Advantages of bioconjugated silica-coated nanoparticles as an innovative diagnosis for human toxoplasmosis.
Nanotechnology is a promising arena for generating new applications in Medicine. To successfully functionalised nanoparticles for a given biomedical application, a wide range of chemical, physical and biological factors have to be taken into account. Silica-coated nanoparticles, (SiO2NP) exhibit substantial diagnostic activity owing to their large surface to volume ratios and crystallographic surface structure. This work aimed to evaluate the advantage of bioconjugation of SiO2NP with PAb against Toxoplasma lyzate antigen (TLA) as an innovative diagnostic method for human toxoplasmosis. This cross-sectional study included 120 individuals, divided into Group I: 70 patients suspected for Toxoplasma gondii based on the presence of clinical manifestation. Group II: 30 patients harboring other parasites than T. gondii Group III: 20 apparently healthy individuals free from toxoplasmosis and other parasitic infections served as negative control. Detection of circulating Toxoplasma antigen was performed by Sandwich ELISA and Nano-sandwich ELISA on sera and pooled urine of human samples. Using Sandwich ELISA, 10 out of 70 suspected Toxoplasma-infected human serum samples showed false negative and 8 out of 30 of other parasites groups were false positive giving 85.7% sensitivity and 84.0% specificity, while the sensitivity and specificity were 78.6% and 70% respectively in urine samples. Using Nano-Sandwich ELISA, 7 out of 70 suspected Toxoplasma-infected human samples showed false negative results and the sensitivity of the assay was 90.0%, while 4 out of 30 of other parasites groups were false positive giving 92.0% specificity, while the sensitivity and specificity were 82.6% and 80% respectively in urine samples. In conclusion, our data demonstrated that loading SiO2 nanoparticles with pAb increased the sensitivity and specificity of Nano-sandwich ELISA for detection of T.gondii antigens in serum and urine samples, thus active (early) and light infections could be easily detected.


A Simple Methodology to Differentiate Changes in Bioavailability From Changes in Clearance Following Oral Dosing of Metabolized Drugs.
Accurately discriminating changes in clearance (CL) from changes in bioavailability (F) following an oral drug-drug interaction is difficult without carrying out an intravenous interaction study. This may be true for drugs that are clinically significant transporter substrates; however, for interactions that are strictly metabolic, it has been recognized that volume of distribution remains unchanged between both phases of the interaction study. With the understanding that changes in volume of distribution will be minimal for metabolized drugs, the inverse of the change in apparent volume of distribution can provide adequate estimates of the change in bioavailability alone. Utilization of this estimate of F change in tandem with the observed apparent clearance (CL/F) change in an oral drug-drug interaction can provide an estimate of the change in clearance alone. Here, we examine drug-drug interactions involving five known inhibitors and inducers of cytochrome P450 3A4 isozyme on victim drugs midazolam and apixaban for which the interaction was carried out both orally and intravenously, allowing for evaluation of this methodology. Predictions of CL and F changes based on oral data were reasonably close to observed changes based on intravenous studies, demonstrating that this simple yet powerful methodology can reasonably differentiate changes in F from changes in CL for oral metabolic drug interactions when only oral data are available. Utilization of this relatively simple methodology to evaluate DDIs for orally dosed drugs will have a significant impact on how DDIs are interpreted from a drug development and regulatory perspective.


Solid phase synthesis of the protected 43-55 tridecapeptide of the heavy chain of myeloma immunoglobin M603, employing cyclohexyl ester protection for glutamic acid.
A protected tridecapeptide of the sequence Boc-Lys(2CIZ)-Arg(Tos)-Leu-Glu (OcHex)-Trp(For)-Ile-Ala-Ala-Ser(Bzl)-Arg(Tos)-Asn-Lys(2CIZ)-Gly-OH, representing residues 43-55 of the variable region of the heavy chain of mouse myeloma protein M603, was synthesized. It was assembled by a stepwise solid phase method designed to give a fully protected peptide in high yield and purity with minimal side reactions. Thus, the peptide chain was attached as an alpha-methyl phenacyl ester to a 2-bromopropionyl-resin. After the synthesis the protected peptide fragment was obtained in 89% yield by photolytic cleavage from the resin. The peptide was purified by multiple precipitation and column chromatography. It was shown to be homogeneous by reverse phase high pressure liquid chromatography, and it had the correct amino acid composition and sequence. In the course of this work it was shown that tert.-butyloxycarbonyl-amino acids caused the formation of significant amounts of pyrrolidone carboxylic acid residues during the coupling reaction when a gamma-benzyl glutamyl residue was NH2-terminal. Other weak-acid additives also caused this chain terminating side reaction. The cyclization was markedly suppressed by protection of the glutamyl side chain as a cyclohexyl ester. With this protecting group, no evidence of pyrrolidone carboxylic acid formation could be detected in the tridecapeptide 43-55.


A host protein (La) binds to a unique species of minus-sense leader RNA during replication of vesicular stomatitis virus.
Baby hamster kidney cells infected with the minus-strand RNA virus vesicular stomatitis virus (VSV) were found to contain three small viral leader RNA species of the minus sense. The longest minus-strand leader RNA was 54 nucleotides long and was complexed with the host cell La protein that was immunoprecipitated by antisera from patients with systemic lupus erythematosus. The La protein is normally found associated with RNA polymerase III transcripts in their unprocessed form. Shorter minus-strand leader RNA species of 45-48 nucleotides were more abundant but were not associated with the La protein. Unlike the plus-strand leader RNA of VSV, the minus-strand leader RNAs were not detected in the nucleus in any form. The minus-strand leader RNAs accumulated gradually throughout the infection and could not be found in association with the viral nucleocapsid protein. The sequence required for La protein binding on the 54-nucleotide-long minus-strand leader is similar to that at the 3' end of the La protein binding-plus-strand leader RNA and, thus, we propose a role for the La protein in the replication of VSV.


The receptor-mediated activation of tyrosine hydroxylation in the superior cervical ganglion of the rat.
The addition of carbachol to superior cervical ganglia causes a rapid increase in tyrosine hydroxylation in situ. The increase occurs in ganglia from both newborn and adult animals, and in ganglia from animals pretreated with reserpine. The increase is not due to increased transport of the substrate. The increase is dependent upon the presence of calcium, and is additive to the stimulation produced by dibutyryl cyclic AMP. The stimulation seems specific for tyrosine hydroxylation; dopamine beta-hydroxylation is not increased. Preincubation experiments suggest that the carbachol-induced stimulation is due to a change in the availability of, or the affinity of the enzyme for, reduced pterin cofactor. The stimulation is inhibited by atropine and also by low concentrations of phenoxybenzamine or haloperidol, which suggests that it is caused by an action of carbachol on the interneurons in the ganglia.


Balteatide: a novel antimicrobial decapeptide from the skin secretion of the purple-sided leaf frog, Phyllomedusa baltea.
The skin secretions of Neotropical phyllomedusine leaf frogs have proven to be a rich source of biologically active peptides, including antimicrobials. The major families of antimicrobial peptides (AMPs) reported are the dermaseptins and phylloseptins and the minor families are the dermatoxins, phylloxins, plasticins, distinctins, and medusins. Here, we report a novel AMP of 10 amino acid residues (LRPAILVRIKamide), named balteatide, from the skin secretion of wild Peruvian purple-sided leaf frogs, Phyllomedusa baltea. Balteatide was found to exhibit a 90% sequence identity with sauvatide, a potent myotropic peptide from the skin secretion of Phyllomedusa sauvagei. However, despite both peptides exhibiting only a single amino acid difference (I/T at position 9), sauvatide is devoid of antimicrobial activity and balteatide is devoid of myotropic activity. Balteatide was found to have differential activity against the Gram-positive bacterium, Staphylococcus aureus; the Gram-negative bacterium, Escherichia coli; and the yeast, Candida albicans, and unusual for phyllomedusine frog skin AMPs, was most potent (MIC 32 mg/L) against the yeast. Balteatide was also devoid of haemolytic activity up to concentrations of 512 mg/L. Phyllomedusine frog skin secretions thus continue to provide novel AMPs, some of which may provide templates for the rational design of new classes of anti-infective therapeutics.


Genetically Diverse Low Pathogenicity Avian Influenza A Virus Subtypes Co-Circulate among Poultry in Bangladesh.
Influenza virus surveillance, poultry outbreak investigations and genomic sequencing were assessed to understand the ecology and evolution of low pathogenicity avian influenza (LPAI) A viruses in Bangladesh from 2007 to 2013. We analyzed 506 avian specimens collected from poultry in live bird markets and backyard flocks to identify influenza A viruses. Virus isolation-positive specimens (n = 50) were subtyped and their coding-complete genomes were sequenced. The most frequently identified subtypes among LPAI isolates were H9N2, H11N3, H4N6, and H1N1. Less frequently detected subtypes included H1N3, H2N4, H3N2, H3N6, H3N8, H4N2, H5N2, H6N1, H6N7, and H7N9. Gene sequences were compared to publicly available sequences using phylogenetic inference approaches. Among the 14 subtypes identified, the majority of viral gene segments were most closely related to poultry or wild bird viruses commonly found in Southeast Asia, Europe, and/or northern Africa. LPAI subtypes were distributed over several geographic locations in Bangladesh, and surface and internal protein gene segments clustered phylogenetically with a diverse number of viral subtypes suggesting extensive reassortment among these LPAI viruses. H9N2 subtype viruses differed from other LPAI subtypes because genes from these viruses consistently clustered together, indicating this subtype is enzootic in Bangladesh. The H9N2 strains identified in Bangladesh were phylogenetically and antigenically related to previous human-derived H9N2 viruses detected in Bangladesh representing a potential source for human infection. In contrast, the circulating LPAI H5N2 and H7N9 viruses were both phylogenetically and antigenically unrelated to H5 viruses identified previously in humans in Bangladesh and H7N9 strains isolated from humans in China. In Bangladesh, domestic poultry sold in live bird markets carried a wide range of LPAI virus subtypes and a high diversity of genotypes. These findings, combined with the seven year timeframe of sampling, indicate a continuous circulation of these viruses in the country.


Gingival Exudatome Dynamics Implicate Inhibition of the Alternative Complement Pathway in the Protective Action of the C3 Inhibitor Cp40 in Nonhuman Primate Periodontitis.
Periodontitis is a prevalent chronic inflammatory disease associated with dysbiosis. Although complement inhibition has been successfully used to treat periodontitis in animal models, studies globally analyzing inflamed tissue proteins to glean insight into possible mechanisms of action are missing. Using quantitative shotgun proteomics, we aimed to investigate differences in composition of inflammatory gingival tissue exudate ("gingival crevicular fluid"; GCF), before and after local administration of an inhibitor of the central complement component, C3, in nonhuman primates. The C3 inhibitor, Cp40 (also known as AMY-101) was administered locally in the maxillary gingival tissue of cynomolgus monkeys with established periodontitis, either once a week (1×-treatment; n = 5 animals) or three times per week (3×-treatment; n = 10 animals), for 6 weeks followed by another 6 weeks of observation in the absence of treatment. 45 GCF samples were processed for FASP digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Data were processed using the ProgenesisQI software. The statistical significance of differences between the groups was determined by RM-ANOVA, and a protein expression change was considered as a true regulation at >2-fold and p < 0.05. The human orthologues were subjected to Gene Ontology analyses using PANTHER. Data are available via ProteomeXchange with identifier PXD009502. 573 proteins with >2 peptides were longitudinally quantified. Both 3× and 1× administration of Cp40 resulted in significant down-regulation of dozens of proteins during the 6-week course of treatment as compared to baseline. Following drug withdrawal at 6 weeks, more than 50% of the down-regulated proteins showed increased levels at week 12. The top scored pathway was "complement activation, alternative pathway", and several proteins involved in this pathway were down-regulated at 6 weeks. We mapped the proteomic fingerprint changes in local tissue exudate of cynomolgus monkey periodontitis in response to C3 inhibition and identified the alternative pathway of complement activation and leukocyte degranulation as main targets, which are thus likely to play significant roles in periodontal disease pathogenesis. Label-free quantitative proteomics strategies utilizing GCF are powerful tools for the identification of treatment targets and providing insights into disease mechanisms.


Building communities in times of crisis - Impacts of the COVID-19 pandemic on the work of transition intermediaries in the energy sector.
The ongoing COVID-19 pandemic affects people worldwide. The policies in response to the virus range from closure of national borders to curfews for entire metropolises, like Paris. While we can expect severe impacts on the world economy, the consequences of the pandemic for local sustainability transitions are entirely unclear. In this exploratory study, we investigate how the current situation affects the work of transition intermediaries in the energy sector. More concretely, we aim to analyse the impact of COVID-19 policies on community energy projects and the subsequent change of work practices of intermediaries in this situation. Our data consists of qualitative data we collected between January and October 2020. Our results show that transition intermediaries are affected in different ways. Most notably, the work on networking suffers during these times of crisis. We found that intermediaries are particularly challenged in their ability to build trust. This particularly affects new and complex community energy projects and intermediation activities needed for systemic change. We found that established projects with a strong trust base are least affected by these limitations. Intermediaries dependent on private funding face much bigger problems than publically funded organisations. Our results offer some novel and relevant insights in the role and work of transition intermediaries and the development of community energy projects in times of crisis. These findings can help governments, intermediary organizations and citizen groups to design future transition processes in ways that are more resilient to external shocks.


The feasibility of replacement therapy for inherited disorder of glycolysis: triosephosphate isomerase deficiency (review).
Triosephosphate isomerase (TPI, EC 126.96.36.199) is an ubiquitously expressed enzyme that catalyses the interconversion of dihydroxyacetone phosphate (DHAP) and glyceraldehyde-3-phosphate in the energy-generating glycolytic pathway. Inherited defects in the TPI gene are characterised biochemically by markedly reduced TPI enzyme activity in all tissues resulting in metabolic block in glycolysis, with accumulating DHAP particularly in red cells. Clinical TPI deficiency is a rare autosomal recessive multi-system disorder characterised by non-spherocytic haemolytic anaemia, recurrent infections, cardiomyopathy, severe and fatal neuromuscular dysfunctions. Reviews of current literature show that after 30 years since TPI deficiency was first described, the disease still remains without effective therapy. However, several potential therapeutic strategies exist for the treatment of inherited metabolic disorders such as TPI deficiency. Development of an effective therapy for TPI deficiency presents a fascinating and formidable challenge for basic laboratory and clinical research. The major aim of this overview is to discuss the current knowledge of TPI deficiency with special emphasis on research efforts directed towards reversing the metabolic effects of the disorder.


Relationship between Quality of Life and Adult Attachment Factors in Mothers of Children with and without Type 1 Diabetes.
The mothers of children with a specific clinical situation such as type 1 diabetes mellitus may have a higher level of stress, causing a worse perception of their quality of life, greater anxiety, and greater avoidance (adult attachment factors). The objective of this research was to verify if there is a relationship between the adult attachment factors of mothers of children with and without type 1 diabetes mellitus and the perception of the quality of life of these mothers. This survey was carried out from July to September 2022, with mothers of children aged 5 to 10 years, with and without diabetes. The data were collected through an online questionnaire, with socioeconomic data from the attachment scale Experience in Close Relationship-(Reduced), and the questionnaire on the quality of life, the WHOQOL-abbreviated questionnaire. For statistical analysis, the Statistical Package for Social Science 24 was used. A total of 45 mothers of children with DM1 and 55 mothers of healthy children were evaluated. The mothers of children with DM1 had a worse perception of their quality of life when compared to the mothers of healthy children (p < 0.05), with no difference in terms of the attachment style. Therefore, it is understood that actions aimed at improving the quality of life of these mothers are necessary.


A randomized, placebo-controlled trial of corticosteroids for hyperemesis due to pregnancy.
Hyperemesis gravidarum, a severe form of nausea and vomiting due to pregnancy for which there is no proven pharmacological treatment, is the third leading cause for hospitalization during pregnancy. Corticosteroids are commonly used for the treatment of nausea and vomiting due to cancer chemotherapy-induced emesis and might prove useful in hyperemesis gravidarum. A randomized, double-blind, placebo-controlled trial was conducted in 126 women who previously had not responded to outpatient therapy for hyperemesis gravidarum during the first half of pregnancy. Intravenous methylprednisolone (125 mg) was followed by an oral prednisone taper (40 mg for 1 day, 20 mg for 3 days, 10 mg for 3 days, 5 mg for 7 days) versus an identical-appearing placebo regimen. All women also received promethazine 25 mg and metoclopramide 10 mg intravenously every 6 hours for 24 hours, followed by the same regimen administered orally as needed until discharge. The primary study outcome was the number of women requiring rehospitalization for hyperemesis gravidarum. A total of 110 women delivered at our hospital and had pregnancy outcomes available for analysis; 56 were randomized to corticosteroids and 54 were administered placebo. Nineteen women in each study group required rehospitalization (34% versus 35%, P =.89, for corticosteroids versus placebo, respectively). The addition of parenteral and oral corticosteroids to the treatment of women with hyperemesis gravidarum did not reduce the need for rehospitalization later in pregnancy.


Comparison of Swine and Human Computational Hemodynamics Models for the Study of Coronary Atherosclerosis.
Coronary atherosclerosis is a leading cause of illness and death in Western World and its mechanisms are still non completely understood. Several animal models have been used to 1) study coronary atherosclerosis natural history and 2) propose predictive tools for this disease, that is asymptomatic for a long time, aiming for a direct translation of their findings to human coronary arteries. Among them, swine models are largely used due to the observed anatomical and pathophysiological similarities to humans. However, a direct comparison between swine and human models in terms of coronary hemodynamics, known to influence atherosclerotic onset/development, is still lacking. In this context, we performed a detailed comparative analysis between swine- and human-specific computational hemodynamic models of coronary arteries. The analysis involved several near-wall and intravascular flow descriptors, previously emerged as markers of coronary atherosclerosis initiation/progression, as well as anatomical features. To do that, non-culprit coronary arteries (18 right-RCA, 18 left anterior descending-LAD, 13 left circumflex-LCX coronary artery) from patients presenting with acute coronary syndrome were imaged by intravascular ultrasound and coronary computed tomography angiography. Similarly, the three main coronary arteries of ten adult mini-pigs were also imaged (10 RCA, 10 LAD, 10 LCX). The geometries of the imaged coronary arteries were reconstructed (49 human, 30 swine), and computational fluid dynamic simulations were performed by imposing individualized boundary conditions. Overall, no relevant differences in 1) wall shear stress-based quantities, 2) intravascular hemodynamics (in terms of helical flow features), and 3) anatomical features emerged between human- and swine-specific models. The findings of this study strongly support the use of swine-specific computational models to study and characterize the hemodynamic features linked to coronary atherosclerosis, sustaining the reliability of their translation to human vascular disease.


Phospholipase D regulates myogenic differentiation through the activation of both mTORC1 and mTORC2 complexes.
How phospholipase D (PLD) is involved in myogenesis remains unclear. At the onset of myogenic differentiation of L6 cells induced by the PLD agonist vasopressin in the absence of serum, mTORC1 complex was rapidly activated, as reflected by phosphorylation of S6 kinase1 (S6K1). Both the long (p85) and short (p70) S6K1 isoforms were phosphorylated in a PLD1-dependent way. Short rapamycin treatment specifically inhibiting mTORC1 suppressed p70 but not p85 phosphorylation, suggesting that p85 might be directly activated by phosphatidic acid. Vasopressin stimulation also induced phosphorylation of Akt on Ser-473 through PLD1-dependent activation of mTORC2 complex. In this model of myogenesis, mTORC2 had a positive role mostly unrelated to Akt activation, whereas mTORC1 had a negative role, associated with S6K1-induced Rictor phosphorylation. The PLD requirement for differentiation can thus be attributed to its ability to trigger via mTORC2 activation the phosphorylation of an effector that could be PKCα. Moreover, PLD is involved in a counter-regulation loop expected to limit the response. This study thus brings new insights in the intricate way PLD and mTOR cooperate to control myogenesis.


D1 receptor in interneurons of macaque prefrontal cortex: distribution and subcellular localization.
Working memory performance is influenced by dopamine activation of D1 family dopamine receptors in the prefrontal cortex; working memory performance is maximal at moderate stimulation of D1 family receptors and is reduced by either higher or lower levels of D1 stimulation. The neuronal mechanisms that underlie this complex relationship are not yet understood. Previous work from this laboratory has demonstrated that the D1 family receptors, D1 and D5, are located in different compartments of pyramidal cells. Here we use an antibody specific to the D1 receptor and double-label immunohistochemistry at the light and electron microscopic level to demonstrate that D1-like immunoreactivity (D1-LIR) is also present in interneurons. D1 receptor is prevalent in parvalbumin-containing interneurons and is less common in calretinin-containing interneurons. At the ultrastructural level, D1-LIR is found associated with the Golgi apparatus and endoplasmic reticulum in the soma, with the membranes of vesicles in proximal dendrites, and with the plasma membrane on distal dendrites, where it is often located near asymmetric synapses. In addition, D1-LIR is also seen in presynaptic axon terminals, which give rise to symmetric synapses onto dendritic shafts and soma. These results raise the possibility that the circuit basis of working memory in the prefrontal cortex involves a D1-mediated inhibitory component.


