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Background: Probiotics and zinc are commonly used and beneficial in pig production. This work aimed to assess the effect of probiotic and zinc on brush-border enzyme activity and histoenzymatic study of the small intestine in pre and post-weaned piglets. Methods: Eighteen LWY piglets were divided equally into control and treatment groups. The piglets were maintained in standard management conditions and were weaned at 28 days of age. The treatment group of piglets fed a mixture of probiotics orally @ 1.25 × 109 CFU/day and zinc @ 2000 ppm/day from birth to 10 days of age. At three different age-groups viz. day 20 (pre-weaning), day 30 (weaning) and day 60 (post-weaning), the animals were sacrificed. For disaccharidase enzyme estimation, the mucosal brush border of the small intestine was scrapped off and the experiment was conducted. For histoenzymatic assay, the small intestine samples were preserved in liquid nitrogen at -196C immediately after sacrifice. They were sectioned at 10μm thickness maintained at -20C and stained for different histochemical staining. The statistical analysis of the data using the appropriate statistical tests was also conducted. Result: The activity of different brush-border enzymes such as maltase, sucrase and lactase was more in the treatment group of piglets. The activity of different histochemical enzymes such as alkaline phosphatase, acid phosphatase, adenosine tri-phosphatase and non-specific esterase was increased in the treated group of piglets. [ABSTRACT FROM AUTHOR]

The present study investigates the self healing behavior of both pristine and defected single layer graphene using a molecular dynamic simulation. Single layer graphene containing various defects such as preexisting vacancies and differently oriented pre-existing cracks were subjected to uniaxial tensile loading till fracture occurred. Once the load was relaxed, the graphene was found to undergo self healing. It was observed that this self healing behaviour of cracks holds irrespective of the nature of pre-existing defects in the graphene sheet. Cracks of any length were found to heal provided the critical crack opening distance lies within 0.3-0.5 nm for a pristine sheet and also for a sheet with pre-existing defects. Detailed bond length analysis of the graphene sheet was done to understand the mechanism of self healing of graphene. The paper also discusses the immense potential of the self healing phenomena of graphene in the field of graphene based sub-nano sensors for crack sensing.

Background: Socioecological factors are associated with key health behaviors that are critical for weight management, and major life events may disrupt engagement in these behaviors. However, the influence of socioecological factors on health behaviors in the midst of major life events is not clear and is difficult to study due to the random and sporadic nature of their occurrence. The COVID-19 pandemic provided a unique opportunity to study a major life event and its impacts on diet, physical activity, and body weight., Objective: This cross-sectional study aimed to investigate associations between socioecological factors (environmental, interpersonal, and individual) and self-reported weight change during a major life event using data collected during the COVID-19 pandemic, and whether the associations were mediated through self-reported changes in eating and physical activity behaviors., Methods: Participants self-reported socioecological factors, weight change, and changes in eating behaviors (EB) and physical activity (PA) via online questionnaires between December 2020 and October 2021. Changes in EB and PA were measured using scales with higher scores reflecting more positive changes during the COVID-19 pandemic., Results: Participants ( n  = 1283) were mostly female (84.9%) with age 52.1 ± 14.1 years (mean ± SD) and BMI of 32.9 ± 8.2 kg/m 2 . Stronger healthy eater and exercise identities (individual factors) were associated with higher EB scores (EBS) and PA scores (PAS), respectively ( p 's < 0.00001). Less discouragement for healthy eating by family/friends (interpersonal factor) was associated with higher EBS ( p  = 0.002). Higher EBS and PAS were associated with weight loss. The indirect effect of healthy eater identity (-0.72; 95% CI: -0.90, -0.55) and discouragement for diet (0.07; 95% CI: 0.03, 0.12) on weight change through EBS were significant, as was the indirect effect of exercise identity (-0.25; 95% CI: -0.35, -0.15) on weight change through PAS., Conclusions: Stronger identities and less discouragement from family/friends may support health promoting behaviors and weight loss during a major life event, as well as identify additional behavioral targets for lifestyle interventions., Clinical Trial Registration: IWCR was registered at ClinicalTrials.gov (NCT04907396)., Competing Interests: Dr. Roberts founded the iDiet, a web‐based behavioral weight loss program (www.theidiet.com) and is a Board member of Danone. Dr. Peters received grants/contracts from the American Pecan Council and McCormick Science Institute, and also received consulting fees from the University of Alabama Birmingham. The remaining authors have no relevant conflicts of interest to disclose., (© 2024 The Author(s). Obesity Science & Practice published by World Obesity and The Obesity Society and John Wiley & Sons Ltd.)

The major gram-positive pathogen group A Streptococcus (GAS) is a model organism for studying microbial epidemics as it causes waves of infections. Since 1980, several GAS epidemics have been ascribed to the emergence of clones producing increased amounts of key virulence factors such as streptolysin O (SLO). Herein, we sought to identify mechanisms underlying our recently identified temporal clonal emergence among emm4 GAS, given that emergent strains did not produce augmented levels of virulence factors relative to historic isolates. By creating and analyzing isoallelic strains, we determined that a conserved mutation in a previously undescribed gene encoding a putative carbonic anhydrase was responsible for the defective in vitro growth observed in the emergent strains. We also identified that the emergent strains survived better inside macrophages and killed macrophages at lower rates than the historic strains. Via the creation of isogenic mutant strains, we linked the emergent strain "survival" phenotype to the downregulation of the SLO encoding gene and upregulation of the msrAB operon which encodes proteins involved in defense against extracellular oxidative stress. Our findings are in accord with recent surveillance studies which found a high ratio of mucosal (i.e., pharyngeal) relative to invasive infections among emm4 GAS. Since ever-increasing virulence is unlikely to be evolutionarily advantageous for a microbial pathogen, our data further understanding of the well-described oscillating patterns of virulent GAS infections by demonstrating mechanisms by which emergent strains adapt a "survival" strategy to outcompete previously circulating isolates., Competing Interests: The authors declare no conflict of interest.

Group A streptococcal (GAS) strains causing severe, invasive infections often have mutations in the control of virulence two-component regulatory system (CovRS) which represses capsule production, and high-level capsule production is considered critical to the GAS hypervirulent phenotype. Additionally, based on studies in emm1 GAS, hyperencapsulation is thought to limit transmission of CovRS-mutated strains by reducing GAS adherence to mucosal surfaces. It has recently been identified that about 30% of invasive GAS strains lacks capsule, but there are limited data regarding the impact of CovS inactivation in such acapsular strains. Using publicly available complete genomes ( n = 2,455) of invasive GAS strains, we identified similar rates of CovRS inactivation and limited evidence for transmission of CovRS-mutated isolates for both encapsulated and acapsular emm types. Relative to encapsulated GAS, CovS transcriptomes of the prevalent acapsular emm types emm28 , emm87 , and emm89 revealed unique impacts such as increased transcript levels of genes in the emm /mga region along with decreased transcript levels of pilus operon-encoding genes and the streptokinase-encoding gene ska . CovS inactivation in emm87 and emm89 strains, but not emm28 , increased GAS survival in human blood. Moreover, CovS inactivation in acapsular GAS reduced adherence to host epithelial cells. These data suggest that the hypervirulence induced by CovS inactivation in acapsular GAS follows distinct pathways from the better studied encapsulated strains and that factors other than hyperencapsulation may account for the lack of transmission of CovRS-mutated strains. IMPORTANCE Devastating infections due to group A streptococci (GAS) tend to occur sporadically and are often caused by strains that contain mutations in the control of virulence regulatory system (CovRS). In well-studied emm1 GAS, the increased production of capsule induced by CovRS mutation is considered key to both hypervirulence and limited transmissibility by interfering with proteins that mediate attachment to eukaryotic cells. Herein, we show that the rates of covRS mutations and genetic clustering of CovRS-mutated isolates are independent of capsule status. Moreover, we found that CovS inactivation in multiple acapsular GAS emm types results in dramatically altered transcript levels of a diverse array of cell-surface protein-encoding genes and a unique transcriptome relative to encapsulated GAS. These data provide new insights into how a major human pathogen achieves hypervirulence and indicate that factors other than hyperencapsulation likely account for the sporadic nature of the severe GAS disease., Competing Interests: The authors declare no conflict of interest.

Introduction: the management of an acute spinal cord injury remains controversial. The patient of acute spinal cord injury undergoes several phases of care beginning with the initial trauma management, surgical intervention, and perioperative medical management. The aim of this study was to evaluate the neurological and functional outcome of operative management of traumatic spinal cord injury patients admitted to a tertiary care centre in Northeast India., Methods: thirty patients with spinal cord injury admitted to a tertiary care centre from December 2019 to November 2021, and treated with instrumented stabilisation for spinal cord injury were evaluated until 6 months postoperatively. Patients were evaluated with validated neurological (American Spinal Injury Association scale) and functional outcome measures (Barthel index). Demographic details, mode of injury, morphology, patterns of fractures, neurological level, and management methods in the hospital were recorded and analysed using the Statistical Package for the Social Science (SPSS) version 27.0., Results: thoracolumbar spinal cord was more commonly injured with 16 (53.3%) patients compared to cervical spinal cord injury patients at 14 (46.7%). Eight patients had complete recovery, 7 patients had incomplete recovery and 15 patients had no recovery. At 6 months post-injury, 18 (60%) patients had favourable functional outcome. American Spinal Injury Association (ASIA) grade at admission was found to be significantly associated with the functional outcome., Conclusion: after surgery half of the patients had an improvement in their neurology, and functional outcome was favorable which suggests that surgery still holds the key to a better functional and rehabilitation outcome., Competing Interests: The authors declare no competing interests., (Copyright: Tuhin Purkayastha et al.)

This study aims to characterize the cytomorphology and ultrastructure of blood cells of native cattle of Mizoram. Twelve numbers of blood samples (10 ml) were collected from the Zobawng cattle, irrespective of sex. Blood smears were prepared and stained with different stains for cytomorphological study. The standard protocol has been followed for preparing blood samples for electron microscopy. Under a light microscope, erythrocytes of cattle were non-nucleated and round. The neutrophils were round, and the cytoplasm contained cytoplasmic granules. The eosinophils were rounded in outline with distinct cytoplasmic granules. The presence of basophils was infrequent with distinct blue color cytoplasmic granules. Small, medium, and large types of lymphocytes were recorded. The monocytes were round to oval in outline. Platelets were irregular to round. The reticulocytes were recorded occasionally, like small blue thin rods or granules. The cytoplasm and nucleus of granulocytes fluoresced greenish-yellow and red, respectively, with supravital stain. Under scanning electron microscopy (SEM), the erythrocytes appeared as biconcave discs. Different leukocytes were observed with their finger-like, plate-like, and narrow cell processes on their surface. Platelets were irregular structures. In transmission electron microscopy (TEM), the erythrocytes appeared anucleated biconcave elongated, neutrophils were roughly rounded with small cytoplasmic processes, and eosinophils were roughly circular with small cytoplasmic processes, the basophils were roughly circular with oval to elongated cytoplasmic granules, lymphocytes were roughly circular with centrally placed well-marked oval indented nucleus and some cytoplasmic processes, monocytes appeared spherical with long thick cytoplasmic processes and the non-nucleated platelets appeared roughly round to elongated. RESEARCH HIGHLIGHTS: Blood tests are a very good tool for determining animal health and diagnosing hematological illnesses. This study showed the cytomorphological and ultrastructural features of the blood cells of native cattle (Zobawng) of Mizoram state of India. The cytomorphological studies revealed that the morphological features of the blood cells of Zobawng cattle resembled to other domestic and wild animals, however, there were some differences in the cellular components. Nuclear DNA appeared orthochromatically greenish-yellow, but cytoplasmic RNA was observed to be metachromatically red under the fluorescence microscope. scanning electron microscopic (SEM) and transmission electron microscopic (TEM) results revealed that the ultrastructural features of the blood cells of Zobawng cattle contain certain cellular morphological differences as compared to the other domestic and wild animals., (© 2022 Wiley Periodicals LLC.)

Introduction: We aimed to examine how public health policies influenced the dynamics of coronavirus disease 2019 (COVID-19) time-varying reproductive number ( R t ) in South Carolina from February 26, 2020, to January 1, 2021., Methods: COVID-19 case series (March 6, 2020, to January 10, 2021) were shifted by 9 d to approximate the infection date. We analyzed the effects of state and county policies on R t using EpiEstim. We performed linear regression to evaluate if per-capita cumulative case count varies across counties with different population size., Results: R t shifted from 2-3 in March to <1 during April and May. R t rose over the summer and stayed between 1.4 and 0.7. The introduction of statewide mask mandates was associated with a decline in R t (-15.3%; 95% CrI, -13.6%, -16.8%), and school re-opening, an increase by 12.3% (95% CrI, 10.1%, 14.4%). Less densely populated counties had higher attack rates ( P < 0.0001)., Conclusions: The R t dynamics over time indicated that public health interventions substantially slowed COVID-19 transmission in South Carolina, while their relaxation may have promoted further transmission. Policies encouraging people to stay home, such as closing nonessential businesses, were associated with R t reduction, while policies that encouraged more movement, such as re-opening schools, were associated with R t increase.

Although prokaryotic DNA methylation investigations have long focused on immunity against exogenous DNA, it has been recently recognized that DNA methylation impacts gene expression and phase variation in Streptococcus pneumoniae and Streptococcus suis. A comprehensive analysis of DNA methylation is lacking for beta-hemolytic streptococci, and thus we sought to examine DNA methylation in the major human pathogen group A Streptococcus (GAS). Using a database of 224 GAS genomes encompassing 80 emm types, we found that nearly all GAS strains encode a type I restriction modification (RM) system that lacks the hsdS' alleles responsible for impacting gene expression in S. pneumoniae and S. suis. The GAS type I system is located on the core chromosome, while sporadically present type II orphan methyltransferases were identified on prophages. By combining single-molecule real-time (SMRT) analyses of 10 distinct emm types along with phylogenomics of 224 strains, we were able to assign 13 methylation patterns to the GAS population. Inactivation of the type I RM system, occurring either naturally through phage insertion or through laboratory-induced gene deletion, abrogated DNA methylation detectable via either SMRT or MinION sequencing. Contrary to a previous report, inactivation of the type I system did not impact transcript levels of the gene ( mga ) encoding the key multigene activator protein (Mga) or Mga-regulated genes. Inactivation of the type I system significantly increased plasmid transformation rates. These data delineate the breadth of the core chromosomal type I RM system in the GAS population and clarify its role in immunity rather than impacting Mga regulon expression. IMPORTANCE The advent of whole-genome approaches capable of detecting DNA methylation has markedly expanded appreciation of the diverse roles of epigenetic modification in prokaryotic physiology. For example, recent studies have suggested that DNA methylation impacts gene expression in some streptococci. The data described herein are from the first systematic analysis of DNA methylation in a beta-hemolytic streptococcus and one of the few analyses to comprehensively characterize DNA methylation across hundreds of strains of the same bacterial species. We clarify that DNA methylation in group A Streptococcus (GAS) is primarily due to a type I restriction modification (RM) system present in the core genome and does not impact mga -regulated virulence gene expression, but does impact immunity against exogenous DNA. The identification of the DNA motifs recognized by each type I RM system may assist with optimizing methods for GAS genetic manipulation and help us understand how bacterial pathogens acquire exogenous DNA elements.

Clonal replacement is a major driver for changes in bacterial disease epidemiology. Recently, it has been proposed that episodic emergence of novel, hypervirulent clones of group A Streptococcus (GAS) results from acquisition of a 36-kb DNA region leading to increased expression of the cytotoxins Nga (NADase) and SLO (streptolysin O). We previously described a gene fusion event involving the gene encoding the GAS M protein ( emm ) and an adjacent M-like protein ( enn ) in the emm4 GAS population, a GAS emm type that lacks the hyaluronic acid capsule. Using whole-genome sequencing of a temporally and geographically diverse set of 1,126 isolates, we discovered that the North American emm4 GAS population has undergone clonal replacement with emergent GAS strains completely replacing historical isolates by 2017. Emergent emm4 GAS strains contained a handful of small genetic variations, including the emm-enn gene fusion, and showed a marked in vitro growth defect compared to historical strains. In contrast to other previously described GAS clonal replacement events, emergent emm4 GAS strains were not defined by acquisition of exogenous DNA and had no significant increase in transcript levels of nga and slo toxin genes via RNA sequencing and quantitative real-time PCR analysis relative to historic strains. Despite the in vitro growth differences, emergent emm4 GAS strains were hypervirulent in mice and ex vivo growth in human blood compared to historical strains. Thus, these data detail the emergence and dissemination of a hypervirulent acapsular GAS clone defined by small, endogenous genetic variation, thereby defining a novel model for GAS strain replacement. IMPORTANCE Severe invasive infections caused by group A Streptococcus (GAS) result in substantial morbidity and mortality in children and adults worldwide. Previously, GAS clonal strain replacement has been attributed to acquisition of exogenous DNA leading to novel virulence gene acquisition or increased virulence gene expression. Our study of type emm4 GAS identified emergence of a hypervirulent GAS clade defined by variation in endogenous DNA content and lacking augmented toxin gene expression relative to replaced strains. These findings expand our understanding of the molecular mechanisms underlying bacterial clonal emergence.

Catabolite control protein A (CcpA) is a master regulator of carbon source utilization and contributes to the virulence of numerous medically important Gram-positive bacteria. Most functional assessments of CcpA, including interaction with its key co-factor HPr, have been performed in nonpathogenic bacteria. In this study we aimed to identify the in vivo DNA binding profile of CcpA and assess the extent to which HPr is required for CcpA-mediated regulation and DNA binding in the major human pathogen group A Streptococcus (GAS). