Helical sidedness and the distribution of polar residues in trans-membrane helices.
Trans-membrane helices have often been claimed to show "sidedness" in the distribution of polar and hydrophobic residues. However, an analysis of the statistical distribution of polar residues in randomly generated helices shows that the degree of bias commonly observed in real helices is far from statistically significant. It is concluded that "patchy" distributions of residues in such helices should be interpreted with great care.


Validity of the mandibular canine index (MCI) in sex prediction: Reassessment in an Indian sample.
The dentition's use in sex assessment has been explored and advocated owing to its strength and resistance to peri- and postmortem insults. Among the teeth, canines have shown the greatest sexual dimorphism; they are also known for greater resistance to pathological alterations and trauma. These factors have prompted some researchers to explore its use alone in sex prediction. Specifically the mandibular canine index (MCI), which is the ratio of the mesiodistal (MD) dimension of the lower right canine and the inter-canine arch width, was developed by Rao et al. [Forensic Sci. Int. 42 (1989) 249-254] who predicted sex correctly in ∼86% of cases in a South Indian sample. These results were verified subsequently by other Indian researchers, both on southern and on northern Indians, although such success rates were not apparent in another South-Asian population as well as a European group. The present study re-examined the method in 203 Indians (103 males, 100 females) of age 19-32 years who originated from diverse states of the country, constituting all major religious and caste affiliations, and found relatively low sex estimation accuracy using the MCI (∼51%). This is in spite of the MD dimension of canines as well as inter-canine arch width - the absolute measurements used for deriving the MCI - showing statistically significant sex differences (p<0.05); also, the application of multivariate statistics, viz. logistic regression analysis, revealed higher accuracy levels of sex estimation for the absolute measurements (∼62-66%) vis-à-vis the MCI (∼50%). The results herein reinforce recent observations that ratios obtained from teeth, such as the MCI, do not reflect sexual dimorphism that may be present in absolute measurements. Furthermore, we believe that the basis for using MCI as a sex assessment tool is questionable, since it depends on the levels of sexual dimorphism between the absolute dimensions-low sexual dimorphism in one (e.g. MD measurement of canines) and proportionately high sexual differences in the other (e.g. inter-canine arch width), or vice versa, would lead to higher sex estimation accuracy of the MCI; on the other hand, relatively equal levels of sexual dimorphism in both would negate the ability of the MCI to accurately estimate sex. These suggest that the MCI has little utility in sex estimation and that its application should be restricted, if not discontinued altogether, in forensic and anthropological sex prediction.


Computational proteomics analysis of binding mechanisms and molecular signatures of the HIV-1 protease drugs.
Computational proteomics analysis of biomolecular interactions is proposed to determine molecular signatures of the HIV-1 protease inhibitors. A comparative microscopic analysis is conducted for a panel of inhibitors which exemplify a diversity of the HIV-1 PR binding mechanisms, from the active site inhibition to intervening with the protease folding and dimerization. Replica-exchange Monte Carlo simulations with the conformational ensembles of the HIV-1 PR dimer and monomer structures enable a molecular analysis underlying diversity of the HIV-1 PR binding mechanisms. We have investigated the molecular basis underlying diversity of the HIV-1 PR binding mechanisms. The molecular basis of the HIV-1 PR active site and dimerization inhibition mechanisms has been analyzed for an active site tripeptide inhibitor and a tetrapeptide inhibitor, which can act as both a dimerization inhibitor and a competitive active site inhibitor. We have also simulated a structural mimicry mechanism of the HIV-1 PR folding inhibition and dimerization, according to which the folding inhibitor targets the conserved HIV-1 PR regions by mimicking the interaction network of the active dimer. We have shown that binding interfaces of the studied dimerization and folding HIV-1 PR inhibitors may enable structural mimicry with the hot spot residues of the HIV-1 PR dimer. The proposed structural models of intervening with the HIV-1 PR dimerization and folding support the mechanism of structural mimicry, which may alleviate drug resistance effects.


In vivo expansion of CD4+Foxp3+ regulatory T cells mediated by GITR molecules.
CD4(+)Foxp3(+) regulatory T cells (Treg cells) play an important role in maintaining self-tolerance as suppressive/regulatory CD4 T cells. In vitro analyses have revealed the characteristics of Treg cells, that is, hyporesponsiveness when stimulated via TCR in the presence of splenic APC. In this study, we report a new mAb, G3c, which can induce the expansion of Treg cells stimulated with anti-CD3 Ab along with splenic APC, the culture conditions in which Treg cells exhibit hyporesponsiveness. Surprisingly, G3c mAb recognized glucocorticoid-induced TNFR family-related proteins (GITR). G3c mAb had stronger co-stimulatory activity for both Treg cells and responder T cells than another anti-GITR Ab (DTA1) in vitro. The in vivo administration of G3c increased the number of Treg cells and had less effect in inducing anti-tumor immunity in normal mice, although G3c had some anti-tumor effect on non-Treg cells in the absence of Treg cells in vivo, in contrast to the anti-tumor therapeutic effect of DTA1 in normal mice. Therefore, we need to know that the manipulation of immune responses with the use of anti-GITR Abs results from a balance between co-stimulatory effects on Treg cells and on responder cells, and we must aim at a narrow window leading to the therapeutic effects.


The Role of ARF6 in Biliary Atresia.
Altered extrahepatic bile ducts, gut, and cardiovascular anomalies constitute the variable phenotype of biliary atresia (BA). To identify potential susceptibility loci, Caucasian children, normal (controls) and with BA (cases) at two US centers were compared at >550000 SNP loci. Systems biology analysis was carried out on the data. In order to validate a key gene identified in the analysis, biliary morphogenesis was evaluated in 2-5-day post-fertilization zebrafish embryos after morpholino-antisense oligonucleotide knockdown of the candidate gene ADP ribosylation factor-6 (ARF6, Mo-arf6). Among 39 and 24 cases at centers 1 and 2, respectively, and 1907 controls, which clustered together on principal component analysis, the SNPs rs3126184 and rs10140366 in a 3' flanking enhancer region for ARF6 demonstrated higher minor allele frequencies (MAF) in each cohort, and 63 combined cases, compared with controls (0.286 vs. 0.131, P = 5.94x10-7, OR 2.66; 0.286 vs. 0.13, P = 5.57x10-7, OR 2.66). Significance was enhanced in 77 total cases, which included 14 additional BA genotyped at rs3126184 only (p = 1.58x10-2, OR = 2.66). Pathway analysis of the 1000 top-ranked SNPs in CHP cases revealed enrichment of genes for EGF regulators (p<1 x10-7), ERK/MAPK and CREB canonical pathways (p<1 x10-34), and functional networks for cellular development and proliferation (p<1 x10-45), further supporting the role of EGFR-ARF6 signaling in BA. In zebrafish embryos, Mo-arf6 injection resulted in a sparse intrahepatic biliary network, several biliary epithelial cell defects, and poor bile excretion to the gall bladder compared with uninjected embryos. Biliary defects were reproduced with the EGFR-blocker AG1478 alone or with Mo-arf6 at lower doses of each agent and rescued with arf6 mRNA. The BA-associated SNPs identify a chromosome 14q21.3 susceptibility locus encompassing the ARF6 gene. arf6 knockdown in zebrafish implicates early biliary dysgenesis as a basis for BA, and also suggests a role for EGFR signaling in BA pathogenesis.


Lymphomagenesis-related gene expression in B cells from sustained virological responders with occult hepatitis C virus infection.
The expression of activation-induced cytidine deaminase, B-aggressive lymphoma, cyclin D1 and serine/threonine kinase 15 genes, among others, is increased in B cells from patients with chronic hepatitis C virus (HCV) infection. It is unknown whether the level of expression of these genes in B cells is increased in patients with hepatitis C who have achieved a sustained virological response (SVR) but who have persistent, detectable HCV RNA, so-called occult infection. Eighty-three patients who achieved and SVR, 27 with detectable HCV and 56 without detectable HCV RNA, 28 chronic hepatitis C patients and 32 healthy controls were studied. RNA was extracted from B cells, and gene expression levels were measured by RT-PCR. Patients with chronic HCV and those who achieved an SVR (with and without persistent low-level HCV RNA) showed a statistically significant higher expression compared to healthy controls, of activation-induced cytidine deaminase (P = 0.004, P < 0.001 and P = 0.002, respectively), B-aggressive lymphoma (P < 0.001, P = 0.001 and P = 0.006) and cyclin D1 (P = 0.026, P = 0.001; P = 0.038). For activation-induced cytidine deaminase patients with an SVR and 'occult infection' had a statistically significantly higher expression level than patients with and SVR without 'occult infection' (P = 0.014). The higher expression levels found for activation-induced cytidine deaminase, together with other genes indicates that these B lymphomagenesis-related genes are upregulated following HCV therapy and this is more marked when HCV can be detected in PBMCs.


Further characterization of preproenkephalin mRNA-containing cells in the rodent globus pallidus.
The globus pallidus (external pallidum of primates) is an essential nucleus within basal ganglia circuitry, in part because it receives at least one-half of striatal efferent projections. Neurons of the globus pallidus can be divided into subpopulations based on anatomical, physiological, and chemical features. Globus pallidus neurons project to several structures (the striatum, subthalamic nucleus, entopeduncular nucleus, and substantia nigra pars reticulata), have one of two alternative waveforms (positive/negative versus negative/positive), contain either the calcium binding protein parvalbumin or the neuropeptide precursor preproenkephalin mRNA and show differential immediate early gene responses to dopamine receptor agonists and antagonists. The objective of the present study was to characterize in greater detail the preproenkephalin mRNA-containing pallidal neurons using Sprague-Dawley rats. In situ hybridization for preproenkephalin mRNA was combined with immunocytochemical detection of: (i) the neuron-specific nuclear protein, NeuN, (ii) FluoroGold-labeled pallidostriatal and pallidosubthalamic cells, or (iii) Fos induced by either systemic combined D1-class/D2-class dopamine receptor agonists or a D2-class receptor antagonist. These experiments demonstrated that a substantial population (42%) of globus pallidus neurons contains preproenkephalin mRNA, and that globus pallidus neurons retrogradely labeled after FluoroGold injections into the striatum are more frequently preproenkephalinergic, compared to the population of pallidosubthalamic neurons. Furthermore, systemic administration of a D2 receptor antagonist, eticlopride, induced Fos immunoreactivity predominantly in globus pallidus neurons expressing preproenkephalin mRNA, while combined administration of D1 and D2 receptor agonists induced Fos predominantly in pallidal neurons lacking preproenkephalin mRNA.These results support the conclusion that preproenkephalin mRNA identifies one of the two major subpopulations of pallidal neurons. This preproenkephalin mRNA-expressing pallidal subpopulation preferentially targets the striatum and is more readily activated in its immediate early gene expression by D2 receptor antagonists than by dopamine receptor agonists. This projection provides a pallidal substrate for the dopaminergic regulation of striatal information processing.


