Serodiagnosis of equine leptospirosis by enzyme-linked immunosorbent assay using four recombinant protein markers.
Leptospirosis, caused by Leptospira spp., is one of the most common zoonotic diseases in the world. We tested four recombinant proteins of Leptospira interrogans, namely, rLipL21, rLoa22, rLipL32, and rLigACon4-8, to evaluate their potential for use as antigens for the diagnosis of equine leptospirosis. We employed equine sera (n = 130) that were microscopic agglutination test (MAT) negative and sera (n = 176) that were MAT positive for the 5 serovars that most commonly cause equine leptospirosis. The sensitivity and specificity of ELISA compared to MAT were 82.39% and 86.15%, respectively, for LigACon4-8, 77.84% and 92.31%, respectively, for Loa22, 77.84% and 86.15%, respectively, for LipL32, and 84.66% and 83.85%, respectively, for LipL21. When one of the two antigens was test positive, the sensitivity and specificity of ELISA were 93.75% and 78.46%, respectively, for rLigACon4-8 and LipL32, 93.18% and 76.15%, respectively, for rLigACon4-8 and LipL21, 89.77% and 80.77%, respectively, for rLigACon4-8 and Loa22, 91.48% and 78.46%, respectively, for LipL21 and Loa22, 93.75% and 76.92%, respectively, for LipL21 and LipL32, and 90.34% and 80.77%, respectively, for Loa22 and LipL32. In conclusion, we have developed an indirect ELISA utilizing rLigACon4-8, rLoa22, rLipL32, and rLipL21 as diagnostic antigens for equine leptospirosis. The use of four antigens in the ELISA was found to be sensitive and specific, the assay was easy to perform, and the results concurred with the results of the standard Leptospira MAT.


Pharmacogenomic profiling and pathway analyses identify MAPK-dependent migration as an acute response to SN38 in p53 null and p53-mutant colorectal cancer cells.
The topoisomerase I inhibitor irinotecan is used to treat advanced colorectal cancer and has been shown to have p53-independent anticancer activity. The aim of this study was to identify the p53-independent signaling mechanisms activated by irinotecan. Transcriptional profiling of isogenic HCT116 p53 wild-type and p53 null cells was carried out following treatment with the active metabolite of irinotecan, SN38. Unsupervised analysis methods showed that p53 status had a highly significant impact on gene expression changes in response to SN38. Pathway analysis indicated that pathways involved in cell motility [adherens junction, focal adhesion, mitogen-activated protein kinase (MAPK), and regulation of the actin cytoskeleton] were significantly activated in p53 null cells, but not p53 wild-type cells, following SN38 treatment. In functional assays, SN38 treatment increased the migratory potential of p53 null and p53-mutant colorectal cancer cell lines, but not p53 wild-type lines. Moreover, p53 null SN38-resistant cells were found to migrate at a faster rate than parental drug-sensitive p53 null cells, whereas p53 wild-type SN38-resistant cells failed to migrate. Notably, cotreatment with inhibitors of the MAPK pathway inhibited the increased migration observed following SN38 treatment in p53 null and p53-mutant cells. Thus, in the absence of wild-type p53, SN38 promotes migration of colorectal cancer cells, and inhibiting MAPK blocks this potentially prometastatic adaptive response to this anticancer drug.


Association studies in outbred mice in a new era of full-genome sequencing.
Thousands of loci that contribute to quantitative traits in outbred crosses of mice have been reported over the last two decades. In this review we discuss how outbred mouse populations can be used to map and identify the genes and sequence variants that give rise to quantitative variation. We discuss heterogeneous stocks, the diversity outbred, and commercially available outbred populations of mice. All of these populations are descended from a small number of progenitor strains. The availability of the complete sequence of laboratory strains means that in many cases it will be possible to reconstruct the genomes of the outbred animals so that in a genetic association study we can detect the effect of all variants, a situation that has so far eluded studies in completely outbred populations. These resources constitute a major advance and make it possible to progress from a quantitative trait locus to a gene at an unprecedented speed.


Primary health care in Canada: in praise of the nurse?
In Canada, as elsewhere, it has been agreed that primary health care (PHC) is needed, particularly for the underserved population. But the debate continues over which approach, what measures and who should be involved. To help find the answers, the Canadian Nurses Association presented a working paper already in 1980 to the Canadian Government recommending changes in the health system and proposing various roles that nurses could play in formulating and implementing its national primary health policy and in November this year organized a two-day PHC workshop involving representatives from provincial councils and government as well as nurses. Below, two nurses teaching at the University of Ottawa's School of Nursing give their view on what should and could be done in Canada.


NMR analysis of an Fe(i)-carbene complex with strong magnetic anisotropy.
A tricoordinated FeI complex with two cyclic-alkyl(amino) carbene (cAAC) and one chlorido ligand, (cAAC)2FeCl (1), is studied by means of 1H NMR spectroscopy and DFT calculations. Due to the cAAC ligands, which can take significant amounts of spin density from the metal center, and due to the magnetic anisotropy of the FeI ion (P. P. Samuel et al., J. Am. Chem. Soc., 2014, 136, 11964-11971), compound 1 is a rare example of a paramagnetic d-block compound which is expected to have significant contributions from both contact and pseudocontact terms to the hyperfine NMR shift. Compound 1 is fluxional, which makes the analysis of its 1H NMR spectrum more difficult but allows a preliminary assignment from EXSY spectra. Then, a software-aided approach enabled a satisfactory signal assignment of all protons which are distanced from the FeI center and carbene cyclic core, and thereby the extraction of the axial and rhombic components of the magnetic susceptibility anisotropy tensor (Δχ). Components of Δχ enable the calculation of zero-field spitting D and E parameters from solution NMR measurements of 1, and these parameters are compared to previously reported experimental and theoretical values.


The Effect of Gluten-free Diet on Cardiovascular Risk Factors in Newly Diagnosed Pediatric Celiac Disease Patients.
Although gluten-free diet (GFD) is the only proven therapy for celiac disease (CD), its effect on cardiovascular disease (CVD) risk factors is still unclear. Our aim was to determine whether adherence to GFD affects CVD risk factors among newly diagnosed pediatric CD subjects. We prospectively enrolled pediatric subjects undergoing upper gastrointestinal endoscopy for suspected CD. We collected anthropometric and laboratory parameters related to CVD risk factors at the time of CD diagnosis and 1 year after initiation of a GFD and evaluated changes in CVD risk factors. Paired t tests or Wilcoxon nonparametric tests were used, each when appropriate. One hundred ten newly diagnosed CD pediatric subjects were included in the analysis. There were 64 (58.2%) girls and the mean age at diagnosis was 6.8 ± 3.4 years. Median body mass index z scores (P = 0.84), rates of underweight or overweight (P = 0.32), and rates of elevated blood pressure (P = 0.78) remained unchanged. Although median fasting insulin levels increased (1.9 vs 5.4 μU/mL, P < 0.001), insulin resistance as measured by homeostatic model assessment did not increase after 1 year of GFD (P = 0.16). Although rates of dyslipidemia remained unchanged, median high-density lipoprotein levels increased on GFD (47 vs 51 mg/dL, P < 0.001). In this pediatric CD cohort, GFD for 1 year was not associated with increased CVD risk factors. The long-term significance of these mild changes is yet to be determined.


Lynch Syndrome and Endometrial Cancer.
In patients with Lynch syndrome, gynecologic cancer often can be the first presenting malignancy. In this review, we summarize the genetics of Lynch syndrome and review the various modalities of identifying patients at risk for this syndrome. The clinical characteristics of Lynch-associated endometrial cancer and screening and risk-reducing strategies also are described.


Multi-drug-resistant Staphylococcus aureus and future chemotherapy.
Staphylococcus (S.) aureus silently stays as our natural flora, and yet sometimes threatens our life as a tenacious pathogen. In addition to its ability to outwit our immune system, its multi-drug resistance phenotype makes it one of the most intractable pathogenic bacteria in the history of antibiotic chemotherapy. It conquered practically all the antibiotics that have been developed since 1940s. In 1961, the first MRSA was found among S. aureus clinical isolates. Then MRSA prevailed throughout the world as a multi-resistant hospital pathogen. In 1997, MRSA strain Mu50 with reduced susceptibility to vancomycin was isolated. Vancomycin-intermediate S. aureus (VISA), so named according to the CLSI criteria, was the product of adaptive mutation of S. aureus against vancomycin that had long been the last resort to MRSA infection. Here, we describe the genetic basis for the remarkable ability of S. aureus to acquire multi-antibiotic resistance, and propose a novel paradigm for future chemotherapy against the multi-resistant pathogens.


Mouse respiratory epithelial cells support efficient replication of human rhinovirus.
Human rhinoviruses (HRV) are responsible for the majority of virus infections of the upper respiratory tract. Furthermore, HRV infection is associated with acute exacerbation of asthma and other chronic respiratory diseases of the lower respiratory tract. A small animal model of HRV-induced disease is required for the development of new therapies. However, existing mouse models of HRV infection are difficult to work with and until recently mouse cell lines were thought to be generally non-permissive for HRV replication in vitro. In this report we demonstrate that a virus of the minor receptor group, HRV1B, can infect and replicate in a mouse respiratory epithelial cell line (LA-4) more efficiently than in a mouse fibroblast cell line (L). The major receptor group virus HRV16 requires human intercellular adhesion molecule-1 (ICAM-1) for cell entry and therefore cannot infect LA-4 cells. However, transfection of in vitro-transcribed HRV16 RNA resulted in the replication of viral RNA and production of infectious virus. Expression of a chimeric ICAM-1 molecule, comprising mouse ICAM-1 with extracellular domains 1 and 2 replaced by the equivalent human domains, rendered the otherwise non-permissive mouse respiratory epithelial cell line susceptible to entry and efficient replication of HRV16. These observations suggest that the development of mouse models of respiratory tract infection by major as well as minor group HRV should be pursued.


SETDB1 regulates SMAD7 expression for breast cancer metastasis.
Breast cancer (BRC) is the most invasive cancer in women. Although the survival rate of BRC is gradually increasing due to improved screening systems, development of novel therapeutic targets for inhibition of BRC proliferation, metastasis and recurrence have been constantly needed. Thus, in this study, we identified overexpression of SETDB1 (SET Domain Bifurcated 1), a histone methyltransferase, in RNA-seq data of BRC derived from TCGA portal. In Gene Ontology (GO) analysis, cell migration-related GO terms were enriched, and we confirmed down-regulation of cell migration/invasion and alteration of EMT /MET markers after knockdown of SETDB1. Moreover, gene network analysis showed that SMAD7 expression is regulated by SETDB1 levels, indicating that up-regulation of SMAD7 by SETDB1 knockdown inhibited BRC metastasis. Therefore, development of SETDB1 inhibitors and functional studies may help develop more effective clinical guidelines for BRC treatment. [BMB Reports 2019; 52(2): 139-144].


