the destructive repair in the Anti-VEGF Group, Src-Inhibition Group and Supplement &Inhibition Group was all significantly lower than that in the Control Group. The angiogenesis was promoted in VEGF-Supplement Group, Src-Inhibition Group and Supplement &Inhibition Group, while the hyperpermeability was inhibited in Anti-VEGF Group, Src-Inhibition Group and Supplement &Inhibition Group. The trabecular structure was improved in Src-Inhibition Group and Supplement &Inhibition Group. Src inhibitor could reduce permeability without disturbing vascularization and prevent destructive repair in steroid-associated osteonecrosis.


Associations of insomnia symptoms with sociodemographic, clinical, and lifestyle factors in persons with HF: Health and retirement study.
Insomnia symptoms are very common in persons with heart failure (HF). However, many of the correlates and predictors of insomnia symptoms in this population remain unclear. The purpose of this study is to investigate the associations of sociodemographic, clinical, and lifestyle factors with insomnia symptoms in persons with HF. A theoretical framework was adapted from the neurocognitive model of chronic insomnia to guide the study. Data from the health and retirement study were used for the analysis. Parametric and nonparametric bivariate and multivariate analyses were conducted to investigate these associations. Age, depressive symptoms, comorbidity, dyspnea, pain, and smoking had significant bivariate associations with all insomnia symptoms. Race, Hispanic ethnicity, marital status, household income, poverty, and physical activity were associated with difficulty initiating sleep (DIS) and early morning awakening (EMA). Female sex, education, and alcohol consumption had a significant bivariate association with DIS. Sleep-disordered breathing and body mass index were significantly associated with EMA. Multivariate analysis suggested that depressive symptoms, comorbidity, dyspnea, and pain had independent associations with each insomnia symptom. Age explained DIS and difficulty maintaining sleep, and significant interaction effects between age and physical activity on DIS and EMA were revealed. Results suggest that insomnia symptoms are associated with several sociodemographic, clinical, and lifestyle factors. Age below 70 years, depressive symptoms, comorbidity, dyspnea, and pain might be considered as a phenotype to identify persons with HF who are at increased risk for insomnia symptoms.


Evidence for distinct calcium channel agonist and antagonist binding sites in intact cultured embryonic chick ventricular cells.
To determine whether calcium channel agonists and antagonists bind to distinct pharmacologically active sites, the binding of dihydropyridine calcium channel agonists and antagonists was related to calcium flux and contractile state in primary monolayer cultures of spontaneously contracting chick embryo ventricular cells. Equilibrium binding studies using the antagonist (+)-[3H]PN200-110 demonstrated equilibrium binding to intact, beating cells consistent with a single class of binding sites (KD, 1.1 nM; Bmax, 40 fmol/mg protein). Membrane depolarization of the intact cells by incubation in 30 mM potassium caused a 91% increase in the apparent number of (+)-PN200-110 binding sites (Bmax 76 fmol/mg protein), but no significant change in the KD (1.2 nM). The (+)-PN200-110 produced a concentration-dependent decrease in calcium uptake (IC50 2.2 nM) and contractile amplitude (IC50 5.6 nM). The calcium channel agonist, (+/-)-[3H]BAY k 8644, bound to two distinct binding sites with high affinity (KD 1.0 nM) and low affinity (KD 1.9 microM). The (+/-)-BAY k 8644 produced biphasic modulation of calcium flux and contractile state. At concentrations of 100 nM or less, (+/-)-BAY k 8644 increased calcium flux and contractile amplitude, consistent with drug interaction with the high affinity agonist site. However, at higher concentrations, the stimulatory effect of (+/-)-BAY k 8644 on calcium flux and contractile amplitude was abolished, a finding that is consistent with drug interaction with the low affinity antagonist site.(ABSTRACT TRUNCATED AT 250 WORDS)