Neonatal herpes simplex infection possibly acquired via maternal breast milk.
A newborn infant with disseminated herpes simplex virus type 1 (HSV-1) infection was determined serologically to have acquired the infection postnatally; his mother was found to have HSV-1 in her breast milk but had no history of genital lesions and negative viral cultures of cervix, vagina, and throat. We suggest that HSV-infected maternal breast milk may be a source of this infection for susceptible infants.


Estimating the Effect and Economic Impact of Absenteeism, Presenteeism, and Work Environment-Related Problems on Reductions in Productivity from a Managerial Perspective.
The aim of this study was to propose wage multipliers that can be used to estimate the costs of productivity loss for employers in economic evaluations, using detailed information from managers. Data were collected in a survey panel of 758 managers from different sectors of the labor market. Based on assumed scenarios of a period of absenteeism due to sickness, presenteeism and work environment-related problem episodes, and specified job characteristics (i.e., explanatory variables), managers assessed their impact on group productivity and cost (i.e., the dependent variable). In an ordered probit model, the extent of productivity loss resulting from job characteristics is predicted. The predicted values are used to derive wage multipliers based on the cost of productivity estimates provided by the managers. The results indicate that job characteristics (i.e., degree of time sensitivity of output, teamwork, or difficulty in replacing a worker) are linked to productivity loss as a result of health-related and work environment-related problems. The impact of impaired performance on productivity differs among various occupations. The mean wage multiplier is 1.97 for absenteeism, 1.70 for acute presenteeism, 1.54 for chronic presenteeism, and 1.72 for problems related to the work environment. This implies that the costs of health-related and work environment-related problems to organizations can exceed the worker's wage. The use of wage multipliers is recommended for calculating the cost of health-related and work environment-related productivity loss to properly account for actual costs.


Viral and non-viral vectors for cancer gene therapy.
Our research has focused on developing improved delivery vectors for treating cancer by gene therapy using the tumor suppressor p53 gene. Recombinant viral and non-viral vectors were used to deliver the p53 gene into non-small cell lung cancer (NSCLC) cells either in culture or as a subcutaneous tumor. Transduction of tumor cells was measured by beta-gal expression while tumor cell proliferation was used to measure the effect of p53. High level transduction was obtained in vitro and in vivo with a recombinant adenoviral vector, resulting in tumor cell growth inhibition in both models. A targeted, non-viral gene delivery vector based on the use of an EGF/DNA polyplex also resulted in efficient (as high as 66% transduction) and specific gene delivery in vitro when replication defective adenovirus was used as an endosome release agent. These vectors now provide improved methods to deliver therapeutic genes for cancer treatment by gene therapy.


Infective endocarditis presenting as polyarthritis.
We report the case of a patient who complained of arthralgias and arthritis 1 month before the onset of fever or other signs of infective endocarditis. In 2 months she developed an additive, asymmetrical polyarthritis with fever (febrile polyarthritis). Splenomegaly was present. Two-dimensional echocardiography showed no vegetations or other findings suggesting endocardial involvement. Initially, four blood cultures showed no microorganisms, then six of nine subsequent blood cultures grew highly gentamicin-resistant Enterococcus faecalis.


Synergic vascular photodynamic activity by methylene blue-curcumin supramolecular assembly.
A supramolecular assembly was obtained by combining methylene blue (MB) with a natural plant extract, curcumin (Curc), in a stoichiometric ratio of 1:4 in aqueous solution (90% PBS + 10% ethanol) at room temperature. The MB-Curc supramolecular assembly was evidenced by absorption and fluorescence spectroscopies, and the stoichiometry and bonding constant were obtained using Cieleńs model. Its stability and photostability were evaluated by chromatographic analysis and UV-Vis absorption. The MB-Curc avoids the aggregation of both isolated compounds and efficiently produces singlet oxygen (ΦΔ= 0.52 ± 0.03). Its potential for photodynamic antiangiogenic treatments was evaluated through the vascular effect observed in chicken chorioallantoic membrane (CAM) assay. The results showed intense damage in CAM vascular network by MB-Curc after irradiation, which is higher than the effect of isolated compounds, indicating a synergistic vascular effect. This combination can be essential to prevent cancer revascularization after photodynamic application and improve the efficacy of this approach. The characteristics exhibited by MB-Curc make it a potential candidate for use in cancer treatments through photodynamic antiangiogenic therapy.


Novel method utilizing bisulfite conversion with dual amplification-refractory mutation system polymerase chain reaction to detect circulating pancreatic β-cell cfDNA.
Several research groups have reported methods for quantifying pancreatic beta cell (β-cell) injury by measuring β-cell-specific CpG unmethylation of the insulin gene in circulation using digital droplet PCR or next-generation sequencing. However, these methods have certain disadvantages, such as the need to consider the background signal owing to the small number of target CpG sites and the need for unique equipment. We established a novel method for detecting four CpG unmethylations of the insulin gene using two-step amplification refractory mutation system PCR. We applied it to type 1 diabetes (T1D) patients with a wide range of disease durations and to healthy adults. The assay showed high linearity and could detect a single copy of unmethylated insulin DNA in experiments using methylated and unmethylated plasmid DNA. The unmethylated insulin DNA level in the type 1 diabetes group, whose β-cell mass was considerably reduced, was similar to that of healthy adults. An inverse correlation was observed between copy number and disease duration in patients with unmethylated insulin DNA-positive type 1 diabetes. We developed a novel method for detecting unmethylated insulin DNA in circulation that can be performed using a conventional real-time PCR system. This method would be useful for analyzing dynamic profiles of β-cells in human disease such as type 1 diabetes.


"The PLCP gene family of grapevine (Vitis vinifera L.): characterization and differential expression in response to Plasmopara Viticola".
Papain-like cysteine proteases (PLCPs), a large group of cysteine proteases, are structurally related to papain. The members belonging to PLCPs family contribute to plant immunity, senescence, and defense responses in plants. The PLCP gene family has been identified in Arabidopsis, rice, soybean, and cotton. However, no systematic analysis of PLCP genes has been undertaken in grapevine. Since Plasmopara viticola as a destructive pathogen could affect immunity of grapes in the field, we considered that the members belonged to PLCPs family could play a crucial role in defensive mechanisms or programmed cell death. We aimed to evaluate the role of PLCPs in 2 different varieties of grapevines and compared the changes of their expressions with the transcriptional data in response to P. viticola. In this study, 23 grapevine PLCP (VvPLCP) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Numerous cis-acting elements related to plant development, hormone, and stress responses were identified in the promoter of the VvPLCP genes. Phylogenetic analysis grouped the VvPLCP genes into nine subgroups. The transcription of VvPLCP in different inoculation time points and varieties indicated that VvPLCP may have vital functions in grapevine defense against Plasmopara viticola. According to transcriptome data and qPCR analysis, we observed the increasing expression levels of VvRD21-1 at 72 h after inoculation in resistant variety, inferring that it was related to grape downy mildew resistance. Meanwhile, 3 genes including VvXBCP1, VvSAG12-1, and VvALP1 showed higher expression at 24 h after pathogen inoculation in the susceptible variety and might be related to the downy mildew phenotype. We nominated these four genes to function during hypersensitive response (HR) process, inferring that these genes could be associated with downy mildew resistance in grapes. Our results provide the reference for functional studies of PLCP gene family, and highlight its functions in grapevine defense against P. viticola. The results help us to better understand the complexity of the PLCP gene family in plant immunity and provide valuable information for future functional characterization of specific genes in grapevine.


Angiopoietin-1 and -2 as markers for disease severity in hemolytic uremic syndrome induced by enterohemorrhagic Escherichia coli.
Angiopoietin (Ang)-1 and -2 play important roles in maintaining vascular homeostasis. This study aimed to assess the roles of angiopoietin (Ang)-1 and -2 and to investigate the clinical significance of their serum levels in patients with hemolytic uremic syndrome (HUS) induced by enterohemorrhagic Escherichia coli (EHEC). Twenty patients with HUS and 15 healthy controls were studied. Serum Ang-1 and Ang-2 levels were quantified using enzyme-linked immunosorbent assay. The results were compared with the clinical features of HUS. During the HUS phase, serum Ang-1 levels were significantly decreased, whereas serum Ang-2 levels and the Ang-2/Ang-1 ratio were significantly elevated. Compared with patients without encephalopathy, serum Ang-2 levels and Ang-2/Ang-1 ratio were significantly elevated in patients with encephalopathy. Patients with HUS and serum Ang-2 levels of >7061 pg/mL or Ang2/Ang1 ratios of >2.29 were at high risk of encephalopathy. Serum Ang-1 levels were significantly decreased in patients in the pre-HUS phase compared with those in healthy controls. Disruption of homeostasis of vascular endothelial function by Ang-1 and -2 may be closely associated with the development of HUS. Serum Ang-1 and -2 levels and the Ang-2/Ang-1 ratio may be promising indicators of disease activity in HUS and the development of encephalopathy.


Major impact of vitamin D3 deficiency and supplementation on left ventricular torsional mechanics during dobutamine stress in uncomplicated type 2 diabetes.
Hypovitaminosis D is associated with the risk of diabetic complications. Its role in diabetic-related cardiac abnormalities remain poorly understood. We aimed therefore to evaluate the effect of vitamin D deficiency and supplementation on early left ventricular (LV) dysfunction in vitamin D deficient patients with uncomplicated T2D. Sixty-three consecutive T2D patients who had a diagnosis of vitamin D3 were prospectively recruited and allocated into 2 groups (25(OH)D < 20 ng/mL: VDD, >20 ng/mL VDND). Twenty-eight of them with 25(OH)D < 20 ng/mL benefited from a 3-month supplementation. At baseline and follow-up, after conventional echocardiography including evaluation of epicardial adipose tissue (EAT), both LV longitudinal (LS) and circumferential (CS) strains and rotation/twist mechanics were evaluated at rest and during dobutamine (DOB) stress. After treatment, T2D patients successfully normalized their 25(OH)D levels. The strongest associations between vitamin D deficiency and supplementation with LV myocardial function were noticed for torsional mechanics indexes under DOB. EAT correlated significantly (p < 0.01) with baseline 25(OH)D and was reduced after supplementation. Significant correlations were obtained between these 2 parameters with twist or apical rotation at baseline (p < 0.01) and between their delta changes at follow-up (p < 0.01) under DOB. Significant improvements in LS and CS (p < 0.05) under DOB were also underlined at follow-up, with major enhancements noticed in the apical region (p < 0.01) of the LV. This study provides the first evidences of the potential of vitamin D supplementation as an efficient prophylactic strategy to alleviate the progression of myocardial dysfunction in asymptomatic patients with uncomplicated T2D. NCT03437421.