A computational and functional study elicits the ameliorating effect of the Chinese herbal formula Huo Luo Xiao Ling Dan on experimental ischemia-induced myocardial injury in rats via inhibition of apoptosis.
Ischemic heart disease (IHD) is the leading cause of death worldwide and remains a major life-threatening factor in humans. Apoptosis has been implicated in the pathogenesis of IHD. The Chinese herbal formula Huo Luo Xiao Ling Dan (HLXLD), one of the commonly used Chinese herbal formulas, consists of Salviae miltiorrhizae, Angelica sinensis, Gummi olibanum, and Commiphora myrrha, with a wide spectrum of pharmacological activity. However, the mechanism of action and molecular targets of HLXLD in the treatment of IHD are unclear. This study aimed to computationally predict the molecular interactions between the major active components of HLXLD and key regulators of apoptosis and then examine the effect of HLXLD on coronary artery ligation-induced acute myocardial ischemia in rats. The molecular interactions between the major active components of HLXLD, including ferulic acid, ligustilide, succinic acid, vanillic acid, tanshinone IIA, tanshinone IIB, danshensu, salvianolic acid A, salvianolic acid C, protocatechuic aldehyde, and β-boswellic acid and human protein molecules including B cell lymphoma-extra large (Bcl-xl), B cell lymphoma 2 antagonist/killer 1 (Bak1), B cell lymphoma 2 (Bcl-2), procaspase 3, and caspase 9 with regard to hydrogen bond formation, charge interaction, and π-π stacking using Discovery Studio(®) program 3.1. The 12 HLXLD components were predicted by ADMET (absorption, distribution, metabolism, excretion and toxicity) Predictor to have favorable pharmacokinetic and low hepatotoxicity profiles. The acute myocardial ischemia was established by surgical ligation of the left anterior descending coronary artery. The rats were divided into a sham operative group, a model group, a positive control group treated with 0.2 mg/kg isosorbide mononitrate, and groups treated with 2.7, 5.4, or 10.8 g/kg HLXLD. The results showed that administration of HLXLD increased mean arterial pressure, left ventricular systolic pressure, heart rate, and maximal rate of rise/descent of left ventricular pressure levels. Administration of HLXLD significantly ameliorated coronary artery ligation-induced tissue damage in the left ventricle, with restored arrangement of myocardial fibers and recovered myoplasm in rats. Furthermore, HLXLD markedly increased the expression level of Bcl-2 but decreased the level of cleaved caspase 3. Taken together, administration of HLXLD attenuated acute myocardial ischemia-induced damage in cardiomyocytes and inhibited apoptotic death of cardiomyocytes, thereby exerting a cardioprotective effect in rats with IHD. These findings suggest that HLXLD may represent a promising herbal formula for the treatment of cardiovascular disease by counteracting apoptotic cell death via multiple active compounds. More studies are warranted to fully elucidate the mechanisms of action, identify the therapeutic targets, and validate the efficacy and safety of HLXLD in the treatment of IHD.


Influence of drought and total phosphorus on diel pH in wadeable streams: implications for ecological risk assessment of ionizable contaminants.
Climatological influences on site-specific ecohydrology are particularly germane in semiarid regions where instream flows are strongly influenced by effluent discharges. Because many traditional and emerging aquatic contaminants, such as pharmaceuticals, are ionizable, we examined diel surface water pH patterns (i.e., change in pH over a 24-h period) at 23 wadeable streams in central Texas, USA, representing a gradient of nutrient enrichment during consecutive summers of 2006 and 2007. The years of our study were characterized by decidedly different instream flows, which likely affected production:respiration dynamics and led to distinctions in diel pH patterns between 2006 and 2007. Site-specific ambient water quality criteria for NH(3) and the aquatic toxicity of the model weak base pharmaceutical sertraline were predicted using continuous water quality monitoring data from the sites. Drought conditions of 2006 significantly increased (p<0.05) diel pH changes compared to high instream flows of 2007,and the magnitude of diel pH variability was most pronounced at nutrient-enriched sites in 2006. Differences in diel pH change patterns between 2006 and 2007 affected predictions of the environmental fate and effects for model weak base pharmaceuticals and NH(3). Overall, site-specific diel pH was more variable at some sites than the difference in mean surface water pH between the 2 summers. Diel pH variability affected regulatory criteria, because 20% of the study sites in 2006 experienced greater than 5-fold differences in National Ambient Water Quality Criteria for NH(3) over 24-h periods. Our study emphasizes the potential uncertainty that diel pH variability may introduce in site-specific assessments and provides recommendations for environmental assessment of ionizable contaminants.


The prevalence of Gasterophilus intestinalis (Diptera: Oestridae) in donkeys (Equus asinus) in Egypt with special reference to larvicidal effects of neem seed oil extract (Azadirachta indica) on third stage larvae.
Gasterophiline larvae are of veterinary and medical importance caused specific equine intestinal myiasis. Gasterophilus intestinalis (Botfly larvae) had a wide geographical distribution. The present study explores the prevalence rate of G. intestinalis 3rd stage larvae in Egypt from January- December 2017; besides, in vitro trials to control of this larvae and evaluation of this trial using Scanning Electron Microscope (SEM) and histopathology of treated larvae. In the present study, the 3rd larval stage of G. intestinalis was found in clusters in the epithelium of the investigated stomach and infested with prevalence rate 97.2%. The highest collected numbers of larvae were found in two months; March and August (570 & 520 larvae) and lowest numbers (200 larvae) were collected in October, November, and December. The calculated LC50 and LC90 values of neem seed extract were 707.9 ppm and 1090.7 ppm. The different alteration was recorded after exposure to oil extract which showed some destruction on cuticle surface as folded and corrugated cuticle, destruction of maxillae with pits on its surface, disfigure and irregularity of cephalic spines. Histopathology of exposed G. intestinalis larvae showed different changes as thinning of cuticle at the different level (exocuticle, endocuticle, cell layers), degeneration of epithelial cells of the gut and different degree of necrosis was described. Life cycle of G.intestinalis was followed up after treatment with neem seed extract.


Osmotic and ionic regulation in different populations of the new zealand freshwater crayfish Paranephrops zealandicus.
1. Some features of the osmoregulatory mechanism are compared in four populations of Paranephrops zeal andicus White collected form freshwaters of different ionic concentrations. 2. Crayfish from freshwaters of ca, 2-0 mM-NaCl concentration show a sustained decrease in blood concentration of ca. 8% when placed in 0-2 mM-NaCl. 3. Populations from freshwaters of ca. 0-2-0-4 mM-NaCl show lower rates of net salt loss in distilled water and higher rates of net salt uptake form dilute NaCl solutions than do populations from freshwaters of ca. 0-8-2-0 mM-NaCl. 4. Renal salt losses over the first 24 h in distilled water account for ca. 18% of the total salt loss. 5. It is suggested that P. zealandicus from environments of lowest concentration shows a similar degree of adaptation to freshwater as do crayfish of the northern hemisphere. It differs in possessing a substantially higher blood concentration.


Differential actigraphy for monitoring asymmetry in upper limb motor activities.
Most applications of accelerometry-based actigraphy require a single sensor, properly located onto the body, to estimate, for example, the level of activity or the energy expenditure. Some approaches adopt a multi-sensor setup to improve those analyses or to classify different types of activity. The specific case of two symmetrically placed actigraphs allowing, by some kind of differential analysis, for the assessment of asymmetric motor behaviors, has been considered in relatively few studies. This article presents a novel method for differential actigraphy, which requires the synchronized measurements of two triaxial accelerometers (programmable eZ430-Chronos, Texas Instruments, USA) placed symmetrically on both wrists. The method involved the definition of a robust epoch-related activity index and its implementation on-board the adopted programmable platform. Finally, the activity recordings from both sensors allowed us to define a novel asymmetry index AR24 h ranging from -100% (only the left arm moves) to +100% (only the right arm moves) with null value marking a perfect symmetrical behavior. The accuracy of the AR24 h index was 1.3%. Round-the-clock monitoring on 31 healthy participants (20-79 years old, 10 left handed) provided for the AR24 h reference data (range -5% to 21%) and a fairly good correlation to the clinical handedness index (r = 0.66, p < 0.001). A subset of 20 participants repeated the monitoring one week apart evidencing an excellent test-retest reliability (r = 0.70, p < 0.001). Such figures support future applications of the methodology for the study of pathologies involving motor asymmetries, such as in patients with motor hemisyndromes and, in general, for those subjects for whom a quantification of the asymmetry in daily motor performances is required to complement laboratory tests.


Low-salt O-miso produced from Koji fermentation of oncom improves redox state and cholesterolemia in rats more than low-salt soybean-miso.
The author prepared low-salt miso by koji fermentation using soy-oncom and okara-oncom (9:1), at which time soybeans and okara were fermented with Neurospora intermedia (oncom miso, i.e. O-miso). Its usefulness as a seasoning, as well as high antioxidative activity in vitro and antimutagenicity have already been presented. In this study, the antioxidative activity of O-miso in vivo, as well as serum cholesterol-lowering action, were investigated concerning contribution to health. In rats fed O-miso, the serum alpha-tocopherol level and isoflavone-aglycone-intake were higher than in rats fed low-salt soybean miso (S-miso). The activities of serum GSH-Px and hepatic catalase were also higher in rats fed O-miso than in rats fed S-miso, while the TBARS values were lower in both the serum and livers of rats fed O-miso. Furthermore, O-miso intake suppressed a rise in serum cholesterol level and increased fecal bile acid excretion. Such a cholesterol-lowering action of O-miso may be attributable to the micelle-breaking or evacuant effects of indigestible proteinous residues and the antioxidative activity of isoflavone-aglycones. O-miso seems to be more beneficial to health than S-miso in view of these aspects.


Potential Drug Interactions of Repurposed COVID-19 Drugs with Lung Cancer Pharmacotherapies.
Lung cancer patients are at heightened risk for developing COVID-19 infection as well as complications due to multiple risk factors such as underlying malignancy, anti-cancer treatment induced immunosuppression, additional comorbidities and history of smoking. Recent literatures have reported a significant proportion of lung cancer patients coinfected with COVID-19. Chloroquine, hydroxychloroquine, lopinavir/ritonavir, ribavirin, oseltamivir, remdesivir, favipiravir, and umifenovir represent the major repurposed drugs used as potential experimental agents for COVID-19 whereas azithromycin, dexamethasone, tocilizumab, sarilumab, famotidine and ceftriaxone are some of the supporting agents that are under investigation for COVID-19 management. The rationale of this review is to identify potential drug-drug interactions (DDIs) occurring in lung cancer patients receiving lung cancer medications and repurposed COVID-19 drugs using Micromedex and additional literatures. This review has identified several potential DDIs that could occur with the concomitant treatments of COVID-19 repurposed drugs and lung cancer medications. This information may be utilized by the healthcare professionals for screening and identifying potential DDIs with adverse outcomes, based on their severity and documentation levels and consequently design prophylactic and management strategies for their prevention. Identification, reporting and management of DDIs and dissemination of related information should be a major consideration in the delivery of lung cancer care during this ongoing COVID-19 pandemic for better patient outcomes and updating guidelines for safer prescribing practices in this coinfected condition.