Is there any benefit of preservation of the spleen in distal pancreatectomy?
For a pancreatic body or tail tumor, distal pancreatectomy with splenectomy (DPS) is a standard operation. Spleen-preserving distal pancreatectomy (SPDP) was introduced in order to preserve the organ and thus provide the patient with a better quality of life. Clinical data were compared between 38 Japanese patients with DPS and 9 with SPDP for benign tumors or tumor-like lesions at the body or tail of the pancreas at preoperative, early postoperative (< 3 months after operation), and late postoperative periods (>6 months after operation). The preoperative findings were not different between the two groups except for the significantly higher serum amylase levels in the SPDP group. Operation time, operative blood loss, and length of postoperative hospital stay were not different between the two groups. Pancreatic fistula occurred in 3 (8%) of the 38 patients in the DPS group and in 1 (11%) of the 9 patients in the SPDP group, abdominal abscess in 5 (13%) of the 38 patients in the DPS group and none (0%) in the 9 patients in the SPDP group. At short-term, clinical findings were not different between the two groups except for a significantly greater platelet count in the DPS group than in the SPDP group (46.8 x 10(4)/microl versus 29.6 x 10(4)/microl, P = 0.0081). At long-term after the operation, clinical findings, including the platelet count, were not different between the two groups. Computed tomography revealed a pseudocyst in 9 (53%) of 17 patients examined in the DPS group and in 3 (75%) of 4 patients examined in the SPDP group at short-term after operation. All patients with pseudocysts were asymptomatic. Two asymptomatic patients (one in the DPS group and one in the SPDP group) first developed a pseudocyst at long-term after the operation. The alteration of glucose tolerance was similar between the two groups. Postoperative pancreatic exocrine function (the N-benzol-L-tyrosyl-p-aminobenzoic acid test) was not different between the two groups. These data suggest that SPDP with preservation of the splenic vessels can be satisfactorily performed without elongating operative time and postoperative hospital stay or increasing risk of postoperative complications, with the exception of increased platelet count in the DPS group at short-term after the operation. Thus, SPDP is worth considering as one of the options for the treatment of benign lesions of the body or tail of the pancreas.


What Is in the Literature.
What is in the Literature focuses on peripheral neuropathies with new and practical information related to the diagnosis, treatment, and management. Diagnostic and treatment guidelines are available for chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) but not all clinicians follow them resulting in erroneous diagnoses and prolonged treatment. Secondary axonal loss in CIDP causes increased connective tissue in muscle. Antibodies to proteins at the node of Ranvier are found in a small percentage of patients with CIDP. The differential diagnosis for CIDP-like neuropathies includes amyloid neuropathy and POEMS (polyneuropathy, organomegaly, endocrinopathy, M-protein, skin changes) and amyloidosis. Upper limits for cerebral spinal fluid protein are 0.45 g/L and cell count <10/µL, but both may be too low. Hyperactive reflexes may occur in Guillain-Barré syndrome and should not exclude the diagnosis. In severely affected Guillain-Barré syndrome patients, a second dose of intravenous immune globulin within 4 weeks of onset is not likely to be effective.


Invisibility, safety and psycho-social distress among same-sex attracted women in rural South Australia.
Available work from North America indicates that same-sex attracted (SSA) individuals enjoy aspects of rural life but nonetheless report encountering homophobia and experiencing isolation from SSA networks. The experience of prejudice and social isolation are often associated with psycho-social distress among the general population of same-sex attracted individuals. Little is known of how SSA women experience life in rural areas of Australia and how this influences their psycho-social wellbeing. This was a small-scale qualitative study using guided interviews to explore the experience of SSA women living in rural areas of South Australia. Seven women identifying as same-sex attracted were interviewed. In addition, a woman who provides a counseling and support service for same-sex attracted women was also interviewed. All interviews were audiotaped and transcribed verbatim and were then analysed for emergent themes. Summaries of the interviews, based on the emergent themes, were sent to all interviewees so that they could verify or challenge the validity of the emergent themes, as well as to allow them to remove any information they felt might identify them. Most women had felt 'different' while growing up; almost unanimously describing themselves as having been 'tomboys'. However, the lack of visible SSA role models in rural areas, together with a lack of SSA social networks, did not allow some of the women to identify and name their same-sex attraction. For many of the women in this study, it was visits to the state capital, where they had the opportunity to meet other SSA women, which precipitated them identifying themselves as same-sex attracted. In light of this new knowledge, some women denied their same-sex attraction and entered into heterosexual relationships, often entailing marriage. Other women entered same-sex relationships but tried to keep them invisible within their communities. Rural communities are frequently close-knit environments, where 'everybody knows everybody'. In such settings, much of the conduct of daily life is visible to other community members and may be closely scrutinized. In such locations, women initially try to keep their same sex attraction invisible. Women rarely comment on the threat of violence that underpins the attempt to keep same-sex identity or relationships secret. Nevertheless, even while it remains a largely unarticulated factor, it was a powerful one fueling the maintenance of their invisibility. Fear of violence ranges from the threat of overt physical violence to more commonly voiced concerns over 'mundane' harassment. Fear of social violence, in the form of rejection and ostracism, is the most frequently nominated factor motivating women to keep same-sex attraction invisible in rural settings. While keeping their same-sex identity and relationships invisible offers women some safety from physical and social violence, it may pose dangers for their psycho-social well-being. Trying to keep SSA relationships invisible within communities involves women censoring their behaviour while in public. Invisibility also gives rise to isolation, since women do not have the opportunity to form same-sex attracted social networks. Furthermore, some women experience great psycho-social distress as their awareness of their same-sex attraction becomes increasingly incongruent with the established pattern of their lives. Women find it very difficult to seek appropriate forms of help if they do experience psycho-social problems. Concerns over confidentiality means that women are extremely reluctant to seek help and support locally. Distance is a powerful mediating factor in both women's experience of distress and their capacity to seek help to ameliorate it. For women who live in closer proximity to the state capital, the capacity to meet other SSA women and engage in same-sex attracted social activities helps ease their sense of difference and isolation. Likewise, should they wish to utilize counseling or support services, it is relatively easy for them to do so. However, women living at greater distances from a capital city experience greater isolation and loneliness. Moreover, their ability to confidentially access appropriate services is markedly curtailed. Same-sex attracted women living in rural areas need social contact with other SSA women, where their safety and confidentiality can be assured. They also need access to safe, confidential and appropriate services. Telephone and internet services are one way of anonymously and confidentially giving women access to information and support. Peer support networks, internet and telephone networks need to be further developed. Ways in which specialist gay and lesbian services and locally based health and human services could collaborate to provide accessible and acceptable services and to help women develop safe and confidential social networks need to be identified and trialled.


Development of mouse models for analysis of human virus infections.
Viruses usually exhibit strict species-specificity as a result of co-evolution with the host. Thus, in mouse models, a great barrier exists for analysis of infections with human-tropic viruses. Mouse models are unlikely to faithfully reproduce the human immune response to viruses or viral compounds and it is difficult to evaluate human therapeutic efficacy with antiviral reagents in mouse models. Humans and mice essentially have different immune systems, which makes it difficult to extrapolate mouse results to humans. In addition, apart from immunological reasons, viruses causing human diseases do not always infect mice because of species tropism. One way to determine tropism would be a virus receptor that is expressed on affected cells. The development of gene-disrupted mice and Tg mice, which express human receptor genes, enables us to analyze several viral infections in mice. Mice are, indeed, susceptible to human viruses when artificially infected in receptor-supplemented mice. Although the mouse cells less efficiently permit viral replication than do human cells, the models for analysis of human viruses have been established in vivo as well as in vitro, and explain viral pathogenesis in the mouse systems. In most systems, however, nucleic acid sensors and type I interferon suppress viral propagation to block the appearance of infectious manifestation. We herein review recent insight into in vivo antiviral responses induced in mouse infection models for typical human viruses.


Quantitative detection method for Roundup Ready soybean in food using duplex real-time PCR MGB chemistry.
Methodologies that enable the detection of genetically modified organisms (GMOs) (authorized and non-authorized) in food and feed strongly influence the potential for adequate updating and implementation of legislation together with labeling requirements. Quantitative polymerase chain reaction (qPCR) systems were designed to boost the sensitivity and specificity on the identification of GMOs in highly degraded DNA samples; however, such testing will become economically difficult to cope with due to increasing numbers of approved genetically modified (GM) lines. Multiplexing approaches are therefore in development to provide cost-efficient solution. Construct-specific primers and probe were developed for quantitative analysis of Roundup Ready soybean (RRS) event glyphosate-tolerant soybean (GTS) 40-3-2. The lectin gene (Le1) was used as a reference gene, and its specificity was verified. RRS- and Le1-specific quantitative real-time PCR (qRTPCR) were optimized in a duplex platform that has been validated with respect to limit of detection (LOD) and limit of quantification (LOQ), as well as accuracy. The analysis of model processed food samples showed that the degradation of DNA has no adverse or little effects on the performance of quantification assay. In this study, a duplex qRTPCR using TaqMan minor groove binder-non-fluorescent quencher (MGB-NFQ) chemistry was developed for specific detection and quantification of RRS event GTS 40-3-2 that can be used for practical monitoring in processed food products.


Usual consumption of plant foods containing phytoestrogens and sex hormone levels in postmenopausal women in Wisconsin.
Consumption of phytoestrogens may reduce hormone-dependent cancer risk through alterations in the actions or metabolism of steroid hormones. Studies in humans of phytoestrogen-hormone interactions have been limited and inconsistent. Relations between the consumption of phytoestrogen-containing foods and serum sex hormones and sex hormone-binding globulin were studied in a population-based sample of postmenopausal women who participated in the Nutritional Factors in Eye Disease Study of the Beaver Dam Eye Study. Information on phytoestrogen-containing foods (broccoli, carrots, cauliflower, chili, dark bread, peas, and dried beans) was collected by interviewer-administered food-frequency questionnaires. Estrone, sex hormone-binding globulin, dehydroepiandrosterone sulfate, and total and free testosterone were measured. Analyses included 246 postmenopausal women not taking hormone replacements. Partial correlations between hormones and intake of phytoestrogen-containing foods were computed, with adjustment for age, body mass index, years since menopause, and total energy intake. Number of standard servings per week of whole-grain products from the dark bread group was inversely associated with total testosterone (r = -0.20, p = 0.002). Although not statistically significant, other hormones displayed similar inverse associations with dark bread consistent with a common metabolic pathway. Although the magnitude of association was small, the data are consistent with the possibility that consumption of some phytoestrogen-containing foods may affect levels of testosterone in postmenopausal women.


A simple pupil-independent method for recording eye movements in rodents using video.
Previous techniques using video to acquire torsional eye-movement recordings in humans have needed to control confounding effects associated with dilation of the pupil. A new pupil-independent, three-dimensional eye-movement recording method was developed to overcome the reliance on tracking the pupil by interpolating between two artificial landmarks placed on the eye. This technique reliably quantified torsional changes in angular eye position at variable amplitudes of horizontal and vertical excursion over at least a 20 degrees range. Application of this system to rodents naturally lacking a clearly defined pupil allows behavioral researchers to obtain torsion in addition to horizontal and vertical changes in angular eye position.


Changes in rat pituitary nuclear and cytoplasmic pro-opiomelanocortin RNAs associated with adrenalectomy and glucocorticoid replacement.
While the transcriptional effects of glucocorticoid hormone manipulation on the pituitary pro-opiomelanocortin (POMC) gene have been documented, it is not yet clear whether glucocorticoids activate additional post-transcriptional mechanisms to regulate POMC gene expression. We have used RNA probes that span exon/intron junctions in sensitive nuclease protection assays in order to examine changes in POMC precursor RNA as well as mature mRNA in nucleus and cytoplasm following both adrenalectomy (ADX) and administration of exogenous glucocorticoids. ADX led to a rapid and sustained 8- to 10-fold increase in the level of POMC primary transcript in the anterior lobe (AL), from 1 to 14 days after ADX. Stimulation of mature POMC mRNA in the nucleus was also rapid, with 7- to 8-fold increases evident by 1 day after ADX. In sharp contrast, the time-dependent accumulation of POMC mRNA in the cytoplasm was slow in comparison, reaching levels approximately 2-fold higher than sham-operated animals by 1 day post-ADX and 12-fold higher by 14 days after ADX. Despite the constant elevated level of nuclear POMC precursor RNA, the rate of accumulation of POMC mRNA in the corticotroph cytoplasm after ADX was not linear, with the greatest increase occurring within the first 1-4 days post-ADX. This led to alterations in the molar ratio of POMC primary transcript: nuclear mRNA: cytoplasmic mRNA in the AL at 1 and 4 days after ADX and showed a relative increase in the proportion of POMC RNA transcripts within the nucleus. Acute administration of dexamethasone to ADX rats resulted in rapid 80-90% inhibition of POMC primary transcript levels in the AL that was maximal by 30 min but with no associated change in mature mRNA. No significant changes in POMC RNA were seen in neurointermediate lobe in any of these studies. These studies suggest that following ADX, time-dependent alterations in nuclear transport of mature POMC mRNA and/or changes in POMC mRNA stability, in addition to changes in gene transcription may account for the overall level of POMC mRNA expressed in the AL. Furthermore, we have illustrated the use of exon/intron probes for accurately quantitating rapid alterations in steady-state levels of nuclear precursor RNA that may reflect transcriptional responses and/or changes in post-transcriptional processing of the primary transcript.


Consanguineous marriage, prepregnancy maternal characteristics and stillbirth risk: a population-based case-control study.
Consanguineous marriage is associated with increased risks for congenital anomalies, low birthweight, and other adverse perinatal outcomes. In this population-based, case-control study we investigated the association between consanguineous marriage (first-cousin marriage) and stillbirth risk, using prospectively collected information from prepregnancy visits. From 2007 to 2009, we identified 283 stillbirths (cases) and 2088 randomly selected live control births through prepregnancy visits in rural Golestan, Iran. The associations between consanguinity and prepregnancy maternal characteristics and stillbirth risk were examined using multivariate logistic regression. The rate of consanguineous marriage was 19.4% among cases and 13.6% among controls. Consanguinity was associated with increased stillbirth risk [odds ratio (OR) 1.53; 95% CI 1.10-2.14]. The association was significantly increased for preterm stillbirth (< 37 gestational weeks) (OR 2.43; 95% CI 1.46-4.04) but not for term stillbirth (≥ 37 weeks) (OR 1.14; 95% CI 0.75-1.74). Low and high maternal age, underweight, obesity, nulliparity, a history of infertility or miscarriage, previous obstetric complications (preeclampsia, preterm delivery, and stillbirth in previous pregnancies) were also associated with increased stillbirth risks. Consanguineous marriage is associated with increased risk of stillbirth, particularly preterm stillbirth. Findings for other maternal risk factors for stillbirth in rural Iran are consistent with previously reported findings from high-income countries.


Domain organization of Salmonella typhimurium formylglycinamide ribonucleotide amidotransferase revealed by X-ray crystallography.
Formylglycinamide ribonucleotide amidotransferase (FGAR-AT) catalyzes the ATP-dependent conversion of formylglycinamide ribonucleotide (FGAR) and glutamine to formylglycinamidine ribonucleotide (FGAM), ADP, P(i), and glutamate in the fourth step of the purine biosynthetic pathway. In eukaryotes and Gram-negative bacteria, FGAR-AT is encoded by the purL gene as a multidomain protein with a molecular mass of about 140 kDa. In Gram-positive bacteria and archaebacteria FGAR-AT is a complex of three proteins: PurS, PurL, and PurQ. We have determined the structure of FGAR-AT (PurL) from Salmonella typhimurium at 1.9 A resolution using X-ray crystallography. PurL is the last remaining enzyme in the purine biosynthetic pathway to have its structure determined. The structure reveals four domains: an N-terminal domain structurally homologous to a PurS dimer, a linker region, an FGAM synthetase domain homologous to an aminoimidazole ribonucleotide synthetase (PurM) dimer, and a triad glutaminase domain. The domains are intricately linked by interdomain interactions and peptide connectors. The fold common to PurM and the central region of PurL represents a superfamily for which HypE, SelD, and ThiL are predicted to be members. A structural ADP molecule was found bound to a site related to the putative active site by pseudo-2-fold symmetry and two sulfate ions were found at the putative active site. These observations and the structural similarities between PurM and StPurL were used to model the substrates FGAR and ATP in the StPurL active site. A glutamylthioester intermediate was found in the glutaminase domain at Cys1135. The N-terminal (PurS-like) domain is hypothesized to form the putative channel through which ammonia passes from the glutaminase domain to the FGAM synthetase domain.