Meta-analysis of genome-wide association studies for height and body mass index in ∼700000 individuals of European ancestry.
Recent genome-wide association studies (GWAS) of height and body mass index (BMI) in ∼250000 European participants have led to the discovery of ∼700 and ∼100 nearly independent single nucleotide polymorphisms (SNPs) associated with these traits, respectively. Here we combine summary statistics from those two studies with GWAS of height and BMI performed in ∼450000 UK Biobank participants of European ancestry. Overall, our combined GWAS meta-analysis reaches N ∼700000 individuals and substantially increases the number of GWAS signals associated with these traits. We identified 3290 and 941 near-independent SNPs associated with height and BMI, respectively (at a revised genome-wide significance threshold of P < 1 × 10-8), including 1185 height-associated SNPs and 751 BMI-associated SNPs located within loci not previously identified by these two GWAS. The near-independent genome-wide significant SNPs explain ∼24.6% of the variance of height and ∼6.0% of the variance of BMI in an independent sample from the Health and Retirement Study (HRS). Correlations between polygenic scores based upon these SNPs with actual height and BMI in HRS participants were ∼0.44 and ∼0.22, respectively. From analyses of integrating GWAS and expression quantitative trait loci (eQTL) data by summary-data-based Mendelian randomization, we identified an enrichment of eQTLs among lead height and BMI signals, prioritizing 610 and 138 genes, respectively. Our study demonstrates that, as previously predicted, increasing GWAS sample sizes continues to deliver, by the discovery of new loci, increasing prediction accuracy and providing additional data to achieve deeper insight into complex trait biology. All summary statistics are made available for follow-up studies.


Transcriptional Response of Vitellogenin Gene in Flatfish to Environmental Pollutants from Two Regions of the Gulf of Mexico.
The present study evaluates the endocrine effect in flatfish through vitellogenin (vtg) gene expression and its association with pollutants data obtained from fish muscle and sediment from two regions in the Gulf of Mexico (GoM): Perdido Fold Belt (northwestern) and the Yucatan Peninsula (southeast). The results revealed induction of vtg in male flatfish in both geographical regions with different levels and patterns of distribution per oceanographic campaign (OC). In the Perdido Fold Belt, vtg was observed in male fish during four OC (carried out in 2016 and 2017), positively associated with Pb, V, Cd and bile metabolites (hydroxynaphthalene and hydroxyphenanthrene). In the Yucatan Peninsula, the induction of vtg in males was also detected in three OC (carried out in 2016 and 2018) mainly associated with Ni, Pb, Al, Cd, V and polycyclic aromatic hydrocarbons. Ultimately, estrogenic alterations could affect reproductive capacity of male flatfish in the GoM.


A prototype instrument for single pinhole small animal adaptive SPECT imaging.
The authors have designed and constructed a small-animal adaptive SPECT imaging system as a prototype for quantifying the potential benefit of adaptive SPECT imaging over the traditional fixed geometry approach. The optical design of the system is based on filling the detector with the region of interest for each viewing angle, maximizing the sensitivity, and optimizing the resolution in the projection images. Additional feedback rules for determining the optimal geometry of the system can be easily added to the existing control software. Preliminary data have been taken of a phantom with a small, hot, offset lesion in a flat background in both adaptive and fixed geometry modes. Comparison of the predicted system behavior with the actual system behavior is presented, along with recommendations for system improvements.


Steroid Use Associated With Increased Odds of 30-Day Mortality in Surgical Patients With Metastatic Spinal Tumors in the Setting of Disseminated Disease.
Steroid administration is part of a standard treatment regimen in metastatic spinal cord compression, though the appropriate dose, duration, efficacy, and risks remain controversial. To analyze the risk of preoperative steroid use on 30-d mortality in surgical metastatic spinal tumors with dissemination disease using a large multicenter national database. Adult patients who underwent surgical treatment for metastatic spine tumors between 2005 and 2014 were identified in the American College of Surgeons National Surgical Quality Improvement Program database. Demographic, preoperative risk factors, operative information, and postoperative events were extracted. Multivariate logistical regression modeling was used to investigate the association with preoperative steroid use with the outcome of interest, 30-d mortality. Other independent risk factors associated with 30-d mortality were also identified. Five hundred fifty-two patients underwent surgical treatment of spinal metastases with disseminated cancer present at time of surgery. Independent risk factors of 30-d mortality included prolonged steroid use (odds ratio [OR] 2.48, 95% confidence interval [CI]: 1.22-5.04, P = .012), dependent functional status (OR 2.91, 95% CI: 1.68-5.04, P < .001), history of bleeding disorder (OR 2.80, 95% CI: 1.16-6.74, P = .021), history of smoking (OR 2.26, 95% CI: 1.11-4.61, P = .024), preoperative transfusions (OR 2.91, 95% CI: 1.02-8.29, P = .049), and preoperative infection/sepsis (OR 2.67, 95% CI: 1.18-6.08, P = .02). Our model demonstrates very strong predictive capabilities, with an area under the receiver operating characteristic curve of 0.7447. Steroid use is associated with a significant increased risk of 30-d mortality in surgical metastatic spine tumor patients with disseminated disease. These findings warrant further investigation in controlled experimental environments.


Rural children active trachoma risk factors and their interactions.
Trachoma is a serious public health problem in rural Ethiopia. The aim of this investigation was to provide in-depth statistical analysis of the risk factors associated with active trachoma among children of age 1-9 years of Kedida Gamela district, in Ethiopia. A community based cross-sectional survey of trachoma was conducted in six selected rural kebeles of Kedida Gamela district, in Ethiopia from June 10-25, 2014. A total of 377 children (ages 1-9 years) were included in the study using two stage cluster sampling. All children were examined for trachoma by nurse data collectors supervised by ophthalmic supervisors using the WHO simplified clinical grading system. Ordinal survey logistic regression model was used to identify risk factors. Data analysis was done using SAS version 9.3. The best fit proportional odds model was identified to be the main effects and two-way and three-way interactios. Keeping cattle in the house was found to have a protective effect (OR=0.138, p-value=0.0003). The household wealth will have a more protective effect if the child attends school. Washing face with soap and water once a day has equivalent protective effect as washing face three-or-more times a day with water only. The 2-way and 3-way significant interactions effects unfolded some of the controversies derived from similar studies on trachoma risk factors. The findings would suggest integrated effort to address two or three factors simultaneously is more fruitful than any novel intervention targeted to address a single risk factor.


Rodents as sentinels for the prevalence of tick-borne encephalitis virus.
Tick-borne encephalitis virus (TBEV) causes one of the most important flavivirus infections of the central nervous system, affecting humans in Europe and Asia. It is mainly transmitted by the bite of an infected tick and circulates among them and their vertebrate hosts. Until now, TBE risk analysis in Germany has been based on the incidence of human cases. Because of an increasing vaccination rate, this approach might be misleading, especially in regions of low virus circulation. To test the suitability of rodents as a surrogate marker for virus spread, laboratory-bred Microtus arvalis voles were experimentally infected with TBEV and analyzed over a period of 100 days by real-time (RT)-quantitative polymerase chain reaction. Further, the prevalence of TBEV in rodents trapped in Brandenburg, a rural federal state in northeastern Germany with autochthonous TBE cases, was determined and compared with that in rodents from German TBE risk areas as well as TBE nonrisk areas. In experimentally infected M. arvalis voles, TBEV was detectable in different organs for at least 3 months and in blood for 1 month. Ten percent of all rodents investigated were positive for TBEV. However, in TBE risk areas, the infection rate was higher compared with that of areas with only single human cases or of nonrisk areas. TBEV was detected in six rodent species: Apodemus agrarius, Apodemus flavicollis, Apodemus sylvaticus, Microtus agrestis, Microtus arvalis, and Myodes glareolus. M. glareolus showed a high infection rate in all areas investigated. The infection experiments proved that TBEV can be reliably detected in infected M. arvalis voles. These voles developed a persistent TBE infection without clinical symptoms. Further, the study showed that rodents, especially M. glareolus, are promising sentinels particularly in areas of low TBEV circulation.


Plasma membrane targeting of chimeric intracisternal A-type particle polyproteins leads to particle release and specific activation of the viral proteinase.
Retrovirus morphogenesis involves assembly of structural Gag polyproteins with subsequent budding from the plasma membrane, followed by proteolytic cleavage by the viral proteinase (PR) and extracellular maturation to the infectious virion. Intracisternal A-type particles (IAPs) are defective retroviruses that assemble and bud at the membranes of the endoplasmic reticulum (ER), where they remain as immature particles consisting exclusively of uncleaved polyproteins. To analyze requirements for intracellular polyprotein transport and PR activation, we constructed deletion and substitution mutations in the IAP gag gene, including the putative ER-targeting signal. Mutant polyproteins were transported to various intracellular locations, including the nucleus, the cytoplasm, the ER, and the plasma membrane. Interestingly, assembly of capsid-like particle structures occurred at almost all sites. However, only those polyproteins transported to the plasma membrane were efficiently and specifically cleaved by viral PR, with cleavage occurring predominantly within the virus particle. Thus, at least in the experimental system presented here, retroviral particle assembly can occur at almost any location within the cell, while polyprotein processing and, consequently, virion maturation are confined to a specific cellular site. These results suggest that a factor restricted to the plasma membrane is required to trigger PR activation and maturation of infectious retroviruses.


Combinational effect of low-dose oral corticosteroid and mizoribine for ulcerative colitis in a patient with systemic sclerosis.
The combinational effect of oral corticosteroid and mizoribine for ulcerative colitis is presented in a patient with systemic sclerosis (SSc). A 64-year-old woman came to our clinic complaining of a 30-year history of Raynaud's phenomenon. She had past history of ulcerative colitis with the continued medication of mesalazine without success. She was presented with sclerodactyly and finger joint swelling. She also showed epigastric discomfort. Laboratory study showed positive anti-nuclear antibody and positive anti-centromere antibody. Histological examination showed mild perivascular mononuclear cell infiltrates in the whole dermis and increased deposition of collagen fibers in the middle and lower dermis. Chest X-ray film showed mild bibasilar pulmonary fibrosis. An upper gastrointestinal series study showed reflux esophagitis and atrophic gastritis. These findings led to the diagnosis of systemic sclerosis (limited type) complicated with ulcerative colitis. Treatment with oral corticosteroid (5 mg/day) and mizoribine (150 mg/day) in the morning was started. She showed remarkable improvement for sclerodactyly and lower intestinal bleeding stopped after 6 months. She is under the same treatment without exaggeration and adverse effect of the drug until now.


Electrolysis on a Chip with Tunable Thin Film Nanostructured PGM Electrocatalysts Generated from Self-Assembled Block Copolymer Templates.
Self-assembled block copolymers are promising templates for fabricating thin film materials with tuned periodic feature sizes and geometry at the nanoscale. Here, a series of nanostructured platinum and iridium oxide electrocatalysts templated from poly(styrene)-block-poly(vinyl pyridine) (PSbPVP) block copolymers via an incipient wetness impregnation (IWI) pathway is reported. Both nanowire and nanocylinder electrocatalysts of varying feature sizes are assessed and higher catalyst loadings are achieved by the alkylation of the pyridine moieties in the PVP block prior to IWI. Electrocatalyst evaluations featuring hydrogen pump and water electrolysis demonstrations are carried out on interdigitated electrode (IDE) chips flexible with liquid supporting electrolytes and thin film polymer electrolytes. Notably, the mass activities of the nanostructured electrocatalysts from alkylated block copolymer templates are 35%-94% higher than electrocatalysts from non-alkylated block copolymer templates. Standing cylinder nanostructures lead to higher mass activities than lamellar variants despite their not having the largest surface area per unit catalyst loading demonstrating that mesostructure architectures have a profound impact on reactivity. Overall, IDE chips with model thin film electrocatalysts prepared from self-assembled block copolymers offer a high-throughput experimental method for correlating electrocatalyst nanostructure and composition to electrochemical reactivity.


Interferon-inducible LY6E Protein Promotes HIV-1 Infection.
LY6E is a glycosylphosphatidylinositol-anchored, IFN-inducible protein that regulates T lymphocytes proliferation, differentiation, and development. Single-nucleotide polymorphism rs2572886 in the LY6 family protein locus has been shown to associate with accelerated progression to AIDS. In this study, we show that LY6E promotes HIV, type 1 (HIV-1) infection by enhancing viral entry and gene expression. Knockdown of LY6E in human peripheral blood mononuclear, SupT1, and THP-1 cells diminishes HIV-1 replication. Virion-cell and cell-cell fusion experiments revealed that LY6E promotes membrane fusion of the viral entry step. Interestingly, we find that LTR-driven HIV-1 gene expression is also enhanced by LY6E, suggesting additional roles of LY6E in HIV-1 replication. HIV-1 infection induces LY6E expression in human peripheral blood mononuclear cells, concomitant with increased production of type I IFN and some classical IFN-stimulated genes. Altogether, our results demonstrate that IFN-inducible LY6E promotes HIV-1 entry and replication and highlight a positive regulatory role of IFN-induced proteins in HIV-1 infection. Our work emphasizes the complexity of IFN-mediated signaling in HIV-host interaction and AIDS pathogenesis.


Mechanisms of apigenin-7-glucoside as a hepatoprotective agent.
Ixeris chinesis (Thunb.) Ankai has been used as a Chinese folk medicine, but only scanty information is available on the physiological and biochemical functions of the compounds extracted from I. chinesis. In the present study the effects of apigenin-7-glucoside (APIG) isolated from I. chinesis against liver injury caused by carbon tetrachloride (CCl4) were investigated. The contents of malondialdehyde (MDA), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), and reduced glutathione (GSH) were evaluated by spectrophotography. The content of 8-Hydroxydeoxyguanosine (8-OHdG) was measured with high-performance liquid chromatography (HPLC) equipped with electrochemical and UV detection methods. The antioxidant activity of APIG was evaluated using chemiluminescence single photon counting technology. CCl4 significantly increased the enzyme activities of GPT and GOT in blood serum, as well as the level of MDA and 8-OHdG in liver tissue, and decreased the levels of GSH. Pretreatment with APIG was able not only to suppress the elevation of GPT, GOT, MDA and 8-OHdG, and inhibit the reduction of GSH in a dose-dependent manner in vivo, but also to reduce the damage of hepatocytes in vitro. On the other hand, we also found that APIG had strong antioxidant activity against reactive oxygen species (ROS) in vitro in a concentration-dependent manner. The hepatoprotective activity of APIG is possibly due to its antioxidant properties, acting as scavengers of ROS. These results obtained in vivo and in vitro suggest that APIG has protective effects against hepatic oxidative injury induced by chemicals. Further studies on the pharmaceutical functions and immunological responses of APIG may help its clinical application.


Construction of a cloning system for the mass production of a virus-binding protein specific for poliovirus type 1.
In our previous study, virus-binding proteins (VBPs) demonstrating the ability to strongly bind poliovirus type 1 (PV1) were recovered from a bacterial culture derived from activated sludge. The isolated VBPs would be useful as viral adsorbents for water and wastewater treatments. The VBP gene of activated sludge bacteria was isolated, and the cloning system of the VBP was established. The isolation of the VBP gene from DNA libraries for activated sludge bacteria was achieved with the colony hybridization technique. The sequence of the VBP gene consisted of 807 nucleotides encoding 268 amino acids. Fifteen amino acid sequences were retrieved from 2,137,877 sequences by a homology search using the BLAST server at the National Center for Biotechnology Information. The protein encoded in the isolated genome was considered to be a newly discovered protein from activated sludge culture, because any sequences in protein databases were not perfectly matched with the sequence of the VBP. It was confirmed that Escherichia coli BL21 transformed by pRSET carrying the isolated VBP gene could extensively produce the VBP clones. Enzyme-linked immunosorbent assay (ELISA) revealed that the VBP clone exhibited the binding ability with intact particles of PV1. The equilibrium binding constant between PV1 and VBP in the ELISA well was estimated to be 2.1 x 10(7) (M(-1)), which also indicated that the VBP clones have a high affinity with the PV1 particle. The VBP cloning system developed in this study would make it possible to produce a mass volume of VBPs and to utilize them as a new material of the specific adsorbent in several technologies, including virus removal, concentration, and detection.


Development and validation of a protocol for optimizing the use of paraffin blocks in molecular epidemiological studies: The example from the HPV-AHEAD study.
Worldwide use of formalin-fixed paraffin-embedded blocks (FFPE) is extensive in diagnosis and research. Yet, there is a lack of optimized/standardized protocols to process the blocks and verify the quality and presence of the targeted tissue. In the context of an international study on head and neck cancer (HNC)-HPV-AHEAD, a standardized protocol for optimizing the use of FFPEs in molecular epidemiology was developed and validated. First, a protocol for sectioning the FFPE was developed to prevent cross-contamination and distributed between participating centers. Before processing blocks, all sectioning centers underwent a quality control to guarantee a satisfactory training process. The first and last sections of the FFPEs were used for histopathological assessment. A consensus histopathology evaluation form was developed by an international panel of pathologists and evaluated for four indicators in a pilot analysis in order to validate it: 1) presence/type of tumor tissue, 2) identification of other tissue components that could affect the molecular diagnosis and 3) quality of the tissue. No HPV DNA was found in sections from empty FFPE generated in any histology laboratories of HPV-AHEAD consortium and all centers passed quality assurance for processing after quality control. The pilot analysis to validate the histopathology form included 355 HNC cases. The form was filled by six pathologists and each case was randomly assigned to two of them. Most samples (86%) were considered satisfactory. Presence of >50% of invasive carcinoma was observed in all sections of 66% of cases. Substantial necrosis (>50%) was present in <2% of samples. The concordance for the indicators targeted to validate the histopathology form was very high (kappa > 0.85) between first and last sections and fair to high between pathologists (kappa/pabak 0.21-0.72). The protocol allowed to correctly process without signs of contamination all FFPE of the study. The histopathology evaluation of the cases assured the presence of the targeted tissue, identified the presence of other tissues that could disturb the molecular diagnosis and allowed the assessment of tissue quality.


Pneumococcal pneumonia and endotoxemia: An experimental and clinical reappraisal.
Circulating endotoxins could result from bacterial digestive translocation during sepsis, thus contributing to uncontrolled systemic inflammation, leading in turn to organ dysfunction. We addressed this issue in the setting of severe pneumococcal pneumonia. Endotoxemia was measured in a clinically relevant rabbit model of ventilated pneumococcal pneumonia and in 110 patients with bacteraemic pneumonia, using a patented mass spectrometry (LC-MS/MS) method for detection of 3-OH fatty acids (C10, C12, C14, C16 and C18), which are molecules bound to the lipid A motif of LPS. Whereas higher levels of systemic inflammation and organ dysfunctions were found, there was no significant difference in lipopolysaccharide concentrations when infected rabbits were compared to non-infected ones, or when patients were compared to healthy volunteers. Seemingly, endotoxins do not drive the overwhelming inflammation associated with severe forms of pneumococcal pneumonia.


Fluctuations around mean walking behaviors in diluted pedestrian flows.
Understanding and modeling the dynamics of pedestrian crowds can help with designing and increasing the safety of civil facilities. A key feature of a crowd is its intrinsic stochasticity, appearing even under very diluted conditions, due to the variability in individual behaviors. Individual stochasticity becomes even more important under densely crowded conditions, since it can be nonlinearly magnified and may lead to potentially dangerous collective behaviors. To understand quantitatively crowd stochasticity, we study the real-life dynamics of a large ensemble of pedestrians walking undisturbed, and we perform a statistical analysis of the fully resolved pedestrian trajectories obtained by a yearlong high-resolution measurement campaign. Our measurements have been carried out in a corridor of the Eindhoven University of Technology via a combination of Microsoft Kinect 3D range sensor and automatic head-tracking algorithms. The temporal homogeneity of our large database of trajectories allows us to robustly define and separate average walking behaviors from fluctuations parallel and orthogonal with respect to the average walking path. Fluctuations include rare events when individuals suddenly change their minds and invert their walking directions. Such tendency to invert direction has been poorly studied so far, even if it may have important implications on the functioning and safety of facilities. We propose a model for the dynamics of undisturbed pedestrians, based on stochastic differential equations, that provides a good agreement with our field observations, including the occurrence of rare events.