Multifunctional hybrid hydrogel with transparency, conductivity, and self-adhesion for soft sensors using hemicellulose-decorated polypyrrole as a conductive matrix.
Polymers with high conductivity and cross-linking ability are ideal materials for the preparation of conductive hydrogels for application in wearable electronic devices. However, the fabrication of conductive polymer-incorporated hydrogels with good synergistic properties remains a great challenge due to the hydrophobicity and opacity of conjugated π conductive polymers. In this study, a multifunctional hybrid hydrogel was prepared by incorporating hemicellulose-decorated polypyrrole (H/PPY), polyvinyl alcohol (PVA), and tannic acid (TA) into a polyacrylamide (PAM) network. The addition of excess ammonium persulfate (APS) in the process of gelation not only initiated the polymerization of PAM but also resulted in the change of the hydrogel from opaque to transparent by continuously breaking and reducing the size of the PPY particles. The hybrid hydrogel exhibited high transparency and conductivity, good adhesion ability and mechanical performance, and high resistance strain sensitivity and could accurately monitor the strain signals of the index finger and elbow flexion and pulse beat during rest and exercise, which has promising potential for use in wearable or implantable smart sensor devices, electronic skins, and artificial intelligence applications.


Estradiol assay by microtitre plate enzyme immunoassay.
Development of a simple enzyme linked immunosorbent assay (ELISA) for estradiol in serum extracts is described. The assay involves use of a 96-well microtitre plate, designed for immunoassay, as the support for a purified, high-titre antiserum, raised against estradiol-6(O)-carboxymethyloxime linked to bovine serum albumin, and using horseradish peroxidase-labelled estradiol-6-(O)-carboxymethyloxime as the labelled species, with 2,2'-azino-bis-(3-ethylbenzthiazoline sulfonic acid) diammonium salt (ABTS) as the chromogenic substrate. The assay characteristics rival those of radio- or chemiluminescence immunoassays for estradiol.


Battlefield acupuncture instead of opioids for abscess drainage in the pediatric emergency department.
Procedural analgesia with opioids is often needed during incision and drainage (I/D) of pilonidal or large abscesses in the pediatric emergency department (PED). In response to the ongoing opioid misuse pandemic, the Joint Commission has mandated that nonpharmacologic therapies be offered to patients for pain first. Battlefield Acupuncture (BFA) is an auricular acupuncture protocol that is drug free, can achieve rapid pain relief, and is used in the United States military. BFA for painful procedures in the PED has not been reported. We describe 4 cases where BFA was utilized in a PED for pain during I/D of abscesses.


Sensitivity to orthographic vs. phonological constraints on word recognition: An ERP study with deaf and hearing readers.
The role of phonology in word recognition has previously been investigated using a masked lexical decision task and transposed letter (TL) nonwords that were either pronounceable (barve) or unpronounceable (brvae). We used event-related potentials (ERPs) to investigate these effects in skilled deaf readers, who may be more sensitive to orthotactic than phonotactic constraints, which are conflated in English. Twenty deaf and twenty hearing adults completed a masked lexical decision task while ERPs were recorded. The groups were matched in reading skill and IQ, but deaf readers had poorer phonological ability. Deaf readers were faster and more accurate at rejecting TL nonwords than hearing readers. Neither group exhibited an effect of nonword pronounceability in RTs or accuracy. For both groups, the N250 and N400 components were modulated by lexicality (more negative for nonwords). The N250 was not modulated by nonword pronounceability, but pronounceable nonwords elicited a larger amplitude N400 than unpronounceable nonwords. Because pronounceable nonwords are more word-like, they may incite activation that is unresolved when no lexical entry is found, leading to a larger N400 amplitude. Similar N400 pronounceability effects for deaf and hearing readers, despite differences in phonological sensitivity, suggest these TL effects arise from sensitivity to lexical-level orthotactic constraints. Deaf readers may have an advantage in processing TL nonwords because of enhanced early visual attention and/or tight orthographic-to-semantic connections, bypassing the phonologically mediated route to word recognition.


Canine susceptibility to visceral leishmaniasis: A systematic review upon genetic aspects, considering breed factors and immunological concepts.
Dogs have different susceptibility degrees to leishmaniasis; however, genetic research on this theme is scarce, manly on visceral form. The aims of this systematic review were to describe and discuss the existing scientific findings on genetic susceptibility to canine leishmaniasis, as well as to show the gaps of the existing knowledge. Twelve articles were selected, including breed immunological studies, genome wide associations or other gene polymorphism or gene sequencing studies, and transcription approaches. As main results of literature, there was a suggestion of genetic clinical resistance background for Ibizan Hound dogs, and alleles associated with protection or susceptibility to visceral leishmaniasis in Boxer dogs. Genetic markers can explain phenotypic variance in both pro- and anti-inflammatory cytokines and in cellular immune responses, including antigen presentation. Many gene segments are involved in canine visceral leishmaniasis phenotype, with Natural Resistance Associated Macrophage Protein 1 (NRAMP1) as the most studied. This was related to both protection and susceptibility. In comparison with murine and human genetic approaches, lack of knowledge in dogs is notorious, with many possibilities for new studies, revealing a wide field to be assessed on canine leishmaniasis susceptibility research.


Risk factors of short-term mortality after acute nonvariceal upper gastrointestinal bleeding in patients on dialysis: a population-based study.
Impaired kidney function is an established predictor of mortality after acute nonvariceal upper gastrointestinal bleeding (ANVUGIB); however, which factors are associated with mortality after ANVUGIB among patients undergoing dialysis is unknown. We examined the associations among demographic characteristics, dialysis-specific features, and comorbid conditions with short-term mortality after ANVUGIB among patients on dialysis. Retrospective cohort study. United States Renal Data System (USRDS), a nation-wide registry of patients with end-stage renal disease. All ANVUGIB episodes identified by validated algorithms in Medicare-covered patients between 2003 and 2007. Demographic characteristics and comorbid conditions from 1 year of billing claims prior to each bleeding event. We used logistic regression extended with generalized estimating equations methods to model the associations among risk factors and 30-day mortality following ANVUGIB events. From 2003 to 2007, we identified 40,016 eligible patients with 50,497 episodes of ANVUGIB. Overall 30-day mortality was 10.7% (95% CI: 10.4-11.0). Older age, white race, longer dialysis vintage, peritoneal dialysis (vs. hemodialysis), and hospitalized (vs. outpatient) episodes were independently associated with a higher risk of 30-day mortality. Most but not all comorbid conditions were associated with death after ANVUGIB. The joint ability of all factors captured to discriminate mortality was modest (c=0.68). We identified a profile of risk factors for 30-day mortality after ANVUGIB among patients on dialysis that was distinct from what had been reported in non-dialysis populations. Specifically, peritoneal dialysis and more years since initiation of dialysis were independently associated with short-term death after ANVUGIB.


Perceived proficiency in endoscopic techniques among senior obstetrics and gynecology residents.
To assess current training methods in laparoscopic surgery employed in United States obstetrics and gynecology residency programs, level of proficiency in various minimally invasive surgery procedures amongst senior obstetrics and gynecology residents, and ways in which training in minimally invasive surgery can be improved. Survey (Canadian Task Force classification III). Accredited obstetrics and gynecology programs in the United States. All fourth-year residents in accredited obstetrics and gynecology programs in the United States. Residents received a survey regarding their perceived proficiency performing various laparoscopic procedures and the type of training they received in these techniques. Responses were received from 133 programs (52.4%) and 295 residents (26.8%). Of these, 67% of residents thought emphasis on laparoscopic surgery training should be increased or greatly increased; 87% thought laparoscopic skills were important for building a successful practice. Formal teaching methods were clearly associated with improved perception of proficiency, and those with higher perception of proficiency expected to perform more laparoscopic procedures after graduation. Residents lacked perceived competency in most advanced laparoscopic procedures. Residents seem to benefit significantly from a formal curriculum in minimally invasive surgery, but they do not feel competent performing some advanced procedures on graduation. In our opinion, more emphasis should be placed on training in laparoscopic surgery in United States obstetrics and gynecology programs.


Chronic hematic cyst of the orbit.
A chronic hematic cyst of the orbit caused unilateral globe displacement in four patients, two of whom had a past history of orbital trauma. The onset of a painless vertical displacement of the globe ranged from 3 weeks to 4 years. Computed tomographic (CT) findings demonstrated a homogeneous, nonenhancing, subperiosteal mass in the superior orbit in three patients and the inferior orbit in one. At surgery, all patients had masses surrounded by a pseudocapsule without epithelial or endothelial lining. The history, age distribution, and CT findings help to differentiate this entity from other causes of unilateral exophthalmos.


Implantable gastric stimulation (IGS) as therapy for human morbid obesity: report from the 2001 IFSO symposium in Crete.
This supplement contains reports from a symposium on a novel approach to treat obesity, gastric myo-electrical stimulation, that was held at the IFSO in Greece in 2001. There were four presentations. Xavier Pi-Sunyer from Columbia University in New York discussed medical risks of obesity. Karl Miller from Austria presented technical aspects of the surgery. Valerio Cigaina from Italy, the originator of the concept, reviewed his 7-year results with this therapy. Finally, Jerome D'Argent from France gave his preliminary results employing higher energy electrical stimulation parameters. Customarily weight loss data are presented as percent excess weight lost (%EWL), an antiquated measurement, e.g. all subjects were purchasing life insurance, the poor and minorities were under-represented and those with heart disease, malignancies or diabetes were excluded. In this supplement weight loss data are presented in a novel manner: percent excess body mass index (BMI) lost %EBL. This innovation merits an explanation and justification. The NIH/NIDDK convened a panel, chaired by Professor Pi-Sunyer, that concluded that for adults a BMI of 25 should be considered the upper limit of normal. Accordingly, we have proposed that BMI units in excess of 25 be considered to represent 100% of the excess weight of an individual. The USA FDA has agreed to accept weight loss data presented as %EBL. The intriguing data presented justify further evaluation of this novel, potentially useful and relatively benign treatment of obesity.


Alteration of erythrocytes by reduced species of oxygen generated in hypoxic rat liver during perfusion.
Activated species of oxygen produced in the hypoxic rat liver leave the cells and attack the membrane of circulating erythrocytes. Therefore the metabolic and structural changes of red blood cells may be appropriate indicators of an oxidative loading in tissues. The circulating erythrocytes exert a profound protective, "radical scavenger effect" on the liver tissue.


Time-dependent effects of actinomycin S3 on sleep and brain concentrations of 5-hydroxytryptamine in the rat.
Changes in the sleep pattern were investigated in rats given actinomycin S3 (AcS3, 0.25 mg/kg, intraventricularly), an inhibitor of RNA synthesis, at 0000, 0600, 1200, and 1800. The amounts of slow-wave (SS) and paradoxical sleep (PS) were increased during the dark period (1800 to 0600) in the groups treated at 0000, 0600, and 1200. In the 1800 injected group, PS was decreased. 5-Hydroxytryptamine (5-HT) was determined in the 0600 injected group. The 5-HT concentration in the AcS3-treated group was twofold higher than in the control group at 1800 (12 h after administration) in the cerebral cortex and lower brain stem but returned to control values at 0000 (18 h after) while sleep time continued to increase. Thus, AcS3 affected the circadian rhythm of the sleep-wake cycle. The increase of sleep time in the dark period was independent of changes in brain 5-HT content.


Effects of dry needling on symptomatic hallux valgus: A randomized single blind clinical trial.
The purpose of the present study was to determine the short and medium effects of dry needling (DN) on myofascial trigger points (MTrPs) in individuals with symptomatic hallux valgus (SHV). A total of 30 female volunteers, aged 25-60, with hallux valgus (HV) were randomly divided into two groups: DN group (n = 15) and control group (n = 15) who received sham dry needling. The outcome measures were pain intensity (Visual Analogue Scale, VAS), Foot Function Index (FFI), hallux valgus angle (HVA), and first metatarsophalangeal joint radiography. At the end of the intervention, the HVA showed a significant decrease in the case group (P < 0.001); however, statistically no significant difference was found in the pain intensity and foot function between the two groups (P > 0.05). These findings were maintained for a week and a month during follow-ups. According to the findings, dry needling can be recommended for improving first metatarsophalangeal joint alignment in the mild to moderate SHV individuals.


Angiographic evaluation of the coronary vasodilating effect of trapidil in anesthetized closed-chest dogs.
We studied the effect of trapidil on coronary diameters of anesthetized closed-chest dogs using coronary angiography. Trapidil 2 mg/kg i.v. significantly (p less than 0.01) increased the diameter of the proximal (1.96 +/- 0.08 mm), middle (1.29 +/- 0.05 mm), and distal (0.86 +/- 0.03 mm) coronary arteries by 4.2 +/- 1.0, 7.6 +/- 1.2, and 10.8 +/- 2.2%, respectively. Nitroglycerin 20 micrograms/kg i.v. significantly (p less than 0.01) increased the diameter of classes I, II, and III arteries by 4.0 +/- 0.7, 7.7 +/- 1.3, and 7.5 +/- 1.8%, respectively. Ergonovine (0.4 mg i.v.; n = 13) reduced (p less than 0.01) the diameter of all three classes of coronary arteries by 8.2 +/- 1.2, 9.0 +/- 1.0, and 13.3 +/- 1.9%. After pretreatment with ergonovine, trapidil (n = 8) relaxed the coronary arteries from -10.4 +/- 1.7 to -2.4 +/- 1.7% constriction (p less than 0.05). These results indicate that in dogs trapidil dilates coronary arteries in a fashion similar to nitroglycerin and reverses ergonovine-induced coronary vasoconstriction.


Alternative drugs against multiresistant Gram-negative bacteria.
Enterobacterales and other non-fermenting Gram-negative bacteria have become a threat worldwide owing to the frequency of multidrug resistance in these pathogens. On the other hand, efficacious therapeutic options are quickly diminishing. The aims of this study were to describe the susceptibility of 50 multiresistant Gram-negative bacteria, mostly pan-resistant, against old and less-used antimicrobial drugs and to investigate the presence of antimicrobial resistance genes. A total of 50 genetically distinct isolates were included in this study, including 14 Acinetobacter baumannii (belonging to ST79, ST317, ST835 and ST836), 1 Pseudomonas aeruginosa (ST245), 8 Serratia marcescens and 27 Klebsiella pneumoniae (belonging to ST11, ST340, ST258, ST16, ST23, ST25, ST101, ST234, ST437 and ST442). The isolates were submitted to antimicrobial susceptibility testing and whole-genome sequencing to evaluate lineages and resistance genes. Our results showed that some strains harboured carbapenemase genes, e.g. blaKPC-2 (28/50; 56%) and blaOXA-23 (11/50; 22%), and other resistance genes encoding aminoglycoside-modifying enzymes (49/50; 98%). Susceptibility rates to tigecycline (96%) in all species (except P. aeruginosa), to minocycline (100%) and doxycycline (93%) in A. baumannii, to ceftazidime/avibactam in S. marcescens (100%) and K. pneumoniae (96%), and to fosfomycin in S. marcescens (88%) were high. Chloramphenicol and quinolones (6% susceptibility each) did not perform well, making their use in an empirical scenario unlikely. This study involving genetically distinct bacteria showed promising results for tigecycline for all Gram-negative bacteria (except P. aeruginosa), and there was good activity of minocycline against A. baumannii, ceftazidime/avibactam against Enterobacterales, and fosfomycin against S. marcescens.


Identification of microRNA precursor with the degenerate K-tuple or Kmer strategy.
The microRNA (miRNA), a small non-coding RNA molecule, plays an important role in transcriptional and post-transcriptional regulation of gene expression. Its abnormal expression, however, has been observed in many cancers and other disease states, implying that the miRNA molecules are also deeply involved in these diseases, particularly in carcinogenesis. Therefore, it is important for both basic research and miRNA-based therapy to discriminate the real pre-miRNAs from the false ones (such as hairpin sequences with similar stem-loops). Most existing methods in this regard were based on the strategy in which RNA samples were formulated by a vector formed by their Kmer components. But the length of Kmers must be very short; otherwise, the vector's dimension would be extremely large, leading to the "high-dimension disaster" or overfitting problem. Inspired by the concept of "degenerate energy levels" in quantum mechanics, we introduced the "degenerate Kmer" (deKmer) to represent RNA samples. By doing so, not only we can accommodate long-range coupling effects but also we can avoid the high-dimension problem. Rigorous jackknife tests and cross-species experiments indicated that our approach is very promising. It has not escaped our notice that the deKmer approach can also be applied to many other areas of computational biology. A user-friendly web-server for the new predictor has been established at http://bioinformatics.hitsz.edu.cn/miRNA-deKmer/, by which users can easily get their desired results.


On the optimal choice of the electrode number and locations in body surface mapping.
The problem of choosing the number and the thoracic locations of the leads needed for reconstructing body surface maps of heart potentials is considered. In a previous work (P. Barone, In "Computers in Cardiology" (K. L. Ripley, Ed.), pp. 491-494. IEEE Computer Soc. Press, Washington, DC, 1985) the problem was solved, provided that the user is able to give a threshold below which the information content of two leads is considered equal. In this paper a sensitivity analysis that can help the user in choosing the optimal threshold, or, equivalently, the optimal number of leads, is performed. The problem is reduced to the dynamic search of spanning forests of an undirected graph. An example using real data is also discussed.


Nonlinear optical effects related to saturable and reverse saturable absorption by subphthalocyanines at 532 nm.
It is found that both effects of saturable absorption and reverse saturable absorption are obtained with a solution of subphthalocyanine at 532 nm depending on the intensity of 9 ns laser pulses; saturable absorption occurs at lower intensity levels whereas the reverse effect prevails at higher levels; contrary to expectations, subphthalocyanines can behave as reverse saturable absorbers at 532 nm, despite the high linear absorption at this wavelength; data have been fitted with a five-level model which considers three consecutive electronic transitions with absorption cross-section values of 1.4 x 10(-16), 1.0 x 10(-16) and 40 x 10(-16) cm(2), respectively.


Survey of clinician opinions on the role of proton beam therapy in Australia and New Zealand.
We surveyed the Australian and New Zealand (ANZ) radiation oncology community to assess their perceptions, understanding and experience of the current role of proton beam therapy (PBT) and the existing referral process to access PBT overseas, ahead of the development of the first PBT centre in Australia. The survey was conducted between September and October 2019 using a 17-question instrument, which was distributed by email to all 632 radiation oncology fellows and trainees listed in the Royal Australian and New Zealand College of Radiologists database. One hundred and one respondents completed the survey, with an overall response rate of 16%. Most respondents were based in Australia (93%), with the majority working in public centres only (59%); 51% were > 10 years post fellowship and 17% were trainees. Most respondents (76%) reported moderate or high levels of confidence in the role of PBT. Only 28% had previously referred a patient for PBT overseas, with the most common referral indication being chordoma. Of those who had not previously referred a patient, 48% were not convinced about the rationale of PBT over current therapies available locally, 33% were not aware of the referral process, and 24% had concerns about the timeliness of a decision for government-funded PBT abroad. This survey has demonstrated that, although there is reasonable confidence in the role of PBT among ANZ radiation oncologists, there are a number of important aspects of PBT awareness, education and access that need to be developed prior to commencement of PBT in Australia.


Intrapericardial extralobar pulmonary sequestration presenting as a prenatal intrathoracic mass.
An intrathoracic mass, which persisted during the remaining pregnancy, was first seen during routine ultrasound examination performed at 20 weeks gestation. After birth, the child was asymptomatic. Echocardiography showed the mass to be located intrapericardially. The mass was electively resected via sternotomy 3 weeks after the birth. Microscopic examination showed normal lung tissue surrounded by pleura corresponding to the diagnosis of extralobar pulmonary sequestration. To the authors' knowledge, this is the earliest described detection of such a lesion. Furthermore, this article reports the unique finding of a feeding vessel from the right pulmonary artery.


Effect of manuka honey socket dressing on postoperative sequelae and complications following third molar extraction: A randomized controlled study.
To evaluate the effect of manuka honey on the healing of mandibular third molar extraction sockets. This was a prospective randomized study at a tertiary health institution in Lagos, Nigeria. All extractions were transalveolar, using the distobuccal bone guttering technique. Participants were randomized into two study groups. Group A underwent intrasocket application of manuka honey, after which sockets were completely closed using the mucosal flap, while participants in group B underwent mucosal flap closure of the socket without application of manuka honey. The primary outcome was healing of the extraction socket at 7 days postoperatively. The secondary outcomes measured were postoperative sequelae, namely pain, swelling, and trismus on postoperative days 1, 3, and 7, and socket healing complications - specifically inflamed socket, infected socket, and alveolar osteitis. In total, 112 participants completed the study, with 56 participants per group. There were no significant differences in demographic variables between both groups. On the 7th day postoperatively, 26.8% of participants in group B had an unhealed extraction site, compared with 10.3% of participants in group A (p = 0.029). A significant difference was observed between pre- and postoperative pain scores in both study groups (p = 0.001). A comparison of postoperative mean facial swelling between the two groups showed no statistically significant differences on all the review days (p = 0.66). The difference in postoperative socket healing complication rate between both groups was statistically significant (χ2 = 4.747, p = 0.029). Within the limitations of the study it seems that the application of manuka honey appears to aid earlier healing of the third molar extraction socket, with a significantly lower frequency of complications. Therefore, the application of manuka honey is recommended whenever appropriate.


Geometrid caterpillar in Eocene Baltic amber (Lepidoptera, Geometridae).
Lepidoptera have little fossilization potential due to the presence of delicate structures and hence are exceptional findings, even in ambers that allow their preservation in sufficient detail for interpretation. From Eocene Baltic amber, the volumetrically largest known deposit of amber, there has been no reliable report of any member of the Macrolepidoptera (informal group of higher moths and all butterflies). Any such lepidopteran fossil would provide insight into evolutionary processes during the Eocene, long after flowering plants had completed their initial radiation. Here, we report on a first geometrid caterpillar from Baltic amber which is described as the oldest evidence for the subfamily Ennominae (tribe Boarmiini) and as one of the oldest records of the currently mega-diverse family. The new finding provides an important calibration point for molecular clock analyses within the family Geometridae and predates the basal divergence of Boarmiini from 32-38 to 44 Mya. It also predates the occurrence of this highly specialized form of caterpillar locomotion that allows for rapid movement.


The natural history of renal function following orthotopic heart transplant.
The outcome of solid organ transplantation has dramatically improved after the introduction of the calcineurin inhibitor cyclosporine. With the increasing longevity of heart transplant recipients, the long-term effects of cyclosporine on renal function have become more evident. The natural history of kidney function following orthotopic heart transplant is not well defined and long-term follow up studies are scant. We conducted an observational study on patients who received a heart transplant at Saint Louis University Hospital between January 1, 1983 and December 31, 1988. Patients were followed up for 15 yr or until death whichever occurred first. In order to assess the effect of heart transplantation and cyclosporine exposure on long-term renal function we restricted the statistical analysis to patients who survived the first year post-transplantation. A total of 68 patients received orthotopic heart transplants at Saint Louis University Hospital between 1983 and 1988. Forty-eight (71%) patients survived for more than 1 yr. All patients were treated with cyclosporine based triple immunosuppressive regimen, with gradual cyclosporine dose reduction over time. The mean duration of follow-up was 8 yr. The estimated GFR at 5 and 10 yr post-transplant were significantly lower than estimated GFR at baseline and 1 yr post-transplant. There was no significant difference between estimated GFR at 15 yr and estimated GFR at baseline or 1 yr post-transplant. The cumulative incidence of chronic renal failure (GFR < or = 29 mL/min/1.73 m2) at 5, 10 and 15 yr was 4.2, 10.4 and 12.5%, respectively (p < 0.05). The cumulative incidence of severe chronic renal failure (GFR < or = 15 mL/min/1.73 m2) at 5, 10 and 15 yr was 2.1, 8.3 and 8.3%, respectively. The mortality rate was 8, 37, and 52% at 5, 10, and 15 yr, respectively. The 10 and 15 yr survivors had an estimated GFR at 1 yr post-transplant that was significantly higher than the non-survivors. Age, pre-transplantation estimated GFR, pre-transplantation diabetes and pre-transplantation hypertension are risk factors associated with > or = 10 mL/min/1.73 m2 decrement in estimated GFR. Heart transplant survivors beyond the first year post-transplant have a significant decrease in renal function and significant mortality observed over time. Age, pre-transplant GFR, pre-transplant diabetes and pre-transplant hypertension are important risk factors for decrement in renal function.


Restoration of micturition using microelectric current in experimentally induced spastic urinary bladder in rabbits.
The 29 rabbits used in this study were divided into three groups, A (A1 and A2), B, and C. In subgroup A1, 4 animals were used in order to verify whether the contact of an electrode to the sacral nerves results in some abnormality of voiding reflex. In subgroups A1 and A2 (4 animals each) we further studied the micturition function using three parameters: (i) urinary bladder fluoroscopy and radiography, (ii) cystomanometry, and (iii) electromyography of the pelvic floor muscles (external sphinter). In group B (9 rabbits) spastic paraplegia and micturition disturbances resulted from spinal cord compression that was induced by inserting a balloon catheter into the T11-T12 intervertebral foramen. In this group the parameters studied revealed a spastic urinary bladder in all animals. Finally, the 12 animals of group C were rendered paraplegic as described in group B, and microelectrodes were placed over the sacral nerves as in subgroup A1. By applying a specific sequence of sacral nerve stimulation we succeeded in satisfactory urinary bladder emptying as confirmed by the micturition parameters studied: The urinary bladder pressure decreased from 65 +/- 3 to 28 +/- 3 mm Hg. The pelvic floor muscle amplitude was lowered from 130 +/- 7 to 20 +/- 3 microV, and finally the radiological bladder size also decreased from 38 cm2 before voiding to 18 +/- 3 cm2 after voiding. These results indicate that microelectric current stimulation of the sacral nerves, when applied under a specific sequence, could rather satisfactorily restore micturition disturbances, at least in this experimental animal.


Reactivity patterns with HLA-A2 variants indicate lack of identity between determinants defined by monoclonal antibodies and cytotoxic-T-cell clones.
The anti-HLA-A2 monoclonal antibodies (MoAb) CR11-351 and 4B inhibit the binding of each other to HLA-A2 lymphoid cells and block the cytotoxicity of the anti-HLA-A2 cytotoxic-T-cell clone R32. The blocking does not reflect reactivity of the MoAb CR11-351 and 4B and of the cytotoxic-T-cell clone R32 with the same determinant, since they display differential reactivity with four HLA-A2 variants which carry amino acid substitutions at different positions. These results show for the first time in the human system that Class I HLA variants represent useful reagents to compare the fine specificities of monoclonal antibodies and T-cell clones. Furthermore our data suggest that T-cell recognition depends upon the tertiary structure of the antigen.


Implementing Integrated Community-Based Primary Healthcare: Applying the iCoach-Approach to Case Selection to Denmark.
The iCoach approach to case selection focuses on innovative models of community-based primary healthcare (CBPHC) and their contexts. The aim of this study was to assess the possibilities and limitations of the approach based on Denmark, which differs in significant ways from the jurisdictions initially included. Case study research suggests the approach is an interesting attempt to standardise case selection based on literal replication. The study reviewed the national grey literature and interviewed key informants at national and local levels. Applying the approach to Denmark required redefining selection criteria related to collaboration and context to capture its specific institutional and policy context. Selecting cases at the organisational level also required assessing how the system level contexts compared to those of the initial three jurisdictions included in iCoach. The iCoach approach allows collecting broadly comparable cases of innovative models of CBPHC across jurisdictions. However, the analysis of underlying conditions of implementing innovative models requires a more interactive approach to case selection. Researchers need to be clearer about the specific purpose of the case selection. This is also highly relevant for practitioners to ensure that insights are applicable in specific local and national contexts.


Multigenic control of thyroid hormone functions in the nervous system.
Thyroid hormone (TH) has a remarkable range of actions in the development and function of the nervous system. A multigenic picture is emerging of the mechanisms that specify these diverse functions in target tissues. Distinct responses are mediated by alpha and beta isoforms of TH receptor which act as ligand-regulated transcription factors. Receptor activity can be regulated at several levels including that of uptake of TH ligand and the activation or inactivation of ligand by deiodinase enzymes in target tissues. Processes under the control of TH range from learning and anxiety-like behaviour to sensory function. At the cellular level, TH controls events as diverse as axonal outgrowth, hippocampal synaptic activity and the patterning of opsin photopigments necessary for colour vision. Overall, TH coordinates this variety of events in both central and sensory systems to promote the function of the nervous system as a complete entity.


Blockade of chemo-resistance to 5-FU by a CK2-targeted combination via attenuating AhR-TLS-promoted genomic instability in human colon cancer cells.
As highly expressed in several human cancers, Casein Kinase 2 (CK2) is involved in chemotherapy-induced resistance. As a new potent CK2 inhibitor, DN701 is used to overcome chemoresistance through its synergistic antitumor effect with 5-fluorouracil (5-FU). Translesion DNA synthesis (TLS) has drawn our attention because it is associated with the development of chemo-resistance and tumor recurrence. The in vitro biological properties of 5-FU-resistant colon cancer cells revealed that DN701 combined with 5-FU could overcome chemo-resistance via blocking CK2-mediated aryl hydrocarbon receptor (AhR) and TLS-induced DNA damage repair (DDR). Moreover, pharmacologic and genetic inhibitions of AhR potently reduced TLS-promoted genomic instability. The mechanistic studies showed that combined DN701 with 5-FU was investigated to inhibit CK2 expression level and AhR-TLS-REV1 pathway. Meanwhile, DN701 combined with 5-FU could reduce CK2-AhR-TLS genomic instability, thus leading to superior in vivo antitumor effect. The insights provide a rationale for combining DN701 with 5-FU as a therapeutic strategy for patients with colon cancer.


Clinical trials update from the European Society of Cardiology Heart Failure meeting and the American College of Cardiology: darbepoetin alfa study, ECHOS, and ASCOT-BPLA.
This article provides information and a commentary on landmark trials presented at the European Society of Cardiology Heart Failure meeting held in June 2005, relevant to the pathophysiology, prevention and treatment of heart failure. All reports should be considered as preliminary data, as analyses may change in the final publication. The erythropoiesis stimulating protein, darbepoetin alfa, increased haemoglobin levels, improved quality of life and showed a trend for improved exercise duration in anaemic patients with symptomatic chronic heart failure. In the ECHOS study, the selective dopamine agonist nolomirole (CHF1035) showed no benefit in heart failure patients. Preliminary results of the ASCOT-BPLA study, which were reported at the American College of Cardiology meeting in March 2005, showed that in hypertensive patients, treatment with a calcium antagonist plus an ACE inhibitor was more effective at reducing cardiovascular outcomes than atenolol plus a diuretic.


Prestin as a biochemical marker for early detection of acquired sensorineural hearing loss.
Acquired sensorineural hearing loss and tinnitus can come about through various etiologies such as exposure to excessively loud noise or drugs with ototoxic properties. As such, acquired hearing loss is a common source of morbidity which deleteriously affects the ability to communicate. At present our ability to detect acquired hearing loss and tinnitus at its earliest stages is limited and there are no adjuncts to audiometric evaluation. The earliest cellular targets of noise and ototoxins in the cochlea are the outer hair cells (OHC). I hypothesize that serum assays of OHC specific protein, prestin, will allow detection and quantification of OHC damage before audiometric testing can identify presence of hearing loss. At present, there are no data available to evaluate this hypothesis, but initial evaluation can readily be carried out using existing experimental animal models of ototoxicity and noise-induced hearing loss. Early detection of OHC damage is critical to adoption of measures aimed at ameliorating hearing loss and tinnitus, thus reducing permanent deficits and disability.


A quantitative comparison analysis of diatoms in the lung tissues and the drowning medium as an indicator of drowning.
The presence of diatoms in the lung tissues, internal organs and bone marrow is considered as the supportive evidence in the diagnosis of death by drowning. Generally, the diatoms detected in the lung tissues are regarded as insignificant since these diatoms can be detected in the lung tissues of the postmortem immersion bodies. In this study, we analyzed the relationships between the numbers of the diatoms in the lung tissues and the drowning medium. We made a comparison analysis between the diatoms in the lung tissues and the drowning medium using the ratio of diatom numbers in both samples (L/D ratio), utilizing Microwave Digestion - Vacuum Filtration - Automated Scanning Electron Microscopy method. Our data indicate that the L/D ratios in victims of the drowning group were higher than the postmortem immersion group. A higher L/D ratio provides valuable information about the cause of death in drowning victims. Quantitative diatom analysis in the lung tissues, especially combined with the diatom analysis of the drowning medium, provides supportive evidence in determining if a body recovered in water was due to drowning or not.


Risk Factors for Female Breast Cancer: A Population Cohort Study.
The incidence of female BC among the Eastern and Southeastern Asian populations has gradually increased in recent years. However, epidemiological studies on the relationship between a sedentary lifestyle and female BC are insufficient. In order to determine the association between this lifestyle and the incidence of female BC, we conducted a population-based cohort study on women in Taiwan. We followed a prospective cohort of 5879 women aged 30 years and over enrolled in the 2001 National Health Interview Survey (NHIS), who developed female BC over a period of 72,453 person years, and we estimated the hazard ratios (HRs) and corresponding 95% confidence intervals (CIs) using the Cox proportional hazards model. RFs associated with female BC incidence included parity (adjusted HR = 0.63; 95% CI: 0.44-0.91), body mass index (adjusted HR = 1.34; 95% CI: 1.04-1.71), and ≥3 h/day spent sitting (adjusted HR = 1.89; 95% CI: 1.08-3.32). The incidence of female BC in participants who sat for ≥3 h/day and consumed sugary drinks was 2.5 times greater than that in those who sat for <3 h/day and did not consume sugary drinks (adjusted HR = 2.51; 95% CI: 1.01-6.23). The findings of this study indicate that sedentary behavior and sugary drink intake may increase the risk of developing female BC. These are modifiable RFs; therefore, a healthy lifestyle and diet can reduce the incidence of female BC.


Delayed diagnosis of a non-communicating right uterine horn: A case report and literature review.
Accurate diagnosis of a müllerian anomaly is essential for appropriate management and prevention of complications. However, diagnosis is often missed or delayed. This is a case of a nulliparous woman with a müllerian anomaly diagnosed at the age of 36 despite prior evaluation with ultrasound and laparoscopy. Magnetic resonance imaging (MRI) suggested a unicornuate uterus with a right non-communicating rudimentary horn. Hysteroscopy and chromopertubation confirmed the diagnosis. The rudimentary horn was resected laparoscopically using bipolar energy. An undiagnosed müllerian anomaly should be considered in the setting of persistent dysmenorrhea. Three-dimensional ultrasound or MRI should be used judiciously in patients with refractory dysmenorrhea or when ultrasound raises concern for a müllerian anomaly.


Electric field effects on the virus M13, detected by electro-optical measurements.
We have carried out transient electric birefringence experiments with M13 in aqueous solution. A normal effect with a negative sign is superimposed by anomalous positive effects. These prevail more and more upon raising the field strength. They apparently reflect electrically induced structural transitions. The threshold field strength decreases when buffer and/or electrolyte is removed.


The emerging causal understanding of institutional objects.
Institutional objects, such as money, drivers' licenses, and borders, have functions because of their social roles rather than their immediate physical properties. These objects are causally different than standard artifacts (e.g. hammers, chairs, and cars), sharing more commonality with other social roles. Thus, they inform psychological theories of human-made objects as well as children's emerging understanding of social reality. We examined whether children (N=180, ages 4-9) differentiate institutional objects from standard artifacts. Specifically, we examine whether children understand that mutual intentions (i.e., the intentions of a social collective) underlie the functional affordances of institutional objects in ways that they do not for standard artifacts. We find that young children assimilate institutional objects into their intuitive theories of standard artifacts; children begin to differentiate between the domains in the elementary school years.


Epitope mapping of Acinetobacter baumannii outer membrane protein W (OmpW) and laboratory study of an OmpW-derivative peptide.
Outer membrane protein W (OmpW) is a less-known A. baumannii antigen with potential immunogenic properties. The epitopes of this protein are not well-identified yet. Therefore, in the present study, B- and T-cell epitopes of A. baumannii OmpW were found using comprehensive in silico and partially in vitro studies. The T-cell (both class-I and class-II) and B-cell (both linear and conformational) epitopes were predicted and screened through many bioinformatics approaches including the prediction of IFN-γ production, immunogenicity, toxicity, allergenicity, human similarity, and clustering. A single 15-mer epitopic peptide containing a linear B-cell and both classes of T-cell epitopes were found and used for further assays. For in vitro assays, patient- and healthy control-derived peripheral blood mononuclear cells were stimulated with the 15-mer peptide, Phytohemagglutinin, or medium alone, and cell proliferation and IFN-γ production assays were performed. The bioinformatics studies led to mapping OmpW epitopes and introducing a 15-mer peptide. In vitro assays to some extent showed its potency in cell proliferation but not in IFN-γ induction, although the responses were not very expressive and faced some questions/limitations. In general, in the current study, we mapped the most immunogenic epitopes of OmpW that may be used for future studies and also assayed one of these epitopes in vitro, which was shown to have an immunogenicity potential. However, the induced immune responses were not strong which suggests that the present peptide needs a series of biotechnological manipulations to be used as a potential vaccine candidate. More studies in this field are recommended.


Antinuclear antibody positive autoimmune disorders in North India: an appraisal.
The autoimmune disorders (AID) have since long been considered to be commoner in Western world as compared to Asian countries. This, however, may not be true as in developing countries, there are incomplete epidemiological data and lack of advanced diagnostic facilities leading to under diagnosis in many cases. In this study, we performed an 11-year retrospective analysis of medical records of all clinically suspected and immunofluorescence antinuclear antibody test (IF-ANA)-positive cases. The IF-ANA-positive cases in the year 2006-2007 were further analyzed to find out the morbidity contribution by IF-ANA-positive AID. A total of 36,310 serum samples were screened for antinuclear antibody (ANA) between the years 1996 and 2006. The mean positivity was 12.3%. A constant and statistically significant increase in AID was noticed over the last 11 years. In the year 2006-2007, out of 3,435 suspected AID cases, 18.9% were ANA positive. Of these, 86.0% were adult patients with age ranging from 2¼ to 88 years. A female preponderance was also noted with a female-to-male ratio of 3:1. Among the ANA-positive connective tissue disorders (CTD), systemic lupus erythematosus was the most common clinical diagnosis (4.6/10,000 cases) followed by scleroderma (1.2/10,000) and overlap syndrome (0.7/10,000). Rheumatic, renal and hematopoietic systems were commonly involved. The overall frequency of CTD was 21%. The report is the first and largest hospital-based study from India, highlighting the rising incidence and clinical profile of ANA-positive AID.


Preparation of a Ni3Sn2 alloy-type anode embedded in carbon nanofibers by electrospinning for lithium-ion batteries.
A pure-phase Ni3Sn2 intermetallic alloy encapsulated in a carbon nanofiber matrix (Ni3Sn2@CNF) was successfully prepared by electrospinning and applied as anode for lithium-ion batteries. The physical and electrochemical properties of the Ni3Sn2@CNF were compared to that of pure CNF. The resultant Ni3Sn2@CNF anode produced a high initial discharge capacity of ∼1300 mA h g-1, later stabilizing and retaining ∼350 mA h g-1 (vs. 133 mA h g-1 for CNF) after 100 cycles at 0.1C. Furthermore, even at a high current density of 1C, it delivered a high initial discharge capacity of ∼1000 mA h g-1, retaining ∼313 mA h g-1 (vs. 66 mA h g-1 for CNF) at the 200th cycle. The superior electrochemical properties of the Ni3Sn2@CNF over CNF were attributed to the presence of electrochemically active Sn and decreased charge-transfer resistance with the alloy encapsulation, as confirmed from cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) results. Finally, post-mortem field-emission scanning electron microscopy (FE-SEM) images proved the preservation of the carbon nanofibers and the alloy after cycling, confirming the successful accommodation of the volume changes during the alloying/dealloying reactions of Sn in the Ni3Sn2@CNF.


A systematic review and meta-analysis of the effects of foam rolling on range of motion, recovery and markers of athletic performance.
To conduct a systematic review with meta-analysis assessing the effects of foam rolling on range of motion, laboratory- and field-based athletic measures, and on recovery. MEDLINE, PubMed, EMBASE, SPORTDiscus and Science Direct were searched (2005-June 2018). Experimental and observational studies were included if they examined the effects of foam rolling on measures of athletic performance in field or laboratory settings. Two investigators independently assessed methodologic quality using the Physiotherapy Evidence Database (PEDro) Scale. Study characteristics including participant age, sex and physical activity status, foam rolling protocol and pre- and post-intervention mean outcome measures were extracted. A total of 32 studies (mean PEDro = 5.56) were included in the qualitative analysis, which was themed by range of motion, laboratory-based measures, field-based measures and recovery. Thirteen range of motion studies providing 18 datasets were included in the meta-analysis. A large effect (d = 0.76, 95% CI 0.55-0.98) was observed, with foam rolling increasing range of motion in all studies in the analysis. Foam rolling increases range of motion, appears to be useful for recovery from exercise induced muscle damage, and there appear to be no detrimental effect of foam rolling on other athletic performance measures. However, except range of motion, it cannot be concluded that foam rolling is directly beneficial to athletic performance. Foam rolling does not appear to cause harm and seems to elicit equivalent effects in males and females.


Visible absorption bands of the (O2)2 collision complex at pressures below 760 Torr.
The collision-induced absorption of oxygen in the 540-650-nm wavelength region has been measured ata pressure range from 0 to 730 Torr at T 5 294 K. Pressure-dependent cross sections of the X 3Sigmag(+)+X 3Sigmag alpha 1Deltag(nu =0) + alpha 1Deltag(nu = 1 ) and X 3Simag(+)+ X 3Sigmag+--> alpha 1Deltag(nu =0) + 1 alpha 1Deltag (nu = 0) transitions have been determined by means of cavity-ringdown spectroscopy. Contributions of the overlapping g and d bands of O2 have been evaded, and Rayleigh extinction has been taken into account.


Assessment of the stereoselective fungal biotransformation of albendazole and its analysis by HPLC in polar organic mode.
A high-performance liquid chromatographic method using polar organic mode was developed to analyze albendazole (ABZ), albendazole sulfone (ABZSO(2)) and the chiral and active metabolite albendazole sulfoxide (ABZSOX, ricobendazole) that was further applied in stereoselective fungal biotransformation studies. The chromatographic separation was performed on a Chiralpak AS column using acetonitrile:ethanol (97:3, v/v) plus 0.2% triethylamine and 0.2% acetic acid as the mobile phase at a flow rate of 0.5 mL min(-1). The present study employed hollow fiber liquid-phase microextraction as sample preparation. The method showed to be linear over the concentration range of 25-5000 ng mL(-1) for each ABZSOX enantiomer, 200-10,000 ng mL(-1) for ABZ and 50-1000 ng mL(-1) for ABZSO(2) metabolite with correlation coefficient (r)>0.9934. The mean recoveries for ABZ, rac-ABZSOX and ABZSO(2) were, respectively, 9%, 33% and 20% with relative standard deviation below 10%. Within-day and between-day precision and accuracy assays for these analytes were studied at three concentration levels and were lower than 15%. This study opens the door regarding the possibility of using fungi in obtaining of the active metabolite ricobendazole. Nigrospora sphaerica (Sacc.) E. W. Mason (SS67), Pestalotiopsis foedans (VR8), Papulaspora immersa Hotson (SS13) and Mucor rouxii were able to stereoselectively metabolize ABZ into its chiral metabolite. Among them, the fungus Mucor rouxii was the most efficient in the production of (+)-ABZSOX.


Electronic properties and reactivity of oxidized graphene nanoribbons and their interaction with phenol.
The effect of the oxidized functional groups on the structural, electronic, and reactivity properties of armchair graphene nanoribbons has been investigated in the framework of the density functional theory. The presence of functional groups near the edges stabilizes the oxidized graphene nanoribbons (OGNRs) more than substituting near the center. Overall, we found slight differences in the electronic properties of OGNRs concerning the pristine ones. The oxygen contribution of functional groups to the DOS is found in the conducting energy bands far from the Fermi level. Consequently, the semiconducting behavior is maintained after doping. Based on the reactivity of OGNRs, the most promising nanostructures were proposed as adsorbents studying the interaction and complexation with phenol, a critical pollutant removed mainly by hydrotreating processes (HDO) to produce bio-oil. Parallel and perpendicular phenol conformations were found towards the OGNRs in the optimized complexes driven by a physisorption process. These results provide significant insights for catalytic processes that use biomass derivatives containing phenolic compounds. The physisorption of streams containing pollutants on OGNRs could be adapted to new technological applications for the remotion of aromatic compounds under environmentally friendly operational conditions.


Can developmental venous anomalies cause seizures?
Developmental venous anomalies (DVAs) are congenital anatomical variants of normal venous drainage of normal brain. Although DVAs are often discovered on the occasion of a seizure, their involvement in epilepsy is poorly studied. Our objective was to determine whether DVA can cause seizures, in the cases where there is no associated lesion, including no cavernoma or dysplasia. Based on clinical history, cerebral MRI, EEG recording, and 18F-FDG PET, we report 4 patients with DVA revealed by seizures. The first patient had a convulsive seizure caused by a hemorrhagic infarction due to thrombosis of her DVA. The second patient had a left temporo-parietal DVA next to a nonspecific lesion, possibly a sequelae of a venous infarction. The last two patients disclosed an isolated and uncomplicated DVA with a concordant epileptic focus confirmed on ictal video EEG recording. We reviewed literature and identified 21 other published cases of seizures caused by complications of a DVA and 9 patients that may have a direct link between epilepsy and an isolated and uncomplicated DVA. Seizures are linked to a DVA in two main situations: presence of an associated epileptogenic lesion, such as cavernoma or dysplasia, and occurrence of a complication of the DVA. Before concluding that a seizure is caused by a DVA, it is essential to perform full MRI protocols to search them. It remains rare and uncertain that isolated and uncomplicated DVA can cause seizures. In this last situation, physiopathological processes are probably different in each patient.


Reclassification of type 2 diabetes to type 1 diabetes in Asturias (Spain) between 2011 and 2020.
Differentiating between type 1 diabetes (T1D) and type 2 diabetes (T2D) can be difficult in adults. The aim of this study was to determine the frequency of diagnostic reclassification from T2D to T1D, the characteristics of the patients and the impact on the management of the disease. Observational and descriptive study including patients diagnosed with T1D in Asturias (Spain) between 2011 and 2020 who had been considered as T2D for at least 12 months. A total of 205 patients were included, representing 45.3% of those diagnosed with T1D over 30 years of age. Median time of evolution as T2D was 7,8 years. The age was 59.1 ± 12.9 years. BMI was > 25 kg/m2 in 46.8% of patients. HbA1c was 9.1 ± 2.1%, 77 ± 22 mmol/mol, and 56.5% were using insulin. Pancreatic antibodies were present in 95.5%, the most frequent being GAD, 82.6%. At 6 months, basal insulin use increased from 46.9 to 86.3%, and HbA1c decreased, 9.2 ± 2.0%vs7.7 ± 1.2%, 77 ± 22vs60 ± 13 mmol/mol; p < 0.0001. Diagnosis as T2D in patients with T1D in adults is common. Age, BMI, insulin use and other clinical features are not definitely discriminatory. GAD is the antibody of choice in case of diagnostic suspect. Reclassification has important implications for metabolic control.


Long term tolerability and clinical outcomes associated with tocilizumab in the treatment of refractory antibody mediated rejection (AMR) in pediatric renal transplant recipients.
Treatment options for antibody-mediated rejection (AMR) are limited. Recent studies have shown that inhibition of interleukin-6 (IL-6)/interleukin-6 receptor (IL-6R) signaling can reduce inflammation and slow AMR progression. We report our experience using monthly tocilizumab (anti-IL6R) in 25 pediatric renal transplant recipients with AMR, refractory to IVIg/Rituximab. From January 2013 to June 2019, a median (IQR) of 12 (6.019.0) doses of tocilizumab were given per patient. Serial assessments of renal function, biopsy findings, and HLA DSA (by immunodominant HLA DSA [iDSA] and relative intensity score [RIS]) were performed. Median (IQR) time from transplant to AMR was 41.4 (24.367.7) months, and time from AMR to first tocilizumab was 10.6 (8.317.6) months. At median (IQR) follow up of 15.8 (8.435.7) months post-tocilizumab initiation, renal function was stable except for 1 allograft loss. There was no significant decrease in iDSA or RIS. Follow up biopsies showed reduction in peritubular capillaritis (p = .015) and C4d scoring (p = .009). The most frequent adverse events were cytopenias. Tocilizumab in pediatric patients with refractory AMR was well tolerated and appeared to stabilize renal function. The utility of tocilizumab in the treatment of AMR in this population should be further explored.


Antenatal steroids and thyroid hormone function in preterm infants.
Antenatal steroids (ANS) are used widely for women at risk of preterm delivery. Evidence on the effects of ANS on thyroid hormone function in preterm infants is limited. To determine effects of ANS on thyroid hormone function in preterm infants. A retrospective cohort study of preterm infants born before 30 weeks of gestation. Infants were divided into no ANS, partial ANS, and complete ANS groups. Thyroid function tests at day of life 30 were compared. 260 Infants were included. A significantly higher proportion of patients were started on levothyroxine (LT4) in no ANS group and partial ANS group compared to complete ANS group. Logistic regression analysis revealed that infants in no ANS group are more likely to have TSH > 6 μIU ml-1 and started on LT4 compared to complete ANS group. Infants in no ANS group are more likely to have thyroid dysfunction compared to complete ANS group.


Structural and functional characterization of monomeric EphrinA1 binding site to EphA2 receptor.
The EphA2 receptor is overexpressed in glioblastoma multiforme and has been to shown to contribute to cell transformation, tumor initiation, progression, and maintenance. EphrinA1 (eA1) is a preferred ligand for the receptor. Treatment with monomeric eA1, the form of eA1 found in the extracellular environment, causes receptor phosphorylation, internalization, and down-regulation with subsequent anti-tumor effects. Here, we investigated the structure-function relationship of a monomeric eA1 focusing on its G-H loop ((108)FQRFTPFTLGKEFKE(123)G), a highly conserved region among eAs that mediates binding to their receptors. Alanine substitution mutants of the G-H loop amino acids were transfected into U-251 MG glioblastoma multiforme cells, and functional activity of each mutant in conditioned media was assessed by EphA2 down-regulation, ERK and AKT activation and cellular response assays. Alanine substitutions at positions Pro-113 Thr-115, Gly-117, Glu-122, and also Gln-109 enhanced the EphA2 receptor down-regulation and decreased p-ERK and p-AKT. Substitution mutants of eA1 at positions Phe-108, Arg-110, Phe-111, Thr-112, Phe-114, Leu-116, Lys-118, Glu-119, and Phe-120 had a deleterious effect on EphA2 down-regulation when compared with eA1-WT. Mutants at positions Phe-108, Lys-18, Lys-121, Gly-123 retained similar properties to eA1-WT. Recombinant eA1-R110A, -T115A, -G117A, and -F120A have been found to exhibit the same characteristics as the ligands contained in the conditioned media mainly due to the differences in their binding to the receptor. Thus, we have identified variants of eA1 that possess either superagonistic or antagonistic properties. These new findings will be important in the understanding of the receptor/ligand interactions and in further design of anti-cancer therapies targeting the eA/EphA system.


Chromosome-Scale Genome and Comparative Transcriptomic Analysis Reveal Transcriptional Regulators of β-Carotene Biosynthesis in Mango.
Mango (2n = 2x = 40) is an important tropical/subtropical evergreen fruit tree grown worldwide and yields nutritionally rich and high-value fruits. Here, a high-quality mango genome (396 Mb, contig N50 = 1.03 Mb) was assembled using the cultivar "Irwin" from Florida, USA. A total of 97.19% of the sequences were anchored to 20 chromosomes, including 36,756 protein-coding genes. We compared the β-carotene content, in two different cultivars ("Irwin" and "Baixiangya") and growth periods. The variation in β-carotene content mainly affected fruit flesh color. Additionally, transcriptome analysis identified genes related to β-carotene biosynthesis. MiPSY1 was proved to be a key gene regulating β-carotene biosynthesis. Weighted gene co-expression network analysis, dual luciferase, and yeast one-hybrid assays confirmed that transcription factors (TFs) MibZIP66 and MibHLH45 activate MiPSY1 transcription by directly binding to the CACGTG motif of the MiPSY1 promoter. However, the two TFs showed no significant synergistic effect on promoter activity. The results of the current study provide a genomic platform for studying the molecular basis of the flesh color of mango fruit.