REOX: Evaluation of the Efficacy of Retreatment With an Oxaliplatin-containing Regimen in Metastatic Colorectal Cancer: A Retrospective Single-center Study.
Treatment of metastatic colorectal adenocarcinoma (mCRC) has evolved, and survival is over 30 months in contemporary trials. Nevertheless, there is a paucity of effective regimes after the first or second-line treatment. Thus, reexposure to previously used drugs has become a treatment strategy for some patients. We aimed to evaluate the efficacy of retreatment with an oxaliplatin-containing regimen in mCRC and correlate this with clinicopathologic features. We retrospectively analyzed 83 patients with mCRC who underwent reexposure to oxaliplatin (REOX). REOX was defined as a second trial of an oxaliplatin-containing regimen after a previous failure. Primary endpoint was time to treatment failure (TTF). The median age was 53.5 years, and the female/male ratio was 51.8%/48.2%. The site of the primary tumor was colon (67.5%) and rectal (32.5%). KRAS was mutated in 39.8%. Liver-limited metastasis was found in 19.3% of patients. The main regimen was 5-fluorouracil, levoleucovorin, and oxaliplatin (mFOLFOX6) (84.3%). Bevacizumab and cetuximab were used in 42.2% and 6% of patients, respectively. REOX was used in the third and fourth lines in 48.2% and 25.3% of patients, respectively. The median TTF after REOX was 6.04 months. Overall survival (OS) was 10.04 months. Disease control (complete response + partial response + stable disease) was observed in 56.6%, whereas 42.2% had progressive disease. Partial response + complete response to previous oxaliplatin was predictive of prolonged OS. Patients who attained disease control had better median OS compared with those with progressive disease (14.5 vs. 6.24 months; P < .0001). In the setting of heavily pretreated patients with mCRC, REOX was an effective treatment, with mTTF of 6.04 months in our cohort. Selection of patients with the longest time since previous oxaliplatin can translate in better outcome. Further studies should be conducted to confirm our data.


Evaluation of the French reactive mortality surveillance system supporting decision making.
In France, a mortality syndromic surveillance system was set up with objectives of early detection and reactive evaluation of the impact of expected and unexpected events to support decision makers. This study aims to describe the characteristics of the system and its usefulness for decision makers. Anonymized data from the administrative part of death certificates were daily collected from 3062 computerized city halls and were transmitted to Santé publique France in routine. Coverage of the system was measured as the proportion of deaths registered by the system among the complete number of deaths and analyzed by age, month and region. Deaths were described by gender, age and geographical level using proportion. The excess periods of deaths were described based on the comparison of the weekly observed and expected numbers of deaths between 2012 and 2016. The system recorded 77.5% of the national mortality covering the whole territory. About 81% of deaths were aged 65 years old and more. The surveillance system identified mortality variations mainly during winter and summer, for some concomitant with influenza epidemic or heatwave period, and thus provided information for decision makers. The ability of the system to detect and follow mortality outbreaks in routine in the whole territory has been demonstrated. It is a useful tool to provide early evaluation of the impact of threats on mortality and alert decision makers to adapt control measures. However, the absence of information on medical causes of death may limit the ability to target recommendations.


Reducing the health consequences of smoking--a progress report.
Smoking has been identified as one of the health priority areas to be addressed by the Public Health Service's Objectives for the Nation initiative. Several gains in moving toward the 1990 goals for smoking and health have been recorded. Only 32.6 percent of the U.S. population over 16 years old were smokers in 1980, compared with 41.7 percent in 1965. The proportion of high school seniors who were daily smokers fell from nearly 30 percent in 1977 to 20 percent in 1981. Changes in smoking prevalence were related to critical events, such as the Surgeon General's reports on smoking. A variety of information and education programs aimed at specific groups are being carried out by Federal, State, and local governmental agencies and voluntary health organizations.


Soluble overexpression in Escherichia coli, and purification and characterization of wild-type recombinant tobacco acetolactate synthase.
Acetolactate synthase (ALS) is the first common enzyme in the biosynthesis of L-leucine, L-isoleucine, and L-valine. The wild-type ALS gene from Nicotiana tabacum was cloned into the bacterial expression vector pGEX-2T. The resulting recombinant plasmid pGEX-ALS2 was used to transform Escherichia coli strain XL1-Blue, and the wild-type tobacco ALS (wALS) was expressed in the bacteria as a protein fused with glutathione S-transferase (GST). The fusion product GST-wALS was purified in a single step on a glutathione-Sepharose column. The purified GST-wALS was sensitive to a sulfonylurea herbicide, and was lost its sensitivity to end products, L-valine, L-leucine and L-isoleucine. These results suggest that the purified recombinant tobacco ALS was functionally active, and that the sulfonylureas may not bind to the feedback regulatory site on the plant ALS.