[COVID-19 andcardiovascular diseases].
COVID-2019 disease mainly affects the respiratory tract and can progress in severe cases to pneumonia, acute respiratory distress syndrome and multi-organ failure. Patients with prior cardiovascular disease are at higher risk of developing an infection and progressing to a severe form of the disease. Also, due to the growing number of infected cases, it is clear that, in addition to the typical respiratory symptoms caused by the infection, some patients suffer from cardiovascular damage. This condition can, in fact, cause significant myocardial damage, which worsens the disease and affects the prognosis. Based on the results of currently published research, it seems important to discuss the manifestations and characteristics of myocardial damage induced by COVID-19 and its impact on patient prognosis.


Risky business: focus-group analysis of sexual behaviors, drug use and victimization among incarcerated women in St. Louis.
Incarcerated women report multiple vulnerabilities and, yet, are under-represented in research. This study used focus-group methodology to explore high-risk sexual behaviors, drug use, and victimization among female offenders in St. Louis. Inmates of the St. Louis Medium Security Institution (MSI) were invited to participate in one of five focus groups between May and September 2005 in preparation for an NIH/NINR HIV-prevention intervention study among female offenders in Drug Court. The focus group sample of 30 women was 70% African-American, with a mean age of 36 years. Results indicated that oral sex was the most common sex trade activity. Consistent with the literature, condom usage was described as irregular. In terms of drug use, participants reported that crack was most commonly used, with binges often lasting for several days. Regarding victimization, women frequently reported sexual abuse in childhood, and some described abusive relationships as adults. Participants also reported being beaten and raped by customers, which led to their concealing knives in purses and razors under the tongue. Consequently, perpetrated violence, including murder, was reported as protection against further violence. These findings confirm the vulnerability of this population of women who are at high risk for HIV. Effective HIV-prevention interventions are needed to assist these incarcerated women in making lifestyle changes during incarceration and sustaining them after release.


Portal vein thrombosis.
Portal vein thrombosis is a condition not infrequently encountered by clinicians. It results from a combination of local and systemic prothrombotic risk factors. The presentation of acute thrombosis varies widely from an asymptomatic state to presence of life-threatening intestinal ischemia and infarction. In the chronic stage, patients typically present with variceal bleeding or other complications of portal hypertension. Abdominal ultrasound color Doppler imaging has a 98% negative predictive value, and is considered the imaging modality of choice in diagnosing portal vein thrombosis. Controlled clinical trials to assist with clinical decision-making are lacking in both acute and chronic portal vein thrombosis. Oral anticoagulant therapy is initiated if the risks of bleeding are low, but long-term anticoagulation is generally not recommended in patients with concomitant hepatic cirrhosis. The roles of invasive therapeutic approaches such as thrombolysis and transjugular intrahepatic portosystemic shunt continue to evolve. This review conflates dissenting views into a rational approach of managing patients with portal vein thrombosis for the general internist.


Light-controlled release of nitric oxide from solid polymer composite materials using visible and near infra-red light.
Photochemical Nitric oxide releasing composite materials (Photo-NORMs) were prepared using biocompatible polymers and the photochemical nitric oxide donor complex (CrONO). We have demonstrated nitric oxide (NO) release from the solid composites for extended (>30 hours) and controlled (20-100 pmoles s(-1)) durations after visible light irradiation. Quantitation of the efficiency of NO release from the composites shows that polymer gas permeability most dramatically affects the overall efficiency (QY) of photochemical NO release, where polymers with higher gas permeability have a higher QY of nitric oxide release. Composites were also prepared with β-phase lanthanide-doped NaYF4 upconverting nanoparticles (UCNPs). Controlled Nitric oxide release was achieved via near infrared (NIR) irradiation. A prototype LED device shows proof-of-concept that such photoresponsive NO-releasing composites could be applied to implantable systems, where the amount of NO released is modulated by changing irradiation time and light intensity. This research provides the guidelines necessary to move towards device fabrication and testing in actual tissue to evaluate the photo-NORMS as a reliable option for nitric oxide release in vivo.


Cysteine substitution of transmembrane domain amino acids alters the ethanol inhibition of GluN1/GluN2A N-methyl-D-aspartate receptors.
N-Methyl-d-aspartate receptors (NMDARs) are inhibited by behaviorally relevant concentrations of ethanol, and residues within transmembrane (TM) domains of NMDARs, including TM3 GluN1 phenylalanine 639 (F639), regulate this sensitivity. In the present study, we used cysteine (C) mutagenesis to determine whether there are additional residues within nearby TM domains that regulate ethanol inhibition on NMDARs. GluN1(F639C)/GluN2A receptors were less inhibited by ethanol than wild-type receptors, and inhibition was restored to wild-type levels following treatment with ethanol-like methanethiosulfonate reagents. Molecular modeling identified six residues in the GluN1 TM1 domain (valine V566; serine S569) and the GluN2A TM4 domain (methionine, M817; V820, F821, and leucine, L824) that were in close vicinity to the TM3 F639 residue, and these were individually mutated to cysteine and tested for ethanol inhibition and receptor function. The F639C-induced decrease in ethanol inhibition was blunted by coexpression of GluN1 TM1 mutants V566C and S569C, and statistically significant interactions were observed for ethanol inhibition among V566C, F639C, and GluN2A TM4 mutants V820C and F821C and S569C, F639C, and GluN2A TM4 mutants F821C and L824C. Ethanol inhibition was also reduced when either GluN1 TM1 mutant V566C or S569C was combined with GluN2A V820C, suggesting a novel TM1:TM4 intrasubunit site of action for ethanol. Cysteines substituted at TM3 and TM4 sites previously suggested to interact with ethanol had less dramatic effects on ethanol inhibition. Overall, the results from these studies suggest that interactions among TM1, TM3, and TM4 amino acids in NMDARs are important determinants of ethanol action at these receptors.


The relationship between echocardiographic parameters and albumin bilirubin score in patients with acute pulmonary thromboembolism.
The Albumin-Bilirubin (ALBI) score is useful and easy-to-use for objectively assessing liver function. We investigated whether the ALBI score, a parameter indicating liver stiffness, congestion and fibrosis, has any relationship with echocardiographic parameters in patients with acute pulmonary thromboembolism (PTE). A total of 140 patients diagnosed with acute PTE were retrospectively analyzed. These patients were divided into three groups according to the hemodynamic severity of acute PTE: Group I [Low risk]; Group II [Submassive or intermediate-risk]; and Group III [Massive or high-risk]. Biochemical data obtained from venous blood samples taken at admission were analyzed. In addition, data were also analyzed from transthoracic echocardiography and pulmonary computed tomographic angiography performed at admission. ALBI, Bova, and PESI scores were calculated. ALBI scores (-3.32 ± 0.21 vs -2.86 ± 0.15 vs -2.46 ± 0.2, p < .001) were statistically significantly higher in Group III than Groups I and II. There was a significant difference between the three groups in terms of echocardiographic parameters, and LVEF and TAPSE values tended to decrease from group I to group III. In multivariate linear regression analysis, sPAP, RV/RA diameter, and NT-pro-BNP were found to be significantly associated with the ALBI score. An ALBI score higher than -2.87 was associated with Bova stage II-III in patients with Group I and Group II PTE, with a sensitivity of 87% and a specificity of 62% (AUC = 0.804; 95% CI 0.713-0.895; p < .001). The ALBI score, which is a common, easy-to-use, and inexpensive method, may be beneficial to select intermediate and high-risk patients in patients with acute PTE. Additionally, it may have prognostic value in distinguishing low and intermediate-risk acute PTE patients.


Mathematical artificial intelligence design of mutation-proof COVID-19 monoclonal antibodies.
Emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants have compromised existing vaccines and posed a grand challenge to coronavirus disease 2019 (COVID-19) prevention, control, and global economic recovery. For COVID-19 patients, one of the most effective COVID-19 medications is monoclonal antibody (mAb) therapies. The United States Food and Drug Administration (U.S. FDA) has given the emergency use authorization (EUA) to a few mAbs, including those from Regeneron, Eli Elly, etc. However, they are also undermined by SARS-CoV-2 mutations. It is imperative to develop effective mutation-proof mAbs for treating COVID-19 patients infected by all emerging variants and/or the original SARS-CoV-2. We carry out a deep mutational scanning to present the blueprint of such mAbs using algebraic topology and artificial intelligence (AI). To reduce the risk of clinical trial-related failure, we select five mAbs either with FDA EUA or in clinical trials as our starting point. We demonstrate that topological AI-designed mAbs are effective to variants of concerns and variants of interest designated by the World Health Organization (WHO), as well as the original SARS-CoV-2. Our topological AI methodologies have been validated by tens of thousands of deep mutational data and their predictions have been confirmed by results from tens of experimental laboratories and population-level statistics of genome isolates from hundreds of thousands of patients.


Hyperbaric medicine simulation education curriculum: CNS toxicity during Treatment Table 6 and intubated ICU patient with mucous plugging.
The incidence of complications and number of critically ill patients in hyperbaric medicine is relatively low [1]. This poses a challenge to those tasked with educating trainees as well as maintaining the skills of staff. Hyperbaric medicine fellows may not be exposed to critical patient scenarios or complications of hyperbaric medicine during a one-year fellowship. Additional staff may be unfamiliar with these situations as well. The purpose of hyperbaric simulation curriculum is to train health care providers for rare situations. To our knowledge, this hyperbaric simulation curriculum is the first published use of simulation education in the specialty of undersea and hyperbaric medicine. Two simulation cases have been developed that involve a patient with oxygen toxicity during hyperbaric treatment as well as an ICU patient with mucous plugging. Medical training simulations are an effective method of teaching content and training multiple roles in Undersea and Hyperbaric Medicine. A hyperbaric simulation curriculum is an achievable educational initiative that is able to train multiple team members simultaneously in situations that they may not encounter on a regular basis. We believe that this could be easily exported to otherinstitutions for further education.