Exopolysaccharide from Lactococcus hircilactis CH4 and Lactobacillus delbrueckii GRIPUMSK as new therapeutics to treat biofilm pathogens, oxidative stress and human colon adenocarcinoma.
Naturally occurring biopolymers like exopolysaccharides (EPS) secreted by lactic acid bacteria (LAB) has gained significant attention as they are cost effective, renewable and safe. In order to prevent the rapid increase in antibiotic resistant bacteria, the EPS of LAB offers novel approach of targeting the antibiotic resistant pathogens by limiting their effects on environment. Accordingly, in this study, the production, purification, characterization and biological properties of exopolysaccharides from Lactococcus hircilactis strain CH4 and Lactobacillus delbrueckii strain GRIPUMSK were performed. The optimization of lactic acid bacterial strains for exopolysaccharide production was done by response surface methodology and changing the carbon sources in the growth media. The carbohydrate and protein of exopolysaccharide 1 were 79.7 % and 8.7 % respectively and exopolysaccharide 2 were 75.2 % and 9.3 % respectively. When compared with the commercial emulsifier sodium dodecyl sulfate, both the exopolysaccharides have shown good emulsifying activity. Both the exopolysaccharides were linear homo-polysaccharide as determined by Fourier transform infrared spectroscopy and Nuclear magnetic resonance spectra. Scanning electron microscopy showed that the exopolysaccharides were porous and capable of holding water. The exopolysaccharides were partially crystalline as confirmed by X-ray diffraction spectra. Exopolysaccharides from L. hircilactis and L. delbrueckii exhibited significant antimicrobial activity against H. pylori, S. flexneri, S. pyogenes, E. faecalis and C. albicans. Both the exopolysaccharides revealed significant 2,2-diphenyl-1-picrylhydrazyl and hydrogen peroxide scavenging ability with the IC50 value of 100 μg/ml and 80 μg/ml respectively. Exopolysaccharides from L. hircilactis and L. delbrueckii at 100 μg/ml showed significant anticancer activity on HT-29 cells with 58.4 % and 58.7 % respectively. These findings proved that exopolysaccharides from the two selected lactic acid bacterial strains could be explored as natural bioactive carbohydrate polymer for biomedical applications.


Oxalate:formate exchange. The basis for energy coupling in Oxalobacter.
In the Gram-negative anaerobe, Oxalobacter formigenes, the generation of metabolic energy depends on the transport and decarboxylation of oxalate. We have now used assays of reconstitution to study the movements of oxalate and to characterize the exchange of oxalate with formate, its immediate metabolic derivative. Membranes of O. formigenes were solubilized with octyl-beta-D-glucopyranoside in the presence of 20% glycerol and Escherichia coli phospholipid, and detergent extracts were reconstituted by detergent dilution. [14C]Oxalate was taken up by proteoliposomes loaded with unlabeled oxalate, but not by similarly loaded liposomes or by proteoliposomes containing sulfate in place of oxalate. Oxalate transport did not depend on the presence of sodium or potassium, nor was it affected by valinomycin (1 microM), nigericin (1 microM), or a proton conductor, carbonylcyanide-p-trifluoromethoxyphenylhydrazone (5 microM) when potassium was at equal concentration on either side of the membrane. Such data suggest the presence of an overall neutral oxalate self-exchange, independent of common cations or anions. Kinetic analysis of the reaction in proteoliposomes gave a Michaelis constant (Kt) for oxalate transport of 0.24 mM and a maximal velocity (Vmax) of 99 mumol/min/mg of protein. A direct exchange of oxalate and formate was indicated by the observations that formate inhibited oxalate transport and that delayed addition of formate released [14C]oxalate accumulated during oxalate exchange. Moreover, [14C]formate was taken up by oxalate-loaded proteoliposomes (but not liposomes), and this heterologous reaction could be blocked by external oxalate. Further studies, using formate-loaded proteoliposomes, suggested that the heterologous exchange was electrogenic. Thus, for assays in which N-methylglucamine served as both internal and external cation, formate-loaded particles took up oxalate at a rate of 2.4 mumol/min/mg of protein. When external or internal N-methylglucamine was replaced by potassium in the presence of valinomycin, there was, respectively, a 7-fold stimulation or an 8-fold inhibition of oxalate accumulation, demonstrating that net negative charge moved in parallel with oxalate during the heterologous exchange. The work summarized here suggests the presence of an unusually rapid and electrogenic oxalate2-:formate1- antiport in membranes of O. formigenes. Since a proton is consumed during the intracellular decarboxylation that converts oxalate into formate plus CO2, antiport of oxalate and formate would play a central role in a biochemical cycle consisting of (a) oxalate influx, (b) oxalate decarboxylation, and (c) formate efflux.(ABSTRACT TRUNCATED AT 400 WORDS)


Disruption of dual homeostasis by a metal-organic framework nanoreactor for ferroptosis-based immunotherapy of tumor.
Ferroptosis, a newfound non-apoptotic cell death pathway that is iron- and reactive oxygen species (ROS)-dependent, has shown a promise for tumor treatment. However, engineering ferroptosis inducers with sufficient hydrogen peroxide (H2O2) and iron supplying capacity remains a great challenge. To address this issue, herein, we report a powerful nanoreactor by modifying MnO2, glucose oxidase, and polyethylene glycol on iron-based metal-organic framework nanoparticles for disrupting redox and iron metabolism homeostasis, directly providing the Fenton reaction-independent downstream ferroptosis for tumor therapy. By consuming glutathione and oxidizing glucose to increase the H2O2 level in cancer cells and downregulating ferroportin 1 to accumulate intracellular iron ions, the homeostasis disruptor could effectively enhance the ferroptosis. Subsequently, the ferroptosis cells release tumor immune-associated antigens, which combine with in situ injected aptamer-PD-L1 to further strengthen the tumor treatment efficiency. This work not only paves a way to enhance the efficacy of ferroptosis-based cancer therapy by associating intracellular redox homeostasis with the iron metabolism system in tumor cells but also offers an engineered nanoreactor as a promising mimetic antigen for activating immunotherapy.


To Be or Not to Be Patent: The Relationship Between Migraine and Patent Foramen Ovale.
Migraine is a common neurological disorder with an uncertain pathogenesis. A pathophysiological link has been proposed between the migraine headache and patent foramen ovale (PFO). However, the data about the association of migraine with the presence of PFO are conflicting. The study aims to prospectively investigate the prevalence of PFO in patients with migraine compared with that of control subjects. A total of 203 migraineous patients and 212 control subjects with similar demographic characteristics to that of case subjects regarding age and gender were included in the study. Transthoracic echocardiography with agitated saline injection was used to evaluate the presence of PFO. PFO was judged to be present if any microbubble was seen in the left cardiac chambers within the first 5 cardiac cycles from the maximum right atrial opacification at rest or after provocative maneuvers. Mean age of the patients was 37.2 ± 9.8 years and 93% were female. Migraine with aura was present in 32% (n = 65) of the patients. The prevalence of PFO was similar in patients with migraine and the control subjects (42% vs 44%; odds ratio 0.90, 95% confidence interval 0.61-1.33, P = .61). Likewise, the prevalence of PFO was similar in migraineurs patients with or without aura (41% vs 42%; odds ratio 0.95, 95% confidence interval 0.51-1.76, P = .87). This study suggest that PFO is not more common in patients with migraine compared with healthy subjects. Therefore, our findings suggest that the relationship between migraine headache and the presence of PFO is questionable.


Anti-allergic substances contained in the pollen of Cryptomeria japonica possess diverse effects on the degranulation of RBL-2H3 cells.
Following prolonged exposure to some of the flavonoids with RBL-2H3 cells, secretion of hexosaminidase, a granule constituent, stimulated by an immunologic was enhanced. RBL-2H3 cells do not normally respond to polybasic secretagogues, but as reported here, they do so after prolonged exposure. Effect of flavonoids on secretion of hexosaminidase was also investigated. Of the thirteen flavonoids, quercetin and fisetin were the most potent inhibitors. A structure-activity study indicated that the position, number, and substitution of the hydroxy group of the B ring and saturation of the C2-C3 bond are important factors affecting flavonoid inhibition of secretary granules in RBL-2H3 cells.


Development and Optimization of Clinical Informatics Infrastructure to Support Bioinformatics at an Oncology Center.
Translational bioinformatics for therapeutic discovery requires the infrastructure of clinical informatics. In this chapter, we describe the clinical informatics components needed for successful implementation of translational research at a cancer center. This chapter is meant to be an introduction to those clinical informatics concepts that are needed for translational research. For a detailed account of clinical informatics, the authors will guide the reader to comprehensive resources. We provide examples of workflows from Moffitt Cancer Center led by Drs. Perkins and Markowitz. This perspective represents an interesting collaboration as Dr. Perkins is the Chief Medical Information Officer and Dr. Markowitz is a translational researcher in Melanoma with an active informatics component to his laboratory to study the mechanisms of resistance to checkpoint blockade and an active member of the clinical informatics team.


A review of randomized trials comparisons of epidural with parenteral forms of pain relief during labour and its impact on operative and cesarean delivery rate.
The aim of this review was to summarise the available literature on different modalities of labour pain relief (epidural vs. parenteral) and to assess their impact on the rate of caesarean section deliveries and instrumental deliveries, and on the need to support the uterine contractile function. The PubMed, Web of Science and Cochrane databases were reviewed to identify articles describing the effect of labour pain relief on the course of labour. This review includes 16 studies with 7150 patients. The analysis of the obtained data revealed that epidural analgesia (EA) or combined epidural and spinal anaesthesia (CESA) provided significantly better labour pain relief when compared with parenteral opioids. Conduction anaesthesia was not associated with an increase in the caesarean section delivery rate. Some authors concluded that conduction anaesthesia was associated with the need for assisted delivery. Epidural analgesia is a well-recognised method of labour pain relief. It is associated with the parturient's higher satisfaction when compared to parenteral opioids. EA does not directly increase the caesarean section delivery rate, yet it can lead to instrumental deliveries (vacuum-assisted, obstetrics forceps) and a need to pharmacologically support the uterine contractile function. Further studies are required to evaluate the effect of EA on the course of labour, and methods of minimising its adverse effects.