Benchmark-Based Reference Model for Evaluating Botnet Detection Tools Driven by Traffic-Flow Analytics.
Botnets are some of the most recurrent cyber-threats, which take advantage of the wide heterogeneity of endpoint devices at the Edge of the emerging communication environments for enabling the malicious enforcement of fraud and other adversarial tactics, including malware, data leaks or denial of service. There have been significant research advances in the development of accurate botnet detection methods underpinned on supervised analysis but assessing the accuracy and performance of such detection methods requires a clear evaluation model in the pursuit of enforcing proper defensive strategies. In order to contribute to the mitigation of botnets, this paper introduces a novel evaluation scheme grounded on supervised machine learning algorithms that enable the detection and discrimination of different botnets families on real operational environments. The proposal relies on observing, understanding and inferring the behavior of each botnet family based on network indicators measured at flow-level. The assumed evaluation methodology contemplates six phases that allow building a detection model against botnet-related malware distributed through the network, for which five supervised classifiers were instantiated were instantiated for further comparisons-Decision Tree, Random Forest, Naive Bayes Gaussian, Support Vector Machine and K-Neighbors. The experimental validation was performed on two public datasets of real botnet traffic-CIC-AWS-2018 and ISOT HTTP Botnet. Bearing the heterogeneity of the datasets, optimizing the analysis with the Grid Search algorithm led to improve the classification results of the instantiated algorithms. An exhaustive evaluation was carried out demonstrating the adequateness of our proposal which prompted that Random Forest and Decision Tree models are the most suitable for detecting different botnet specimens among the chosen algorithms. They exhibited higher precision rates whilst analyzing a large number of samples with less processing time. The variety of testing scenarios were deeply assessed and reported to set baseline results for future benchmark analysis targeted on flow-based behavioral patterns.


Identification of an adult stem/progenitor cell-like population in the human thyroid.
There is evidence that tissue-specific stem cells reside in certain adult tissues. Their specific properties remain elusive, because they are rare and heterogeneous in parent tissues; furthermore, technical difficulties have been encountered in the identification and characterization of their progeny. The aim of this study was to isolate stem/progenitor cells from the human thyroid. We devised a method based on the enzymatic digestion of fresh surgical thyroid specimens, followed by culture of cells in the presence of epidermal growth factor and basic fibroblast growth factor. We also used markers that identify and characterize these cells. Spheroids with self-replicative potential were obtained from all thyroid specimens. The isolated population contained a subset of CD34+ CD45- cells and it was able, in differentiation conditions, to generate follicles with thyroid hormonal production. In support of the plasticity concept, we obtained evidence that, when most freshly isolated spheroids were co-cultured with a neuroblastoma cell line, they produced progeny expressing the neuronal marker beta-tubulin III. Spheroids were also able to undergo adipogenic differentiation when cultured in adipogenic medium. We conclude that a predominant functional type of stem/progenitor cell exists within the thyroid, with an intrinsic ability to generate thyroidal cells and the potential to produce non-thyroidal cells.


A randomised controlled trial of written information: the effect on parental non-concordance with occlusion therapy.
It is well documented that non-concordance with occlusion therapy is both substantial and a major factor leading to treatment failure. Parental understanding in previous work has been found to be poor in key areas such as the critical period and effect of age on prognosis. Research in other areas of medicine has shown that the level of understanding can have a direct effect on the level of concordance. The aims of this study were to assess the ability of educational material in the form of a leaflet, to improve parental understanding of amblyopia and occlusion, and subsequently increase concordance. Parents of children aged between 1 and 7 years receiving a minimum of 1 hour of occlusion for amblyopia were recruited. A randomised controlled trial was undertaken where, on inclusion, the patients were randomised into a leaflet group, whose parents were issued with written educational material, and a control group whose parents did not receive the written information. Patients were paired and matched for age (<2 years difference) and amount of prescribed occlusion (no more than 1 hour difference). Concordance was monitored by a parental diary and knowledge and parental reasons for non-concordance were assessed by a questionnaire. Concordance was analysed by means of a concordance index and by calculating the proportion of non-concordant parents by setting a threshold of concordance at 80%. Parental knowledge was significantly greater in the leaflet group (88% had complete knowledge) compared to the control group (49% had complete knowledge) (p <0.001). There were also differences between the groups in the area of the treatment regimen, with errors only occurring in the control group (three patients occluded the incorrect eye), but this did not reach statistical significance. Concordance was significantly greater in the leaflet group (mean concordance index 0.85) compared to the non-leaflet group (mean concordance index (0.71) (p <0.001). Comparison of the proportion of non-concordant parents was also statistically different (p <0.005) at 0.23 (95% CI 0.13 to 0.35) for the leaflet group compared to 0.54 (95% CI 0.41 to 0.67) for the control group. A large proportion of patients would benefit by increasing parental knowledge in key areas such as the critical period, importance of occlusion, and potential negative consequences of not treating amblyopia. Written information is a simple, inexpensive, easy to implement, yet effective method of improving parental understanding and subsequent concordance.


Evolving guidelines in the use of topical nonsteroidal anti-inflammatory drugs in the treatment of osteoarthritis.
Nonsteroidal anti-inflammatory drugs (NSAIDs) are a standard treatment for osteoarthritis (OA), but the use of oral NSAIDs has been linked to an elevated risk for cardiovascular and gastrointestinal adverse events and renal toxicity. Topical NSAIDs are thought to afford efficacy that is comparable to oral formulations while reducing widespread systemic drug exposure, which may provide a benefit in terms of safety and tolerability. As a result, European treatment guidelines have, for many years, recommended the use of topical NSAIDs as a safe and effective treatment option for OA. Following the recent approval of several topical NSAID formulations by the US Food and Drug Administration, US treatment guidelines are increasingly recommending the use of topical NSAIDs as an alternative therapy and, in some cases, as a first-line option for OA. This commentary summarizes OA treatment guidelines that are currently available and discusses their potential evolution with regard to the increased inclusion of topical NSAIDs.


Neurogenic bladder in Rocky Mountain spotted fever.
A previously healthy 29-year-old man who had verified Rocky Mountain spotted fever had a flaccid neurogenic bladder that resolved as he recovered from his infection. The neurologic complications of this disease are presumed to be related to vasculitis with resulting ischemia or infarction of nervous tissue.


Actively personalized vaccination trial for newly diagnosed glioblastoma.
Patients with glioblastoma currently do not sufficiently benefit from recent breakthroughs in cancer treatment that use checkpoint inhibitors1,2. For treatments using checkpoint inhibitors to be successful, a high mutational load and responses to neoepitopes are thought to be essential3. There is limited intratumoural infiltration of immune cells4 in glioblastoma and these tumours contain only 30-50 non-synonymous mutations5. Exploitation of the full repertoire of tumour antigens-that is, both unmutated antigens and neoepitopes-may offer more effective immunotherapies, especially for tumours with a low mutational load. Here, in the phase I trial GAPVAC-101 of the Glioma Actively Personalized Vaccine Consortium (GAPVAC), we integrated highly individualized vaccinations with both types of tumour antigens into standard care to optimally exploit the limited target space for patients with newly diagnosed glioblastoma. Fifteen patients with glioblastomas positive for human leukocyte antigen (HLA)-A*02:01 or HLA-A*24:02 were treated with a vaccine (APVAC1) derived from a premanufactured library of unmutated antigens followed by treatment with APVAC2, which preferentially targeted neoepitopes. Personalization was based on mutations and analyses of the transcriptomes and immunopeptidomes of the individual tumours. The GAPVAC approach was feasible and vaccines that had poly-ICLC (polyriboinosinic-polyribocytidylic acid-poly-L-lysine carboxymethylcellulose) and granulocyte-macrophage colony-stimulating factor as adjuvants displayed favourable safety and strong immunogenicity. Unmutated APVAC1 antigens elicited sustained responses of central memory CD8+ T cells. APVAC2 induced predominantly CD4+ T cell responses of T helper 1 type against predicted neoepitopes.


Effects of thymoquinone in prevention of experimental contrast-induced nephropathy in rats.
This study aimed to show the effects of thymoquinone, which is known for its antioxidant, anti-inflammatory, and renal protective effects in contrast-induced nephropathy. This is an experimental study in rats. 7 groups were included within the scope of our study: sham-vehicle (n=3), premedication-control (n=6), model (n=6), isolated thymoquinone (n=3+3), low-dose thymoquinone (n=6), and high-dose thymoquinone (n=7). In addition to 48 hr of water deprivation, we pre-medicated the rats with intra-peritoneal indomethacin and L-NAME administration. After premedication, 12.5 ml/kg dose of a high osmolar contrast agent-diatrizoat (Urografin %76) was administrated. Thymoquinone was administrated in two different doses of 1 mg/kg and 1.75 mg/kg for four days intraperitoneally. Renal functions, histopathological differences, oxidative stress parameters, and inflammatory indicators of rats were evaluated at the end of the study. Significant decreases were observed in levels of serum creatinine and serum BUN with low-dose thymoquinone (1 mg/kg) administration. In light microscopy, significantly less histopathological damage was observed in the low-dose thymoquinone group compared to the contrast agent group. While high-dose thymoquinone is accepted as ineffective biochemically, toxic evidence was identified histopathologically. There were no significant differences between M and TA groups for serum MDA and SOD levels, which were compared to evaluate oxidative stress (P:0.99, P:0.98; respectively). TNF-α, iNOS, and NF-кB gene expressions were not significantly different between all groups (P:0.748, P:0.531, P:0.910; respectively). This experimental study has demonstrated for the first time the protective effect of the TQ substance for CIN in 1 mg/kg dose, in the accompaniment of biochemical and histopathological data in rats.


Cation effects on rotational dynamics of anions and water molecules in alkali (Li+, Na+, K+, Cs+) thiocyanate (SCN-) aqueous solutions.
Waiting time dependent rotational anisotropies of SCN(-) anions and water molecules in alkali thiocyanate (XSCN, X = Li, Na, K, Cs) aqueous solutions at various concentrations were measured with ultrafast infrared spectroscopy. It was found that cations can significantly affect the reorientational motions of both water molecules and SCN(-) anions. The dynamics are slower in a solution with a smaller cation. The reorientational time constants follow the order of Li(+) > Na(+) > K(+) ~/= Cs(+). The changes of rotational time constants of SCN(-) at various concentrations scale almost linearly with the changes of solution viscosity, but those of water molecules do not. In addition, the concentration-dependent amplitudes of dynamical changes are much more significant in the Li(+) and Na(+) solutions than those in the K(+) and Cs(+) solutions. Further investigations on the systems with the ultrafast vibrational energy exchange method and molecular dynamics simulations provide an explanation for the observations: the observed rotational dynamics are the balanced results of ion clustering and cation/anion/water direct interactions. In all the solutions at high concentrations (>5 M), substantial amounts of ions form clusters. The structural inhomogeneity in the solutions leads to distinct rotational dynamics of water and anions. The strong interactions of Li(+) and Na(+) because of their relatively large charge densities with water molecules and SCN(-) anions, in addition to the likely geometric confinements because of ion clustering, substantially slow down the rotations of SCN(-) anions and water molecules inside the ion clusters. The interactions of K(+) and Cs(+) with water or SCN(-) are much weaker. The rotations of water molecules inside ion clusters of K(+) and Cs(+) solutions are not significantly different from those of other water species so that the experimentally observed rotational relaxation dynamics are only slightly affected by the ion concentrations.


[Determination of neurometabolism in patients with mild and moderate cognitive impairments].
To estimate the diagnostic value of neuropower mapping (NPM) and positron emission tomography (PET) and their relationships in the evaluation of neurometabolism in patients with predementia cognitive impairments. MATERIAL AND METHODS. The investigation enrolled 125 patients wvith clinical signs of cognitive impairments. The patients were divided into 4 groups: 1) predementia cognitive impairments (PCI) in the presence of cerebrovascular disease (CVD); 2) PCI in the presence of psychoautonomic syndrome (PAS); 3) PCI in the presence of sequels to acute cerebrovascular accident (SCVA); 4) PCI in the presence of toxic, dysmetabolic, hypoxic encephalopathies (TDHEP). All the patients underwent NPM and 55 patients had PET. NPM and PET revealed neurometabolic disturbances in 100 and 46.3% of cases, respectively. The sensitivity of NPM was significantly higher than that of PET. The highest number of diffuse and diffuse-focal hypometabolism cases (n = 19 (63.3%)) were seen in the CVD and SCVA groups. CONCLUSION. 1. NPM and PET assess two aspects of brain neurometabolism. 2. PET hypometabolism characterizes either structural organic abnormalities in the brain in the presence SCVA or initial manifestations of the neurodegenerative process. 3. NPM readings characterize the activity, strength of metabolic responses, that is to say, cerebral reactivity. 4. PET readings are important in determining the prognosis of disease and NPM ones are essential in defining drug treatment policy.


Emerging options with coxib therapy.
Future clinical applications of cyclooxygenase (COX)-2-selective inhibitors (coxibs) are likely to extend beyond their current use as oral analgesics in high-risk arthritis patients. The clinical utility of coxibs for the treatment of Alzheimer's disease (AD) is under investigation. Epidemiological surveys, preclinical studies, and preliminary clinical trials with nonsteroidal anti-inflammatory drugs (NSAIDs) have suggested that inflammatory mechanisms play a role in the neurodegeneration of AD. Clinical trials are currently being conducted to determine the effect of coxibs on the rate of AD progression. The use of coxibs as chemopreventive agents in colorectal cancer (CRC) is also under investigation. The chemopreventive benefits of coxibs to promote cell death (apoptosis) and inhibit angiogenesis in CRC have been shown in tumor cell lines and in animal and human models. In addition, palliative care clinicians and oncologists are increasingly including coxibs in their management of cancer pain. Coxibs are utilized for their opioid-sparing effect in the management of cancer pain, without impairing wound healing, or promoting bleeding diathesis (antiplatelet effects) or adverse gastrointestinal effects in patients receiving chemotherapy or radiation treatment.


Interaction of pathogenic yeasts with phagocytes: survival, persistence and escape.
Pathogenic yeasts, either from the environment or the normal flora, have to face phagocytic cells that constitute the first line of defence during infection. In order to evade or counteract attack by phagocytes, pathogenic yeasts have acquired a repertoire of strategies to survive, colonize and infect the host. In this review we focus on the interaction of yeasts, such as Candida, Histoplasma or Cryptococcus species, with macrophages or neutrophils. We discuss strategies used by these fungi to prevent phagocytosis or to counteract phagocytic activities. We go on to describe the strategies that permit intracellular survival within phagocytes and that may eventually lead to damage of and escape from the phagocyte.


Multilocus sequence typing of Streptococcus uberis provides sensitive and epidemiologically relevant subtype information and reveals positive selection in the virulence gene pauA.
Control of the bovine mastitis pathogen Streptococcus uberis requires sensitive and epidemiologically meaningful subtyping methods that can provide insight into this pathogen's epidemiology and evolution. Development of a multilocus sequence typing (MLST) scheme based on six housekeeping and virulence genes allowed differentiation of 40 sequence types among 50 S. uberis isolates from the United States (n = 30) and The Netherlands (n = 20). MLST was more discriminatory than EcoRI or PvuII ribotyping and provided subtype data with better epidemiological relevance, e.g., by discriminating isolates with identical ribotypes obtained from different farms. Phylogenetic analyses of MLST data revealed indications of reticulate evolution between genes, preventing construction of a core phylogeny based on concatenated DNA sequences. However, all individual gene phylogenies clearly identified a distinct pauA-negative subtaxon of S. uberis for which housekeeping alleles closely resembled those of Streptococcus parauberis. While the average GC content for five genes characterized was between 0.38 and 0.40, pauA showed a considerably lower GC content (0.34), suggesting acquisition through horizontal transfer. pauA also showed a higher nonsynonymous/synonymous rate ratio (dN/dS) (1.2) compared to the other genes sequenced (dN/dS < 0.12), indicating positive selection in this virulence gene. In conclusion, our data show that (i) MLST provides for highly discriminatory and epidemiologically relevant subtyping of S. uberis; (ii) S. uberis has a recombinatorial population structure; (iii) phylogenetic analysis of MLST data reveals an S. uberis subtaxon resembling S. parauberis; and (iv) horizontal gene transfer and positive selection contribute to evolution of certain S. uberis genes, such as the virulence gene pauA.


Activation of Arp2/3 complex-mediated actin polymerization by cortactin.
Cortactin, a filamentous actin (F-actin)-associated protein and prominent substrate of Src, is implicated in progression of breast tumours through gene amplification at chromosome 11q13. However, the function of cortactin remains obscure. Here we show that cortactin co-localizes with the Arp2/3 complex, a de novo actin nucleator, at dynamic particulate structures enriched with actin filaments. Cortactin binds directly to the Arp2/3 complex and activates it to promote nucleation of actin filaments. The interaction of cortactin with the Arp2/3 complex occurs at an amino-terminal domain that is rich in acidic amino acids. Mutations in a conserved amino-acid sequence of DDW abolish both the interaction with the Arp2/3 complex and complex activation. The N-terminal domain is not only essential but also sufficient to target cortactin to actin-enriched patches within cells. Interestingly, the ability of cortactin to activate the Arp2/3 complex depends on an activity for F-actin binding, which is almost 20-fold higher than that of the Arp2/3 complex. Our data indicate a new mechanism for activation of actin polymerization involving an enhanced interaction between the Arp2/3 complex and actin filaments.


Radiologic differential diagnosis. Radiologic pattern: solitary cavity.
The differential diagnosis of a left lower lobe cavity in this young patient with a history of productive cough should include hiatal hernia, pulmonary abscess, bronchiectatic cyst and bronchopulmonary sequestration. Hiatal hernia should be ruled out by barium swallow; acute pulmonary abscess by the lack of a history suggestive of a necrotizing pneumonia; bronchiectasis by bronchogram; and intralobar bronchopulmonary sequestration should be confirmed by aortography.


Conoscopy of chiral smectic liquid crystal cells.
The conoscopic method for investigating the optical properties of a liquid crystal cell is studied with the aim of determining the effects of the approximations used in the calculation on the results. We confirm that the chiral liquid crystal cell forming a helical structure can be regarded as a single biaxial plate for analyzing the conoscopic image only if the helical pitch is less than several multiples of the wavelength of light. This approximation implies that the square of the refractive index along a direction is averaged over all the layers. An incorrectly chosen value for one of the principal refractive indices to be used in the analysis of the conoscopic data can lead to an incorrect conclusion, especially for the case when the wavelength dispersion of the refractive index is neglected. A thicker cell and a longer wavelength of the incident light can minimize these limitations of the conoscopic method. We propose a novel simulation method to find the molecular distribution in a liquid crystal cell based on the average-refractive-index approximation and the conoscopic data. This is shown to be a fast, more efficient, and useful method for estimating the director distributions.


Comparison of two point-of-care international normalized ratio devices and laboratory method.
: A number of factors contribute to interlaboratory variation of international normalized ratio (INR) results. This variability can lead to differences in clinical decision-making. This prospective cohort study evaluated the accuracy, precision, and clinical decision-making variability of two point-of-care (POC) devices (CoaguChek XS and the Coag-Sense) to a clinical laboratory method of INR measurement as the 'reference' and by using the laboratory's accuracy standard (±25% of the reference INR). Study subjects included adults taking warfarin undergoing a POC INR with each device and a laboratory INR on the same day and evaluated differences in clinical decision-making using a warfarin nomogram. A correction factor was derived for each POC method to estimate the corresponding laboratory INR for POC INRs more than 3.0. A total of 100 patient encounters (76 unique patients) included INR testing by all three methods. Both POC devices demonstrated excellent precision across the entire range (r = 0.98), but poor accuracy relative to the laboratory standard for POC INRs greater than 3.0 (accuracy CoaguChek 19% and Coag-Sense 35%). A correction factor applied to POC INRs greater than 3.0 improved accuracy to 99% for both devices. Applying the correction factor also significantly reduced differences in clinical decision-making (49-28% for CoaguChek and 56-40% for Coag-Sense, P < 0.001). Both POC devices demonstrated poor accuracy for INRs over 3.0. A device-specific, institution-specific correction factor applied to POC INRs greater than 3.0 can provide better accuracy of INR measurement and can significantly reduce variability in warfarin dosing decisions. More study is needed to assess impact on clinical adverse events.


Satisfactory Analgesia with Minimal Emesis in Day Surgeries (SAME DayS): a protocol for a randomised controlled trial of morphine versus hydromorphone.
There has been an exponential increase in the number of ambulatory surgeries (AS). Pain and postoperative nausea vomiting (PONV) affects the recovery, discharge and overall satisfaction of patients having AS. Opioids remain the primary modality for moderate to severe pain. Since there is no perfect opioid, physicians should ideally use the opioid that optimally balances benefits and risks. Present decisions on the choice between morphine (M) and hydromorphone (HM) are based on individual experience and observation. Our primary objective is to compare the proportion of patients having AS achieving satisfactory analgesia without significant PONV when using M compared with HM. Secondarily we will compare the proportion of patients with adverse events, analgesic used, patient satisfaction, time to discharge and postdischarge symptoms. This is a two-arm, multicentre, parallel group, randomised controlled trial of 400 patients having AS. Eligible patients undergoing AS of the abdominal and pelvic regions with a potential to cause moderate to severe pain will be recruited in the preoperative clinic. Using a computer-generated randomization, with a 1:1 allocation ratio, patients will be randomised to M or HM. Patients, healthcare providers and research personnel will be blinded. Study interventions will be administered in the recovery using equianalgesic doses of M or HM in concealed syringes. Patients will be followed in hospital and up to 3 months. Intention-to-treat approach will be used for analysis. This study has been approved by the Hamilton integrated research ethics board. We plan to publish our trial findings and present our findings at scientific meetings. NCT02223377; Pre-results.


[Current treatment options for recurrent/metastatic renal cell carcinoma (mRCC)].
Renal cell carcinoma is a common malignant tumour, whose incidence is steadily increasing. Whilst operative techniques have recently improved, including possibilities of renal preservation and minimally invasive surgery, no use has been shown for adjuvant treatment strategies. When local therapies are not suitable in relapsed or metastatic disease, medical drug treatment is indicated. The first immunologic treatment for renal cell carcinoma consisted of cytokines and overall survival amounted to a median of 13 months. Over the last decade, tyrosine kinase inhibitors (TKI), which inhibit the tumour vasculature, dominated the therapeutic area and raised the clinical discussion about the best sequential pattern. More recently, checkpoint inhibitors (CPI) have set a new standard as therapeutic tools in renal cell cancer. Modern first-line therapies consist of checkpoint inhibitor-based combinations, which have shifted the median overall survival expectation to >40 months. However, tyrosine kinase inhibitors have retained their therapeutic value as combination partners and also as monotherapy in individual cases. One combined immunotherapy and 3 combinations of a checkpoint inhibitor and a TKI are currently approved in Germany.


Integrated array-comparative genomic hybridization and expression array profiles identify clinically relevant molecular subtypes of glioblastoma.
Glioblastoma, the most aggressive primary brain tumor in humans, exhibits a large degree of molecular heterogeneity. Understanding the molecular pathology of a tumor and its linkage to behavior is an important foundation for developing and evaluating approaches to clinical management. Here we integrate array-comparative genomic hybridization and array-based gene expression profiles to identify relationships between DNA copy number aberrations, gene expression alterations, and survival in 34 patients with glioblastoma. Unsupervised clustering on either profile resulted in similar groups of patients, and groups defined by either method were associated with survival. The high concordance between these separate molecular classifications suggested a strong association between alterations on the DNA and RNA levels. We therefore investigated relationships between DNA copy number and gene expression changes. Loss of chromosome 10, a predominant genetic change, was associated not only with changes in the expression of genes located on chromosome 10 but also with genome-wide differences in gene expression. We found that CHI3L1/YKL-40 was significantly associated with both chromosome 10 copy number loss and poorer survival. Immortalized human astrocytes stably transfected with CHI3L1/YKL-40 exhibited changes in gene expression similar to patterns observed in human tumors and conferred radioresistance and increased invasion in vitro. Taken together, the results indicate that integrating DNA and mRNA-based tumor profiles offers the potential for a clinically relevant classification more robust than either method alone and provides a basis for identifying genes important in glioma pathogenesis.


Relative effects of increment and pedestal duration on the detection of intensity increments.
The detection of a brief increment in the intensity of a longer duration pedestal is commonly used as a measure of intensity-resolution. Increment detection is known to improve with increasing duration of the increment and also with increasing duration of the pedestal, but the relative effects of these two parameters have not been explored in the same study. In several past studies of the effects of increment duration, pedestal duration was increased as increment duration increased. In the present study, increment and pedestal duration were independently manipulated. Increment-detection thresholds were determined for four subjects with normal-hearing using a 500- or 4000-Hz pedestal presented at 60 dB sound pressure level (SPL). Increment durations were 10, 20, 40, 80, 160, and 320 ms. Pedestal durations were 20, 40, 80, 160, and 320 ms. Each increment duration was combined with all pedestals of equal or greater duration. Multiple-regression analyses indicate that increment detection under these conditions is determined primarily by pedestal duration. Follow-up experiments ruled out effects of off-frequency listening or overshoot. The results suggest that effects of increment duration have been confounded by effects of pedestal duration in studies that co-varied increment and pedestal duration. Implications for models of temporal integration are discussed.


Role of Rho-associated kinase in neointima formation after vascular injury.
The Rho/Rho-associated kinase (Rho-kinase) system is implicated in various cellular functions, including migration, proliferation, and apoptosis. Because a possible role of the system is suggested in neointima formation after vascular injury, we sought to examine whether a new specific Rho-kinase inhibitor, Y27632, prevents neointima formation of the balloon-injured rat carotid artery, and if so, to investigate the effects of Y27632 on migration, proliferation, and apoptosis of smooth muscle cells (SMCs) in the injured artery. Y27632 was administered intraperitoneally from 1 day before to 14 days after vascular injury. Treatment with Y27632 inhibited phenylephrine-induced Rho-kinase activation in the carotid artery on the basis of immunoblotting against the phosphorylated myosin-binding subunit of myosin phosphatase. Y27632 markedly prevented neointima formation at days 7 and 14. In controls, BrdU(+) proliferating and TUNEL(+) apoptotic SMCs were transiently and coincidentally increased in the neointima, with a peak at day 7. Y27632 significantly increased the neointimal TUNEL(+) SMCs at days 7 and 14, but not BrdU(+) SMCs. Y27642 significantly decreased the number of intimal SMCs at day 4, while not affecting the number of BrdU(+) or TUNEL(+) SMCs. Reendothelialization after balloon injury was not significantly affected by Y27632 at days 7 and 14. Y27632 inhibited neointima formation by enhancing SMC apoptosis and probably by suppressing early SMC migration. Therefore, a role of Rho-kinase is suggested in neointima formation after vascular injury.


Quantitative assessment of successive carbohydrate additions to the clustered O-glycosylation sites of IgA1 by glycosyltransferases.
Mucin-type O-glycosylation occurs on many proteins that transit the Golgi apparatus. These glycans impact structure and function of many proteins and have important roles in cellular biosynthetic processes, signaling and differentiation. Although recent technological advances have enhanced our ability to profile glycosylation of glycoproteins, limitations in the understanding of the biosynthesis of these glycan structures remain. Some of these limitations stem from the difficulty to track the biosynthetic process of mucin-type O-glycosylation, especially when glycans occur in dense clusters in repeat regions of proteins, such as the mucins or immunoglobulin A1 (IgA1). Here, we describe a series of nano-liquid chromatography (LC)-mass spectrometry (MS) analyses that demonstrate the range of glycosyltransferase enzymatic activities involved in the biosynthesis of clustered O-glycans on IgA1. By utilizing nano-LC-MS relative quantitation of in vitro reaction products, our results provide unique insights into the biosynthesis of clustered IgA1 O-glycans. We have developed a workflow to determine glycoform-specific apparent rates of a human UDP-N-acetylgalactosamine:polypeptide N-acetylgalactosaminyltrasnfersase (GalNAc-T EC 22.214.171.124) and demonstrated how pre-existing glycans affect subsequent activity of glycosyltransferases, such as core 1 galactosyltransferase and α2,3- and α2,6-specific sialyltransferases, in successive additions in the biosynthesis of clustered O-glycans. In the context of IgA1, these results have potential to provide insight into the molecular mechanisms implicated in the pathogenesis of IgA nephropathy, an autoimmune renal disease involving aberrant IgA1 O-glycosylation. In a broader sense, these methods and workflows are applicable to the studies of the concerted and competing functions of other glycosyltransferases that initiate and extend mucin-type core 1 clustered O-glycosylation.


[Selective reduction of the amplitude of the action potentials of neurons in the process of habituation].
Spike discharges of single neurons in the forebrain general cortex of the turtle were recorded extracellularly during habituation to light flashesmthe amplitude of spontaneous spikes, short-latent evoked spikes and spikes after the inhibitory pause was measuredmthe amplitude of short-latent evoked spikes significantly decreased during habtiuation while the background and postinhibitory spikes remained unchanged. It is supposed that the pattern of synapses activated by a definite stimulus may change the state of the elcetrically excitable neuronal membrane during habituation


Al Wakra emergency presentation of Behcet's disease.
This is atypical presentation of Behcet's disease and it's important for emergency physician and doctors who are working in acute medicine. We reported a case of a young patient presenting back pain concerning the right leg, a result of extensive bilateral deep vein thrombosis (DVT) in both legs that extended to inferior vena cava (IVC) and renal veins. After a bundle of investigations and medical examinations, the case was diagnosed as Behcet's disease with an atypical presentation. Al Wakra emergency presentation of Behcet's disease: low back pain with lower limb pain that result of DVT which is one of vascular complication of Behcet's disease.


Rapid identification of an adult plant stripe rust resistance gene in hexaploid wheat by high-throughput SNP array genotyping of pooled extremes.
High-throughput SNP array analysis of pooled extreme phenotypes in a segregating population by KASP marker genotyping permitted rapid, cost-effective location of a stripe rust resistance QTL in wheat. German wheat cultivar "Friedrichswerther" has exhibited high levels of adult plant resistance (APR) to stripe rust in field environments for many years. F2:3 lines and F6 recombinant inbred line (RILs) populations derived from a cross between Friedrichswerther and susceptible landrace Mingxian 169 were evaluated in the field in 2013, 2016 and 2017. Illumina 90K iSelect SNP arrays were used to genotype bulked extreme pools and parents; 286 of 1135 polymorphic SNPs were identified on chromosome 6B. Kompetitive Allele-Specific PCR (KASP) markers were used to verify the chromosome region associated with the resistance locus. A linkage map was constructed with 18 KASP-SNP markers, and a major effect QTL was identified within a 1.4 cM interval flanked by KASP markers IWB71602 and IWB55937 in the region 6BL3-0-0.36. The QTL, named QYr.nwafu-6BL, was stable across environments, and explained average 54.4 and 47.8% of the total phenotypic variation in F2:3 lines and F6 RILs, respectively. On the basis of marker genotypes, pedigree analysis and relative genetic distance QYr.nwafu-6BL is likely to be a new APR QTL. Combined high-throughput SNP array genotyping of pooled extremes and validation by KASP assays lowers sequencing costs compared to genome-wide association studies with SNP arrays, and more importantly, permits rapid isolation of major effect QTL in hexaploid wheat as well as improving accuracy of mapping in the QTL region. QYr.nwafu-6BL with flanking KASP markers developed and verified in a subset of 236 diverse lines can be used in marker-assisted selection to improve stripe rust resistance in breeding programs.


Airway inflammation in cystic fibrosis.
Patients with cystic fibrosis (CF) experience declining pulmonary function related to chronic airway inflammation, which results from epithelial and immune cell secretion of proinflammatory mediators that promote neutrophil influx into the airways. This inflammatory response may be disproportionate to the inciting infectious stimulus, resulting in an overly exuberant influx of neutrophils. The neutrophils release proteases, including neutrophil elastase, that eventually overwhelm the antiprotease capacity of the lung and cleave structural proteins, leading to bronchiectasis. This deleterious inflammatory process in patients with CF has become a potential therapeutic target, though the development of effective antiinflammatory therapies has been limited by the lack of sensitive outcome measures. Historically, indirect measures of lung disease, such as spirometry, have been used to assess the effect of antiinflammatory drugs. BAL remains the primary method of interrogating the inflammatory status of the airway, but the procedure is invasive and may eventually be supplanted by induced sputum. Anatomic imaging with high-resolution CT scanning is used clinically, but has unknown utility, and functional imaging, using positron emission tomography, appears promising but is still investigational. Despite the paucity of outcome measures, clinical trials of antiinflammatory agents, including corticosteroids and ibuprofen, have demonstrated benefits, though their use has been limited by adverse effects. Azithromycin is increasingly used as an immunomodulatory agent, although its mechanism of action remains unclear. Strategies for modulating the airway inflammation in patients with CF are currently under development and may offer new therapeutic options for these patients.


Demonstration of inter- and intracellular distribution of boron and gadolinium using micro-proton-induced X-ray emission (Micro-PIXE).
Micro-proton-induced X-ray emission (Micro-PIXE) was applied to determine inter- and intracellular distribution of boron (10B) and gadolinium (157Gd), the capture atoms used to kill tumor cells in neutron capture therapy (NCT). Cultured 9L gliosarcoma cells on Mylar film were exposed to sodium borocaptate (BSH) and gadobenate dimeglumine (Gd-BOPTA). To analyze the inter- and intracellular distribution of 10B and 157Gd in 9L gliosarcoma cells, the cells were irradiated using a proton beam of 1.7 or 3 MeV energy collimated to 1 microm diameter and emission X-ray was detected. The distribution of 10B and 157Gd in 9L gliosarcoma cells was then examined. In this study, we could directly analyze the inter- and intracellular distribution of 10B and 157Gd elements in 9L gliosarcoma cells directly using Micro-PIXE. This is the first report on the distribution of 10B employing a method to detect gamma-rays resulting from the nuclear reaction of 10B using particle-induced gamma-ray emission (PIGE). These results show that the distribution of 157Gd elements was correctly measured using micro-PIXE. 157Gd should have the same tendency as 10B in cultured 9L gliosarcoma cells and agree with the distribution in 9L gliosarcoma cells. Further investigation is necessary for a higher spatial resolution and optimization of the measurement time or improvement of the sampling method. In the future, it will be possible to employ this method to analyze the intracellular microdistribution of the capture element and in the development of new drugs for NCT.


Drosophila glypicans regulate the germline stem cell niche.
Stem cells are maintained in vivo by short-range signaling systems in specialized microenvironments called niches, but the molecular mechanisms controlling the physical space of the stem cell niche are poorly understood. In this study, we report that heparan sulfate (HS) proteoglycans (HSPGs) are essential regulators of the germline stem cell (GSC) niches in the Drosophila melanogaster gonads. GSCs were lost in both male and female gonads of mutants deficient for HS biosynthesis. dally, a Drosophila glypican, is expressed in the female GSC niche cells and is responsible for maintaining the GSC niche. Ectopic expression of dally in the ovary expanded the niche area, showing that dally is required for restriction of the GSC niche space. Interestingly, the other glypican, dally-like, plays a major role in regulating male GSC niche maintenance. We propose that HSPGs define the physical space of the niche by serving as trans coreceptors, mediating short-range signaling by secreted factors.


Update on the concepts and probable role of peptidergic nerves in dentine sensitivity and pain mechanisms.
Both, myelinated (A-) and unmyelinated (C-) and efferent sympathetic nerves are found in the dental pulp. The myelinated nerves are sensory and respond to stimulation of dentine with external stimuli. The A-fibres may be responsible for the sharp piercing pain sensations induced by stimulation of dentine in human subjects. C-fibres respond only when the stimuli reach the pulp proper. Activation of C-fibres may be responsible for the dull, delayed pain which radiates to the rest of the face. Immunohistochemical studies have demonstrated the existence of intradental peptidergic nerves. These are the substance P (SP), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP) nerves. It is suggested that peptidergic intradental nerves may play a role in increased dentine sensitivity during inflammation and dentine hypersensitivity.


Dynamics of human DNA topoisomerases IIalpha and IIbeta in living cells.
DNA topoisomerase (topo) II catalyses topological genomic changes essential for many DNA metabolic processes. It is also regarded as a structural component of the nuclear matrix in interphase and the mitotic chromosome scaffold. Mammals have two isoforms (alpha and beta) with similar properties in vitro. Here, we investigated their properties in living and proliferating cells, stably expressing biofluorescent chimera of the human isozymes. Topo IIalpha and IIbeta behaved similarly in interphase but differently in mitosis, where only topo IIalpha was chromosome associated to a major part. During interphase, both isozymes joined in nucleolar reassembly and accumulated in nucleoli, which seemed not to involve catalytic DNA turnover because treatment with teniposide (stabilizing covalent catalytic DNA intermediates of topo II) relocated the bulk of the enzymes from the nucleoli to nucleoplasmic granules. Photobleaching revealed that the entire complement of both isozymes was completely mobile and free to exchange between nuclear subcompartments in interphase. In chromosomes, topo IIalpha was also completely mobile and had a uniform distribution. However, hypotonic cell lysis triggered an axial pattern. These observations suggest that topo II is not an immobile, structural component of the chromosomal scaffold or the interphase karyoskeleton, but rather a dynamic interaction partner of such structures.


Spatial orientation in humans: perception of angular whole-body displacements in two-dimensional trajectories.
Vestibular perception of whole-body passive rotation in the horizontal plane was studied by applying two-dimensional (2D) motion to eight blindfolded healthy volunteers: pure rotations in place, corner-like trajectories and arcs of a circular trajectory were randomly applied by means of a remotely controlled robot. Angles embedded in the 2D trajectories were 45 degrees, 90 degrees, 135 degrees and 180 degrees. Stimulation of semicircular canals was the same for all trajectories but was accompanied by concurrent otolith stimulation during circular motion. Subjects participated in two successive experimental sessions. In the first session they were instructed to use a pointer to reproduce the total angular displacement after the motion (REPRODUCTION); in the second session they had to keep pointing towards a remote (15 m) memorised target during the motion (TRACKING). In REPRODUCTION subjects tended to overestimate their rotation angle by 28 +/- 11% (mean +/- SD). There was no systematic effect of the trajectory. Overestimation also occurred when subjects were required to rotate in darkness by 180 degrees (by controlling a joystick). In TRACKING there was virtually no overestimation (6 +/- 17%) and the movement of the pointer matched the dynamics of angular motion. We conclude that (a) the brain can separate and memorise the angular component of complex 2D motion; however, a large inter-individual variability in estimating its amplitude exists; (b) in the range of linear accelerations used in the study, no appreciable effect of otolith-canal perceptual interaction was shown; (c) angular displacements can be dynamically transformed into matched pointing movements; (d) overestimation seems to be typical of delayed judgements of angular displacement and of self-controlled rotations in place. This could be due to the characteristics of the physiological calibration of the vestibular input.


Photosynthetic Response of Soybean Leaf to Wide Light-Fluctuation in Maize-Soybean Intercropping System.
In maize-soybean intercropping system, soybean plants will be affected by the wide light-fluctuation, which resulted from the shading by maize plants, as the shading of maize the light is not enough for soybean in the early morning and late afternoon, but at noon, the light is strong as the maize shading disappeared. The objective of this study is to evaluate the photosynthetic response of soybean leaf to the wide light-fluctuation. The data of diurnal variation of photosynthetic characters showed that the photosynthetic rate of intercropped soybean was weaker than that of monocropped soybean. The chlorophyll content, ratio of chlorophyll a/b, and AQE (apparent quantum efficiency) were increased and Rd (dark respiration rate) was decreased for the more efficient interception and absorption of light and carbon gain in intercropping. δRo (The efficiency/probability with which an electron from the intersystem electron carriers was transferred to reduce end electron acceptors at the PSI acceptor side) and φRo (the quantum yield for the reduction of the end electron acceptors at the PSI acceptor side) in intercropped soybean leaf were lower compared to those in monocropped one, which showed that the acceptor side of PSI might be inhibited, and also it was the main reason that soybean plants showed a low photosynthetic capacity in intercropping. ψEo (the efficiency/probability with an electron moves further than QA-) in monocropping and intercropping decreased 5.8, and 35.7%, respectively, while φEo (quantum yield for electron transport) decreased 27.7 and 45.3% under the high radiation at noon, which suggested that the acceptor side of PSII was inhibited, while the NPQ became higher. These were beneficial to dissipate excess excitation energy in time, and protect the photosynthetic apparatus against photo-damage. The higher performance index on the absorption basis (PIABS) and lower δRo, φRo, ψEo, and φEo of intercropped soybeans compared to monocropping under high radiation indicated that the electron transfer of intercropped soybean was inhibited more seriously and intercropped soybean adjusted the electron transport between PSII to PSI to adapt the light-fluctuation. Higher NPQ capacity of intercropped soybeans played a key role in keeping the leaf with a better physiological flexibility under the high radiation.