L-edge XANES analysis of photoexcited metal complexes in solution.
Ultrafast X-ray absorption spectroscopy is a powerful tool to observe electronic and geometric structures of short-lived reaction intermediates. The ab initio FEFF9 code is applied to simulate the Pt L(3)-edge XANES spectrum of the photocatalytic diplatinum molecule [Pt(2)(P(2)O(5)H(2))(4)](4-) and the photo-induced changes that occur therein. The spectra are interpreted within a XAFS-like scattering theoretical framework (bound-continuum transitions) or in terms of a final-state local l-projected density of states (LDOS) (bound-bound transitions). By using a novel Bayesian fitting procedure, we show that the ground-state structures obtained independently from the XANES and EXAFS regions of the spectrum are in good agreement with each other. The semi-quantitative result obtained for the Pt-Pt contraction in the excited state is in line with recently published values. The improved theoretical treatment of inelastic losses has shown to result in more accurate peak positions in the above-continuum region of the spectrum which is an important prerequisite for obtaining quantitative structural information from (time-resolved) XANES spectra.


Intrathymic expression of Flt3 ligand enhances thymic recovery after irradiation.
Hematopoietic stem cell transplantation (HSCT) requires conditioning treatments such as irradiation, which leads to a severely delayed recovery of T cell immunity and constitutes a major complication of this therapy. Currently, our understanding of the mechanisms regulating thymic recovery is limited. It is known that a subpopulation of bone marrow (BM)-derived thymic immigrant cells and the earliest intrathymic progenitors express the FMS-like tyrosine kinase 3 (Flt3) receptor; however, the functional significance of this expression in the thymus is not known. We used the BM transplant model to investigate the importance of Flt3 ligand (FL) for the regeneration of the T cell compartment. We show that FL is expressed in the adult mouse thymus on the surface of perivascular fibroblasts. These cells surround the proposed thymic entry site of Flt3 receptor-positive T cell progenitors. After irradiation, perivascular FL expression is up-regulated and results in an enhanced recovery of thymic cellularity. Thymic grafting experiments confirm an intrathymic requirement for FL. Collectively, these results show that thymic stromal cell-mediated FL-Flt3 receptor interactions are important in the reconstitution of thymopoiesis early after lethal irradiation and HSCT, and provide a functional relevance to the expression of the Flt3 receptor on intrathymic T cell progenitors.


Calosota Curtis (Hymenoptera, Chalcidoidea, Eupelmidae) - review of the New World and European fauna including revision of species from the West Indies and Central and North America.
Two of three species previously classified in Calosota Curtis (Hymenoptera: Eupelmidae) from the Neotropical region are transferred to Eupelminae. Calosota eneubulus (Walker) from Galapagos Islands is transferred to Eupelmus Dalman as Eupelmus (Eupelmus) eneubulus (Walker), comb. n., and Calosota silvai (Brèthes) from Chile is transferred to Brasema Cameron as Brasema silvaicomb. n.Calosota cecidobius (Kieffer) from Argentina is retained in Calosota, with reservation, as an unrecognized species. The species of Calosota from the New World excluding South America are revised. Eleven species are recognized, including the seven newly described species Calosota albipalpussp. n. (Costa Rica, Mexico, USA, Venezuela), Calosota bicoloratasp. n. (USA), Calosota elongatasp. n. (USA), Calosota longivenasp. n. (USA), Calosota panamaensissp. n. (Panama), Calosota setosasp. n. (Bahamas, Dominican Republic, USA), and Calosota speculifronssp. n. (Costa Rica, USA). The 11 regional species and the Palaearctic species Calosota vernalis Curtis are keyed and illustrated. Calosota vernalis is not known to occur in the New World but is included in the key and diagnosed because it has been intercepted in quarantine in Canada. Calosota pseudotsugae Burks is placed in synonymy under Calosota acron (Walker), syn. n.,and Calosota kentra Burks, Calosota montana Burks and Calosota septentrionalis Hedqvist are placed in synonymy under Calosota aestivalis Curtis syn. n.Calosota modesta Bolívar y Pieltain is removed from synonymy under Calosota viridis Masi, stat. rev., and Calosota viridis, Calosota matritensis Bolívar y Pieltain, and Calosota coerulea Nikol'skaya are placed in synonymy under Calosota metallica (Gahan), syn. n.Calosota grylli Erdös is confirmed as a separate species from Calosota metallica based on features of both sexes. It is suggested that Calosota ariasi Bolívar y Pieltain may be a synonym of Calosota aestivalis, Calosota bolivari Askew may be a synonym of Calosota agrili Nikol'skaya, Calosota dusmeti Bolívar y Pieltain may be a synonym of Calosota violascens Masi, and Calosota lixobia Erdös likely is not a junior synonym of Calosota obscura Ruschka, but formal nomenclatural changes are not proposed prior to a comprehensive Palaearctic revision. Previous interpretation of the mesoscutum in Calosota and Balcha Walker is also modified to include the presence of anteroadmedian lines in addition to notauli and parapsidal lines.


Characterization of the intermediates in the reaction of mixed-valence state soluble cytochrome oxidase with oxygen at low temperatures by optical and electron-paramagnetic-resonance spectroscopy.
The reaction of soluble mixed-valence-state (a3+CuA 2+.CuB + A32+) cytochrome oxidase with O2 at low temperature was studied by optical and e.p.r. spectroscopy. The existence of three intermediates [Clore & Chance (1978) Biochem. J. 173, 799-8101] was confirmed. From the e.p.r data it is clear that cytochrome a and CuA remain in the low-spin ferric and cupric states respectively throughout the reaction. No e.p.r. signals attributable to cytochrome a3 or CuB were seen in the intermediates. The difference spectra (intermediates minus unliganded mixed-valence-state cytochrome oxidase) and absolute spectra of the three intermediates were obtained. The chemcal nature of the three intermediates is discussed in terms of their spectroscopic properties. A catalytic cycle for cytochrome oxidase is proposed.


Can an ethics officer role reduce delays in research ethics approval? A mixed-method evaluation of an improvement project.
Frustration continues to be directed at delays in gaining approvals for undertaking health research in the UK. We aimed to evaluate the impact of an ethics officer intervention on rates of favourable opinions (approval) and provisional opinions (requiring revision and resubmission) and on the time taken to reach a final opinion by research ethics committees (RECs), to characterise how the role operated in practice, and to investigate applicants' views. Mixed-method study involving (i) a 2-group, non-randomised before-and-after intervention study of RECs assigned an ethics officer and a matched comparator group; (ii) a process evaluation involving a survey of applicants and documentary analysis. 6 RECs and 3 associated ethics officers; 18 comparator RECs; REC applicants. Rates of provisional and favourable opinions between ethics officer and comparator RECs did not show a statistically significant effect of the intervention (logistic regression, p=0.26 for favourable opinions and p=0.31 for provisional opinions). Mean time to reach a decision showed a non-significant reduction (ANOVA, p=0.22) from 33.3 to 32.0 days in the ethics officer RECs compared with the comparator RECs (32.6 to 32.9 days). The survey (30% response rate) indicated applicant satisfaction and also suggested that ethics officer support might be more useful before submission. Ethics officers were successful in identifying many issues with applications, but the intervention did not function exactly as designed: in 31% of applicants, no contact between the applicants and the ethics officer took place before REC review. This study was a non-randomised comparison cohort study. Some data were missing. An ethics officer intervention, as designed and implemented in this study, did not increase the proportion of applications to RECs that were approved on first review and did not reduce the time to a committee decision.


Learning Modal-Invariant Angular Metric by Cyclic Projection Network for VIS-NIR Person Re-Identification.
Person re-identification across visible and near-infrared cameras (VIS-NIR Re-ID) has widespread applications. The challenge of this task lies in heterogeneous image matching. Existing methods attempt to learn discriminative features via complex feature extraction strategies. Nevertheless, the distributions of visible and near-infrared features are disparate caused by modal gap, which significantly affects feature metric and makes the performance of the existing models poor. To address this problem, we propose a novel approach from the perspective of metric learning. We conduct metric learning on a well-designed angular space. Geometrically, features are mapped from the original space to the hypersphere manifold, which eliminates the variations of feature norm and concentrates on the angle between the feature and the target category. Specifically, we propose a cyclic projection network (CPN) that transforms features into an angle-related space while identity information is preserved. Furthermore, we proposed three kinds of loss functions, AICAL, LAL and DAL, in angular space for angular metric learning. Multiple experiments on two existing public datasets, SYSU-MM01 and RegDB, show that performance of our method greatly exceeds the SOTA performance.


A Role for TIC55 as a Hydroxylase of Phyllobilins, the Products of Chlorophyll Breakdown during Plant Senescence.
Chlorophyll degradation is the most obvious hallmark of leaf senescence. Phyllobilins, linear tetrapyrroles that are derived from opening of the chlorin macrocycle by the Rieske-type oxygenase PHEOPHORBIDE a OXYGENASE (PAO), are the end products of chlorophyll degradation. Phyllobilins carry defined modifications at several peripheral positions within the tetrapyrrole backbone. While most of these modifications are species-specific, hydroxylation at the C32 position is commonly found in all species analyzed to date. We demonstrate that this hydroxylation occurs in senescent chloroplasts of Arabidopsis thaliana. Using bell pepper (Capsicum annuum) chromoplasts, we establish that phyllobilin hydroxylation is catalyzed by a membrane-bound, molecular oxygen-dependent, and ferredoxin-dependent activity. As these features resemble the requirements of PAO, we considered membrane-bound Rieske-type oxygenases as potential candidates. Analysis of mutants of the two Arabidopsis Rieske-type oxygenases (besides PAO) uncovered that phyllobilin hydroxylation depends on TRANSLOCON AT THE INNER CHLOROPLAST ENVELOPE55 (TIC55). Our work demonstrates a catalytic activity for TIC55, which in the past has been considered as a redox sensor of protein import into plastids. Given the wide evolutionary distribution of both PAO and TIC55, we consider that chlorophyll degradation likely coevolved with land plants.


Single-nucleus RNA-seq identifies Huntington disease astrocyte states.
Huntington Disease (HD) is an inherited movement disorder caused by expanded CAG repeats in the Huntingtin gene. We have used single nucleus RNASeq (snRNASeq) to uncover cellular phenotypes that change in the disease, investigating single cell gene expression in cingulate cortex of patients with HD and comparing the gene expression to that of patients with no neurological disease. In this study, we focused on astrocytes, although we found significant gene expression differences in neurons, oligodendrocytes, and microglia as well. In particular, the gene expression profiles of astrocytes in HD showed multiple signatures, varying in phenotype from cells that had markedly upregulated metallothionein and heat shock genes, but had not completely lost the expression of genes associated with normal protoplasmic astrocytes, to astrocytes that had substantially upregulated glial fibrillary acidic protein (GFAP) and had lost expression of many normal protoplasmic astrocyte genes as well as metallothionein genes. When compared to astrocytes in control samples, astrocyte signatures in HD also showed downregulated expression of a number of genes, including several associated with protoplasmic astrocyte function and lipid synthesis. Thus, HD astrocytes appeared in variable transcriptional phenotypes, and could be divided into several different "states", defined by patterns of gene expression. Ultimately, this study begins to fill the knowledge gap of single cell gene expression in HD and provide a more detailed understanding of the variation in changes in gene expression during astrocyte "reactions" to the disease.


Sex-specific clines support incipient speciation in a common European mammal.
Hybrid zones provide excellent opportunities to study processes and mechanisms underlying reproductive isolation and speciation. Here we investigated sex-specific clines of molecular markers in hybrid zones of morphologically cryptic yet genetically highly-diverged evolutionary lineages of the European common vole (Microtus arvalis). We analyzed the position and width of four secondary contact zones along three independent transects in the region of the Alps using maternally (mitochondrial DNA) and paternally (Y-chromosome) inherited genetic markers. Given male-biased dispersal in the common vole, a selectively neutral secondary contact would show broader paternal marker clines than maternal ones. In a selective case, for example, involving a form of Haldane's rule, Y-chromosomal clines would not be expected to be broader than maternal markers because they are transmitted by the heterogametic sex and thus gene flow would be restricted. Consistent with the selective case, paternal clines were significantly narrower or at most equal in width to maternal clines in all contact zones. In addition, analyses using maximum likelihood cline-fitting detected a shift of paternal relative to maternal clines in three of four contact zones. These patterns suggest that processes at the contact zones in the common vole are not selectively neutral, and that partial reproductive isolation is already established between these evolutionary lineages. We conclude that hybrid zone movement, sexual selection and/or genetic incompatibilities are likely associated with an unusual unidirectional manifestation of Haldane's rule in this common European mammal.


Surgical treatment of intracerebral haemorrhage.
Controversy still exists about the management of spontaneous intracerebral haemorrhage. This review summarizes our current knowledge on indications and benefits of surgery for intracerebral haemorrhage. It further describes the detailed meta-analysis of the surgical results in lobar (superficial) supratentorial intracerebral haemorrhage, summarizes the limited information on the surgical treatment of cerebellar intracerebral haemorrhage, and identifies three ongoing trials. A meta-analysis of 12 prospective randomized controlled trials of neurosurgical intervention in spontaneous intracerebral haemorrhage shows a strong trend to reduced mortality (0.85; 95% confidence interval 0.71-1.02). There is an apparent significant benefit from surgery in the three trials in which lobar intracerebral haemorrhage was identified in relation to both death and disability (0.58; 95% confidence interval 0.36 - 0.92). No surgical trial data are available for spontaneous cerebellar intracerebral haemorrhage or for any type of traumatic intracranial haemorrhage. Three trials are ongoing to resolve existing uncertainties. Clinicians are encouraged to discover if the suspected beneficial role of surgery truly exists by randomizing patients with intracerebral haemorrhage in the ongoing prospective randomized controlled trials in the following circumstances: lobar intracerebral haemorrhage (clot reaches to within 1 cm of the cortical surface), STICH II; intraventricular haemorrhage (clot maximum size of 30 ml), CLEAR IVH; deep intracerebral haemorrhage and minimal invasive surgery, MISTIE.


CircMTO1 inhibits cell proliferation and invasion by regulating Wnt/β-catenin signaling pathway in colorectal cancer.
Circular RNAs (circRNAs) play critical roles in disease incidence. However, the roles of circRNAs in colorectal cancer (CRC) progression remain largely unknown. We explored the expression of circMTO1 in CRC and elucidated the underlying molecular mechanisms. Quantitative Real-time-PCR (qRT-PCR) was used to explore circMTO1 expression in CRC tissues and cell lines. The effect of circMTO1 on the biological function of CRC cells was analyzed by Cell Counting Kit-8 (CCK-8) assay, Edu assay, colony formation assay, wound-healing assay and transwell invasion assay. Gene expression and signaling pathway were detected by qRT-PCR and Western blot. QRT-PCR showed that circMTO1 expression was significantly decreased in CRC tissues and cell lines compared with adjacent non-tumor tissues and human normal colon epithelial cell line (FHC), respectively. Patients with low circMTO1 expression were correlated with advanced TNM stage, lymph node metastasis, and poor overall survival. Function assays demonstrated that circMTO1 inhibition promoted CRC cells proliferation and invasion ability in vitro. In addition, we showed that circMTO1 inhibition could promote CRC progression via activating Wnt/β-catenin signaling pathway. We showed that circMTO1 could act as a tumor suppressor affecting the growth and invasion of CRC cells via regulating Wnt/β-catenin signaling pathway, providing a novel potential biomarker and therapeutic target for CRC treatment.


High-frequency electrical stimulation of the human skin induces heterotopical mechanical hyperalgesia, heat hyperalgesia, and enhanced responses to nonnociceptive vibrotactile input.
High-frequency electrical stimulation (HFS) of the human skin induces increased pain sensitivity in the surrounding unconditioned skin. The aim of the present study was to characterize the relative contribution of the different types of nociceptive and nonnociceptive afferents to the heterotopical hyperalgesia induced by HFS. In 17 healthy volunteers (9 men and 8 women), we applied HFS to the ventral forearm. The intensity of perception and event-related brain potentials (ERPs) elicited by vibrotactile stimuli exclusively activating nonnociceptive low-threshold mechanoreceptors and thermonociceptive stimuli exclusively activating heat-sensitive nociceptive afferents were recorded before and after HFS. The previously described mechanical hyperalgesia following HFS was confirmed by measuring the changes in the intensity of perception elicited by mechanical punctate stimuli. HFS increased the perceived intensity of both mechanical punctate and thermonociceptive stimuli applied to the surrounding unconditioned skin. The time course of the effect of HFS on the perception of mechanical and thermal nociceptive stimuli was similar. This indicates that HFS does not only induce mechanical hyperalgesia, but also induces heat hyperalgesia in the heterotopical area. Vibrotactile ERPs were also enhanced after HFS, indicating that nonnociceptive somatosensory input could contribute to the enhanced responses to mechanical pinprick stimuli. Finally, the magnitude of thermonociceptive ERPs was unaffected by HFS, indicating that type II A-fiber mechano-heat nociceptors, thought to be the primary contributor to these brain responses, do not significantly contribute to the observed heat hyperalgesia.


ITPase activity in dry blood spots is comparable with that in fresh erythrocytes.
Inosine triphosphate pyrophosphohydrolase (ITPase) catalyzing the pyrophosphohydrolysis of inosine triphosphate, deoxyinosine triphosphate and xanthosine triphosphate is involved in the metabolism and tolerance of thiopurine drugs. ITPase activity plays an important role in the prediction of toxicity to thiopurine therapy. Activities in dry blood spots were compared with fresh erythrocytes. Samples were incubated with inosine triphosphate, then inosine monophosphate was determined by a capillary electrophoresis method. Calculated enzyme activities obtained from dry blood spots were in good accordance with activity in fresh erythrocytes.


Chromosomal regions specific to pathogenic isolates of Escherichia coli have a phylogenetically clustered distribution.
We studied the ancestry of virulence-associated genes in Escherichia coli by examining chromosomal regions specific to pathogenic isolates. The four virulence determinants examined were the alpha-hemolysin (hly) loci hlyI and hlyII, the type II capsule gene cluster kps, and the P (pap) and S (sfa) fimbria gene clusters. All four loci were shown previously to be associated with pathogenicity islands of uropathogenic E. coli isolates. The hly, kps, sfa, and pap regions each have an unexpected clustered distribution among the E. coli collection of reference (ECOR) strains, but all these regions were absent from a collection of diarrheagenic E. coli isolates. Strains in the ECOR subgroup B2 typically had a combination of at least three of the four loci, and all strains in subgroup D had a copy of the kps and pap clusters. In contrast, only four strains in subgroup A had either hly, kps, sfa, or pap, and no subgroup A strains had all four together. Strains of subgroup B1 were devoid of all four virulence regions, with the exception of one isolate that had a copy of the sfa gene cluster. This phylogenetic distribution of strain-specific sequences corresponds to the ECOR groups with the largest genome size, namely, B2 and D. We propose that the pathogenicity islands are ancestral to subgroups B2 and D and were acquired after speciation, with subsequent horizontal transfer into some group A, B1, and E lineages. These results suggest that the hly, kps, sfa, and pap pathogenicity determinants may play a role in the evolution of enteric bacteria quite apart from, and perhaps with precedence over, their ability to cause disease.


A multifunctional bio-patch crosslinked with glutaraldehyde for enhanced mechanical performance, anti-coagulation properties, and anti-calcification properties.
Bio-patches for the treatment of valvular disease have been evaluated in clinical trials. It has been shown that failure of these devices, occurring within a few years of implantation, may be due to cytotoxicity, immune response, calcification and thrombosis. Some of these effects may be due to the glutaraldehyde crosslinking process used in the preparation of the materials. A number of studies have focused on strategies to control calcification, while others have concentrated on the prevention of micro-thrombus formation. In the present work, we have introduced amino-terminated poly(ethylene glycol) (NH2-PEG-NH2) as an intermolecular bridge, which not only eliminates free aldehyde groups to prevent calcification, but also introduces sites for the attachment of anticoagulant molecules. Furthermore, PEG, itself a hydrophilic polymer with good biocompatibility, may effectively prevent protein adsorption in the early stages of blood contact leading to thrombus formation. After further covalent attachment of heparin, modified bovine pericardium (BP) showed strong anti-calcification (calcium content: 39.3 ± 3.1 μg mg-1) and anti-coagulation properties (partial thromboplastin time: >300 s). The biocompatibility and mechanical properties, important for clinical use, were also improved by modification. The strategy used in this work includes new ideas and technologies for the improvement of valve products used in the clinic.


Sprouty-4 negatively regulates cell spreading by inhibiting the kinase activity of testicular protein kinase.
TESK1 (testicular protein kinase 1) is a serine/threonine kinase that phosphorylates cofilin and plays a critical role in integrin-mediated actin cytoskeletal reorganization and cell spreading. We previously showed that TESK1 interacts with Sprouty-4 (referred to as Spry4), an inhibitor of growth factor-induced Ras/MAP (mitogen-activated protein) kinase signalling, but the functional role of this interaction has remained unknown. In the present study, we show that Spry4 inhibits the kinase activity of TESK1 by binding to it through the C-terminal cysteine-rich region. Expression of Spry4 in cultured cells suppressed integrin-mediated cell spreading, and TESK1 reversed the inhibitory effect of Spry4 on cell spreading. Furthermore, Spry4 suppressed integrin- and TESK1-mediated cofilin phosphorylation during the spreading of cells on laminin. These findings suggest that Spry4 suppresses cell spreading by inhibiting the kinase activity of TESK1. Although tyrosine phosphorylation is required for the inhibitory activity of Spry4 on a Ras/MAP kinase pathway, mutation of the corresponding tyrosine residue (Tyr-75 in human Spry4) to an alanine had no apparent effect on its inhibitory actions on TESK1 activity and cell spreading, which suggests a novel cellular function of Spry to regulate the actin cytoskeleton, independent of its inhibitory activity on the Ras/MAP kinase signalling.


Modified proenzymes as artificial substrates for proteolytic enzymes: colorimetric assay of bacterial collagenase and matrix metalloproteinase activity using modified pro-urokinase.
We describe a new principle for assessment of the activity of proteolytic enzymes of all classes and show the application of this principle for the quantitative assay of bacterial collagenase and human matrix metalloproteinases (MMPs). Central to this new principle is the presence of a proenzyme that can be activated into an active enzyme by a single proteolytic event. The regular activation sequence in the proenzyme is replaced using protein engineering by an artificial sequence recognized by the proteinase to be determined. The latter can act as an activator for the newly engineered proenzyme. In the present paper a simple colorimetric assay for the determination for MMPs is described based on this principle. With the aid of protein engineering, a modified pro-urokinase has been prepared in which the activation sequence normally recognized by plasmin (Pro-Arg-Phe-Lys upward arrowIle-Ile-Gly-Gly) has been replaced by a sequence expected to be recognized and hydrolysed by many MMPs (Arg-Pro-Leu-Gly upward arrowIle-Ile-Gly-Gly). The active urokinase resulting from activation of the modified pro-urokinase by a MMP could be measured either directly, using a specific chromogenic peptide substrate for urokinase, or indirectly via urokinase-catalysed plasminogen activation. The response of the assay to equal molar quantities of active MMPs decreases in the order MMP-2>MMP-9>MMP-1>MMP-3>MMP-7. The detection limit for MMP-9 was below 15 pM, corresponding to 3. 75x10(-15) mol per assay. Using the assay, increased MMP activity was detected in synovial tissue extracts from rheumatoid arthritis patients compared with those from osteoarthritis patients, and in stomach tumour extracts as compared with normal stomach tissue extracts.


[Using hydraulic input impedance in determination of changes in the arteries of different calibre].
In anaesthetised cats, the arterial input impedance in combination with seven-element lumped-parameter model was used to estimate the resistance change in arteries of different caliber. The results show that the method gives reasonable estimations of changes in hydraulic resistance of arterial vessels of different caliber. We found that the method of vascular input impedance permits to reveal and assess quantitatively local constrictions and dilations as well as hemodynamically insignificant stenosis of conduit arteries.


A gene-targeted approach to investigate the intestinal butyrate-producing bacterial community.
Butyrate, which is produced by the human microbiome, is essential for a well-functioning colon. Bacteria that produce butyrate are phylogenetically diverse, which hinders their accurate detection based on conventional phylogenetic markers. As a result, reliable information on this important bacterial group is often lacking in microbiome research. In this study we describe a gene-targeted approach for 454 pyrotag sequencing and quantitative polymerase chain reaction for the final genes in the two primary bacterial butyrate synthesis pathways, butyryl-CoA:acetate CoA-transferase (but) and butyrate kinase (buk). We monitored the establishment and early succession of butyrate-producing communities in four patients with ulcerative colitis who underwent a colectomy with ileal pouch anal anastomosis and compared it with three control samples from healthy colons. All patients established an abundant butyrate-producing community (approximately 5% to 26% of the total community) in the pouch within the 2-month study, but patterns were distinctive among individuals. Only one patient harbored a community profile similar to the healthy controls, in which there was a predominance of but genes that are similar to reference genes from Acidaminococcus sp., Eubacterium sp., Faecalibacterium prausnitzii and Roseburia sp., and an almost complete absence of buk genes. Two patients were greatly enriched in buk genes similar to those of Clostridium butyricum and C. perfringens, whereas a fourth patient displayed abundant communities containing both genes. Most butyrate producers identified in previous studies were detected and the general patterns of taxa found were supported by 16S rRNA gene pyrotag analysis, but the gene-targeted approach provided more detail about the potential butyrate-producing members of the community. The presented approach provides quantitative and genotypic insights into butyrate-producing communities and facilitates a more specific functional characterization of the intestinal microbiome. Furthermore, our analysis refines but and buk reference annotations found in central databases.


Correlation between the expression of DNMT1, and GSTP1 and APC, and the methylation status of GSTP1 and APC in association with their clinical significance in prostate cancer.
The aim of the present study was to investigate the correlation between the expression of DNA (cytosine‑5)‑methyltransferase 1 (DNMT1), glutathione S‑transferase‑P1 (GSTP1) and adenomatous polyposis coli (APC), and the methylation status of GSTP1 and APC in prostate cancer (PCa) and benign prostatic hyperplasia (BPH), and to examine its clinical significance. Immunohistochemistry and reverse transcription‑polymerase chain reaction (RT‑PCR) was used to detect the expression of DNMT1, GSTP1 and APC in 56 samples of PCa tissue and 10 samples of BPH tissue. Methylation‑specific‑PCR was used to detect the methylation status of the CpG island promoters of GSTP1 and APC. The positive rate of expression of DNMT1 in poorly‑differentiated PCa, moderately‑differentiated PCa, well‑differentiated PCa and BPH was 86.7%, 70.6%, 55.6% and 30.0%, respectively (P<0.05); for GSTP1, the positive rate was 13.3%, 29.4%, 44.4% and 90.0%, respectively (P<0.05); and for APC, the positive rate was 23.3%, 47.6%, 55.6% and 70.0%, respectively (P<0.05). The correlation coefficient for the association between the expression of DNMT1 and GSTP1 was ‑0.891 (P<0.05). Between the expression of DNMT1 and APC, the correlation coefficient was ‑0.721 (P<0.05). GSTP1 and APC were hypermethylated in the majority of PCa tissue samples. The positive rate of methylation of these genes in poorly‑differentiated PCa was 83.3% and 73.3%, respectively. By contrast, hypomethylation (or demethylation) was observed in BPH samples, in which the positive rate of methylation was 10.0% and 20.0%, respectively (P<0.05). The increased expression of DNMT1, and the reduced expression of GSTP1 and APC have an important role in the development of PCa. Due to the high expression of DNMT1 mRNA, it is likely that the hypermethylation of CpG islands contributed to the silencing of GSTP1 and APC in PCa tissues.


Brief interpersonal psychotherapy for depressed mothers whose children are receiving psychiatric treatment.
Depressed mothers of children with psychiatric illness struggle with both their own psychiatric disorder and the demands of caring for ill children. When maternal depression remains untreated, mothers suffer, and psychiatric illness in their offspring is less likely to improve. This randomized, controlled trial compared the interpersonal psychotherapy for depressed mothers (IPT-MOMS), a nine-session intervention based on standard interpersonal psychotherapy, to treatment as usual for depressed mothers with psychiatrically ill offspring. Forty-seven mothers meeting DSM-IV criteria for major depression were recruited from a pediatric mental health clinic where their school-age children were receiving psychiatric treatment and randomly assigned to IPT-MOMS (N=26) or treatment as usual (N=21). Mother-child pairs were assessed at three time points: baseline, 3-month follow-up, and 9-month follow-up. Child treatment was not determined by the study. Compared to subjects assigned to treatment as usual, subjects assigned to IPT-MOMS showed significantly lower levels of depression symptoms, as measured by the Hamilton Depression Rating Scale, and higher levels of functioning, as measured by the Global Assessment of Functioning, at 3-month and 9-month follow-ups. Compared to the offspring of mothers receiving treatment as usual, the offspring of mothers assigned to IPT-MOMS showed significantly lower levels of depression as measured by the Children's Depressive Inventory at the 9-month follow-up. Assignment to IPT-MOMS was associated with reduced levels of maternal symptoms and improved functioning at the 3- and 9-month follow-ups compared to treatment as usual. Maternal improvement preceded improvement in offspring, suggesting that maternal changes may mediate child outcomes.


Primary sensory and motor cortex excitability are co-modulated in response to peripheral electrical nerve stimulation.
Peripheral electrical stimulation (PES) is a common clinical technique known to induce changes in corticomotor excitability; PES applied to induce a tetanic motor contraction increases, and PES at sub-motor threshold (sensory) intensities decreases, corticomotor excitability. Understanding of the mechanisms underlying these opposite changes in corticomotor excitability remains elusive. Modulation of primary sensory cortex (S1) excitability could underlie altered corticomotor excitability with PES. Here we examined whether changes in primary sensory (S1) and motor (M1) cortex excitability follow the same time-course when PES is applied using identical stimulus parameters. Corticomotor excitability was measured using transcranial magnetic stimulation (TMS) and sensory cortex excitability using somatosensory evoked potentials (SEPs) before and after 30 min of PES to right abductor pollicis brevis (APB). Two PES paradigms were tested in separate sessions; PES sufficient to induce a tetanic motor contraction (30-50 Hz; strong motor intensity) and PES at sub motor-threshold intensity (100 Hz). PES applied to induce strong activation of APB increased the size of the N(20)-P(25) component, thought to reflect sensory processing at cortical level, and increased corticomotor excitability. PES at sensory intensity decreased the size of the P25-N33 component and reduced corticomotor excitability. A positive correlation was observed between the changes in amplitude of the cortical SEP components and corticomotor excitability following sensory and motor PES. Sensory PES also increased the sub-cortical P(14)-N(20) SEP component. These findings provide evidence that PES results in co-modulation of S1 and M1 excitability, possibly due to cortico-cortical projections between S1 and M1. This mechanism may underpin changes in corticomotor excitability in response to afferent input generated by PES.


Attributable causes of breast cancer and ovarian cancer in china: reproductive factors, oral contraceptives and hormone replacement therapy.
To provide an evidence-based, consistent assessment of the burden of breast cancer attributable to reproductive factors (RFs, including nulliparity, mean number of children, age at first birth and breastfeeding), use of oral contraceptives (OCs, restricted to the age group of 15-49 years), and hormone replacement therapy (HRT), as well as of the burden of ovarian cancer attributable to the mean number of children in China in 2005. We derived the prevalence of these risk factors and the relative risk of breast and ovarian cancer from national surveys or large-scale studies conducted in China. In the case of RFs, we compared the exposure distributions in 2001 and counterfactual exposure. Exposure of RFs in 2001 was found to account for 6.74% of breast cancer, corresponding to 9,617 cases and 2,769 deaths, and for 2.78% of ovarian cancer (711 cases, 294 deaths). The decrease in mean number of children alone was responsible for 1.47% of breast cancer and 2.78% of ovarian cancer. The prevalence of OC use was 1.74% and the population attributable fraction (PAF) of breast cancer was 0.71%, corresponding to 310 cases and 90 deaths. The PAF of breast cancer due to HRT was 0.31%, resulting in 297 cases and 85 deaths. RFs changes in China contributed to a sizable fraction of breast and ovarian cancer incidence and mortality, whereas HRT and OCs accounted for relatively low incidence of breast cancer in China.


Methodological Challenges in Randomized Controlled Trials on Smartphone-Based Treatment in Psychiatry: Systematic Review.
Smartphone-based technology is developing at high speed, and many apps offer potential new ways of monitoring and treating a range of psychiatric disorders and symptoms. However, the effects of most available apps have not been scientifically investigated. Within medicine, randomized controlled trials (RCTs) are the standard method for providing the evidence of effects. However, their rigidity and long time frame may contrast with the field of information technology research. Therefore, a systematic review of methodological challenges in designing and conducting RCTs within mobile health is needed. This systematic review aimed to (1) identify and describe RCTs investigating the effect of smartphone-based treatment in adult patients with a psychiatric diagnosis, (2) discuss methodological challenges in designing and conducting individual trials, and (3) suggest recommendations for future trials. A systematic search in English was conducted in PubMed, PsycINFO, and EMBASE up to August 12, 2019. The search terms were (1) psychiatric disorders in broad term and for specific disorders AND (2) smartphone or app AND (3) RCT. The Consolidated Standards of Reporting Trials electronic health guidelines were used as a template for data extraction. The focus was on trial design, method, and reporting. Only trials having sufficient information on diagnosis and acceptable diagnostic procedures, having a smartphone as a central part of treatment, and using an RCT design were included. A total of 27 trials comprising 3312 patients within a range of psychiatric diagnoses were included. Among them, 2 trials were concerning drug or alcohol abuse, 3 psychosis, 10 affective disorders, 9 anxiety and posttraumatic stress disorder, 1 eating disorder, and 1 attention-deficit/hyperactivity disorder. In addition, 1 trial used a cross-diagnostic design, 7 trials included patients with a clinical diagnosis that was subsequently assessed and validated by the researchers, and 11 trials had a sample size above 100. Generally, large between-trial heterogeneity and multiple approaches to patient recruitment, diagnostic procedures, trial design, comparator, outcome measures, and analyses were identified. Only 5 trials published a trial protocol. Furthermore, 1 trial provided information regarding technological updates, and only 18 trials reported on the conflicts of interest. No trial addressed the ethical aspects of using smartphones in treatment. This first systematic review of the methodological challenges in designing and conducting RCTs investigating smartphone-based treatment in psychiatric patients suggests an increasing number of trials but with a lower quality compared with classic medical RCTs. Heterogeneity and methodological issues in individual trials limit the evidence. Methodological recommendations are presented.


Genes to remember.
It has been known for several decades that the formation of long-term memory requires gene expression. In recent years, the use of genetic and molecular approaches has led to the identification and characterization of genes and molecules that play a fundamental role in the biological mechanisms underlying learning and memory. From these studies, it appears that molecules and molecular mechanisms essential for the process of memory have been conserved throughout evolution. The cyclic AMP (cAMP)-dependent activation pathway and a cAMP-dependent cascade of gene expression have been shown to be essential for memory formation in Aplysia californica, Drosophila melanogaster and rodents. Moreover, members of the transcription factor family cAMP response element binding proteins (CREBs) seem to represent key molecules for transforming incoming information into long-term memory. Here, we review the studies showing that conserved molecules and biological mechanisms are engaged in simple and complex forms of memory.


Lentiviral vectors: progress and potential.
Lentiviral vectors are continuing to generate great interest as gene vectors for possible gene therapy in vivo. Over the past year, considerable progress has been made in demonstrating gene delivery to a wide variety of tissues, taking advantage of their unique capability of integrating the desired gene into the chromosomes of the target cell. The basic science of lentivirus vector production has expanded and their relative ease of utility is reflected by the broadening range of laboratories reporting successful gene transfer using them. Important new findings on RNA processing within the cell have come from studies of vectors which have direct relevance to the molecular biology of HIV itself. Although none have been used in clinical trials in vivo, it can only be a matter of time before this occurs. This article reviews publications over the last 12 months, highlighting the major achievements in this field.


An Australian study on feigned mTBI using the Inventory of Problems - 29 (IOP-29), its Memory Module (IOP-M), and the Rey Fifteen Item Test (FIT).
We investigated the classification accuracy of the Inventory of Problems - 29 (IOP-29), its newly developed memory module (IOP-M) and the Fifteen Item Test (FIT) in an Australian community sample (N = 275). One third of the participants (n = 93) were asked to respond honestly, two thirds were instructed to feign mild TBI. Half of the feigners (n = 90) were coached to avoid detection by not exaggerating, half were not (n = 92). All measures successfully discriminated between honest responders and feigners, with large effect sizes (d ≥ 1.96). The effect size for the IOP-29 (d ≥ 4.90), however, was about two-to-three times larger than those produced by the IOP-M and FIT. Also noteworthy, the IOP-29 and IOP-M showed excellent sensitivity (>90% the former, > 80% the latter), in both the coached and uncoached feigning conditions, at perfect specificity. Instead, the sensitivity of the FIT was 71.7% within the uncoached simulator group and 53.3% within the coached simulator group, at a nearly perfect specificity of 98.9%. These findings suggest that the validity of the IOP-29 and IOP-M should generalize to Australian examinees and that the IOP-29 and IOP-M likely outperform the FIT in the detection of feigned mTBI.


Serine/threonine kinase 17A is a novel candidate for therapeutic targeting in glioblastoma.
STK17A is a relatively uncharacterized member of the death-associated protein family of serine/threonine kinases which have previously been associated with cell death and apoptosis. Our prior work established that STK17A is a novel p53 target gene that is induced by a variety of DNA damaging agents in a p53-dependent manner. In this study we have uncovered an additional, unanticipated role for STK17A as a candidate promoter of cell proliferation and survival in glioblastoma (GBM). Unexpectedly, it was found that STK17A is highly overexpressed in a grade-dependent manner in gliomas compared to normal brain and other cancer cell types with the highest level of expression in GBM. Knockdown of STK17A in GBM cells results in a dramatic alteration in cell shape that is associated with decreased proliferation, clonogenicity, migration, invasion and anchorage independent colony formation. STK17A knockdown also sensitizes GBM cells to genotoxic stress. STK17A overexpression is associated with a significant survival disadvantage among patients with glioma which is independent of age, molecular phenotype, IDH1 mutation, PTEN loss, and alterations in the p53 pathway and partially independent of grade. In summary, we demonstrate that STK17A provides a proliferative and survival advantage to GBM cells and is a potential target to be exploited therapeutically in patients with glioma.


[Barriers to implementation of cervical cancer screening: a household survey in the coverage area of the Family Health Program in Nova Iguaçu, Rio de Janeiro State, Brazil].
The aim of this study was to analyze barriers to access to cervical cancer screening under the Family Health Program in Nova Iguacu, Rio de Janeiro State, Brazil. Through a household survey, we interviewed 281 women 20 to 59 years of age. To assess barriers to the Pap smear, a Portuguese version of the Champion's Health Belief Model Scale was used. Fear related to the test results (39.85%, 95%CI: 34.09-45.61) and the professional examiner (31.31%, 95%CI: 25.86-36.77), shame (39.85%, 95%CI: 34.09-45.61), and forgetting to schedule the test (32.02%, 95%CI: 26.53-37.51) were reported as the main barriers to access. Since the constraints varied according to the population's socio-demographic characteristics, organization of cervical cancer screening should be tailored accordingly.


Association of Race and Ethnicity With Comorbidities and Survival Among Patients With COVID-19 at an Urban Medical Center in New York.
As of May 11, 2020, there have been more than 290 000 deaths worldwide from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19). Risk-adjusted differences in outcomes among patients of differing ethnicity and race categories are not well characterized. To investigate whether presenting comorbidities in patients with COVID-19 in New York City differed by race/ethnicity and whether case fatality rates varied among different ethnic and racial groups, controlling for presenting comorbidities and other risk factors. This cohort study included 5902 patients who presented for care to the Montefiore Medical Center, a large urban academic medical center in the Bronx, New York, between March 14 and April 15, 2020, and tested positive for SARS-CoV-2 on reverse transcription quantitative polymerase chain reaction assay. Final data collection was April 27, 2020. Patient characteristics, including self-identified ethnicity/race, age, sex, socioeconomic status, and medical comorbidities, were tabulated. Overall survival. Associations between patient demographic characteristics, comorbidities, and race/ethnicity were examined using χ2 tests, and the association with survival was assessed using univariable and multivariable Cox proportional hazards regression, based on time from positive COVID-19 test. Of 9268 patients who were tested, 5902 ethnically diverse patients (63.7%) had SARS-CoV-2. Of these, 3129 patients (53.0%) were women, and the median (interquartile range) age was 58 (44-71) years. A total of 918 patients (15.5%) died within the study time frame. Overall, 1905 patients (32.3%) identified as Hispanic; 1935 (32.8%), non-Hispanic Black; 509 (8.6%), non-Hispanic White; and 171 (2.9%), Asian; the death rates were 16.2% (309), 17.2% (333), 20.0% (102), and 17.0% (29), respectively (P = .25). Hispanic and non-Hispanic Black patients had a higher proportion of more than 2 medical comorbidities with 654 (34.3%) and 764 (39.5%), respectively, compared with 147 (28.9%) among non-Hispanic White patients (P < .001). Hispanic and non-Hispanic Black patients were also more likely to test positive for COVID-19 than White patients, with 1905 of 2919 Hispanic patients (65.3%), 1935 of 2823 non-Hispanic Black patients (68.5%), and 509 of 960 non-Hispanic White patients (53.0%) having positive test results for SARS-CoV-2 (P < .001). While controlling for age, sex, socioeconomic status and comorbidities, patients identifying as Hispanic (hazard ratio, 0.77; 95% CI, 0.61-0.98; P = .03) or non-Hispanic Black (hazard ratio, 0.69; 95% CI, 0.55-0.87; P = .002) had slightly improved survival compared with non-Hispanic White patients. In this cohort study of patients with COVID-19 who presented for care at the same urban medical center, non-Hispanic Black and Hispanic patients did not experience worse risk-adjusted outcomes compared with their White counterparts. This finding is important for understanding the observed population differences in mortality by race/ethnicity reported elsewhere.


Genetic associations with micronutrient levels identified in immune and gastrointestinal networks.
The discovery of vitamins and clarification of their role in preventing frank essential nutrient deficiencies occurred in the early 1900s. Much vitamin research has understandably focused on public health and the effects of single nutrients to alleviate acute conditions. The physiological processes for maintaining health, however, are complex systems that depend upon interactions between multiple nutrients, environmental factors, and genetic makeup. To analyze the relationship between these factors and nutritional health, data were obtained from an observational, community-based participatory research program of children and teens (age 6-14) enrolled in a summer day camp in the Delta region of Arkansas. Assessments of erythrocyte S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), plasma homocysteine (Hcy) and 6 organic micronutrients (retinol, 25-hydroxy vitamin D3, pyridoxal, thiamin, riboflavin, and vitamin E), and 1,129 plasma proteins were performed at 3 time points in each of 2 years. Genetic makeup was analyzed with 1 M SNP genotyping arrays, and nutrient status was assessed with 24-h dietary intake questionnaires. A pattern of metabolites (met_PC1) that included the ratio of erythrocyte SAM/SAH, Hcy, and 5 vitamins were identified by principal component analysis. Met_PC1 levels were significantly associated with (1) single-nucleotide polymorphisms, (2) levels of plasma proteins, and (3) multilocus genotypes coding for gastrointestinal and immune functions, as identified in a global network of metabolic/protein-protein interactions. Subsequent mining of data from curated pathway, network, and genome-wide association studies identified genetic and functional relationships that may be explained by gene-nutrient interactions. The systems nutrition strategy described here has thus associated a multivariate metabolite pattern in blood with genes involved in immune and gastrointestinal functions.


Ex vivo expansion of CD8+CD56+ and CD8+CD56- natural killer T cells specific for MUC1 mucin.
Prostate cancers express MUC1, but nearly all metastatic cells lack HLA class I molecules. Thus, a lymphocyte population that can sense its antigenic environment, while also able to react to stimuli of natural killer (NK) cells, may be a more versatile effector cell population for antitumor immune responses. Herein, we report that tumor-specific MUC1 peptide, interleukin 2, and interleukin 12 act synergistically to stimulate the ex vivo expansion of CD8(+)CD56(-) T cells and CD8(+)CD56(+) natural killer T (NKT) cells from the peripheral blood mononuclear cells of prostate cancer patients, as well as healthy male and female donors. Both the CD56(+) NKT cells and CD56(-) T cells lysed allogeneic mucin-bearing target cells, as well as NK target cells, but not lymphokine-activated killer target cells. However, the CD56(+) NKT cells displayed a 2-fold greater cytolytic activity than the CD56(-) T cells. The mucin-specific cytolytic activity and NK cytolytic activities for both lymphocyte populations were independent of HLA class I and CD1 molecules. The CD56(-) T cells up-regulated CD56 with continued antigenic stimulation in the presence of interleukin 12, suggesting that CD8(+)CD56(-) T cells are NKT cells. However, CD56(+) NKT cells expand poorly to continued stimulation. All mucin-stimulated NKT cells exhibited the activated/memory CD45RO phenotype. The NKT cell lines express the alpha/beta T-cell receptor (TCR). The TCR repertoire was limited and varied with cell line, but was not the V alpha 24V beta 11 TCR typically associated with NKT cells. Whereas CD161 is generally considered a marker of NKT cells, the mucin-stimulated NKT cells did not express this marker. Thus, we have described two phenotypically distinct NKT types that do not display a biased TCR repertoire, but do display specificity for a tumor-specific peptide antigen (CTL-like activity), as well as HLA class I-deficient target cells (NK-like activity).


Anti-oxLDL antibody isotype levels, as potential markers for progressive atherosclerosis in APOE and APOECD40L mice.
In humans and animal models of atherosclerosis, antibodies against oxidized LDL have been associated with atherosclerotic lesion development. It has been suggested that IgM anti-oxLDL antibodies are anti-atherogenic, whereas IgG anti-oxLDL antibodies are pro-atherogenic. In this study, we examined the relation between IgM and IgG antibody levels and atherosclerosis severity in APOE(-/-)CD40L(-/-) mice, which are deficient for IgG and develop moderate advanced atherosclerosis, and compared results with mice developing severe (APOE(-/-)) or no atherosclerosis (C57Bl/6). Mice were followed in time for anti-oxLDL antibodies while on high-fat diet or normal chow. Anti-oxLDL antibody levels were determined by ELISA. Results revealed that 24-week-old APOE(-/-)CD40L(-/-) mice had enhanced IgM anti-oxLDL antibody levels when compared with wild-type mice, but similar levels to those of APOE(-/-) mice. As expected, IgG anti-oxLDL antibody levels were almost absent in APOE(-/-)CD40L(-/-) mice. The transition from early to advanced lesions in APOE(-/-) mice was reflected by elevated IgM anti-oxLDL antibody levels. IgM anti-oxLDL levels did not further increase during progression to more advanced lesions. No relation was found between IgG anti-oxLDL levels and atherosclerosis severity. In conclusion, the severity of advanced atherosclerosis in mice is not reflected by IgM and/or IgG anti-oxLDL antibody levels. Furthermore, less advanced atherosclerotic lesion development in APOE(-/-)CD40L(-/-) mice does not seem to be the result of higher levels of protective IgM anti-oxLDL antibodies. Therefore, our study does not support the idea that the previously observed inconsistency in the relation between anti-oxLDL and atherosclerosis severity is due to differences in antibody isotypes.


LncRNA MIR31HG promotes cell proliferation and invasive properties of the MCF-7 cell line by regulation of receptor-interacting serine-threonine kinase 4.
LncRNA MIR31HG is involved in many types of cancers, while its roles in breast cancer are still unknown. The current study aimed to explore the function of lncRNA MIR31HG in breast cancer and the underlying mechanisms. Stable expression cell lines were constructed by using lentivirus particles. MTT assay was used to determine cell viability. Wound healing and Transwell assay were used to determine cell migration and invasion, respectively. The changes in biomarkers were determined by using qPR-PCT and Western blotting, respectively. BALB/c nude mice were used to generate a xenograft mouse model. MIR31HG regulated cell proliferation, migration and invasion in MCF7 cells. Besides, MIR31HG regulated N-Cadherin, Vimentin, and E-Cadherin. MIR31HG positively regulated receptor-interacting serine-threonine kinase 4 (RIPK4), as supported by the fact that knockdown of MIR31HG suppressed RIPK4, and the knockdown of RIPK4 did not affect MIR31HG. Additionally, we found that RIPK4 regulated cell proliferation, migration and invasion in MCF7 cells. The changes in RIPK4 regulated N-Cadherin, Vimentin, and E-Cadherin. Consistently, in vivo studies showed that the knockdown of MIR31HG or RIPK4 reduced tumor size in xenograft animal models. The roles of lncRNA MIR31HG in breast cancer were associated with its regulatory effects against RIPK4.


Perceptions and experiences of nurses in Turkey about verbal abuse in clinical settings.
To determine the frequency and sources of verbal abuse against nurses working in clinical settings in different hospitals in the last 12 months, to identify nurses' perceptions about verbal abuse, and to determine types of emotions experienced by nurses who had encountered verbal abuse. This descriptive study was conducted in May 2001 in Turkey. Data were collected from 467 nurses working in various clinical settings in three hospitals. The instrument was a 23-item questionnaire on verbal abuse. Data were evaluated using frequency and descriptive statistics. The findings revealed that the majority of nurses had experienced verbal abuse in the last 12 months (86.7%, n = 405). Most nurses (92%) reported that verbal abuse negatively affected their morale. The most common sources of verbal abuse were patients' relatives and patients themselves. Workplace verbal abuse events cannot always be anticipated. Contingency plans to handle potential situations should be developed.


In Situ Activatable Nitrobenzene-Cysteine-Copper(II) Nano-complexes for Programmed Photodynamic Cancer Therapy.
Photodynamic therapy (PDT) has been used to reduce cancerous and precancerous cells via reactive oxygen species (ROS) generation from photosensitizers. Numerous photosensitizers are available today to treat a variety of diseases, but their therapeutic efficacy is hindered within the tumor microenvironment, and there are safety concerns associated with their non-specific activation. In this work, we disclosed a nano-therapeutic based on in situ activatable nitrobenzene-cysteine-copper(II) nano-complexes (NCCNs) that work within cancer cells. Among the NCCNs, CyP shows outstanding potential as a promising candidate for programmed photodynamic cancer therapy with its unique properties such as (i) bright near-infrared imaging, (ii) chemodynamic therapeutic effect, (iii) photodynamic therapeutic effect (types I and II), and (iv) anti-cancer effect by anti-angiogenesis in early cancer stage under light. Overall, this work opens up exciting possibilities for the development of innovative and effective treatments for cancer, paving the way for future advancements in the clinical medicine field.


Adherence of pregnant women to Nordic dietary guidelines in relation to postpartum weight retention: results from the Norwegian Mother and Child Cohort Study.
Pregnancy is a major life event for women and often connected with changes in diet and lifestyle and natural gestational weight gain. However, excessive weight gain during pregnancy may lead to postpartum weight retention and add to the burden of increasing obesity prevalence. Therefore, it is of interest to examine whether adherence to nutrient recommendations or food-based guidelines is associated with postpartum weight retention 6 months after birth. This analysis is based on data from the Norwegian Mother and Child Cohort Study (MoBa) conducted by the Norwegian Institute of Public Health. Diet during the first 4-5 months of pregnancy was assessed by a food-frequency questionnaire and maternal weight before pregnancy as well as in the postpartum period was assessed by questionnaires. Two Healthy Eating Index (HEI) scores were applied to measure compliance with either the official Norwegian food-based guidelines (HEI-NFG) or the Nordic Nutrition Recommendations (HEI-NNR) during pregnancy. The considered outcome, i.e. weight retention 6 months after birth, was modelled in two ways: continuously (in kg) and categorically (risk of substantial postpartum weight retention, i.e. ≥ 5% gain to pre-pregnancy weight). Associations between the HEI-NFG and HEI-NNR score with postpartum weight retention on the continuous scale were estimated by linear regression models. Relationships of both HEI scores with the categorical outcome variable were evaluated using logistic regression. In the continuous model without adjustment for gestational weight gain (GWG), the HEI-NFG score but not the HEI-NNR score was inversely related to postpartum weight retention. However, after additional adjustment for GWG as potential intermediate the HEI-NFG score was marginally inversely and the HEI-NNR score was inversely associated with postpartum weight retention. In the categorical model, both HEI scores were inversely related with risk of substantial postpartum weight retention, independent of adjustment for GWG. Higher adherence to either the official Norwegian food guidelines or possibly also to Nordic Nutrition Recommendations during pregnancy appears to be associated with lower postpartum weight retention.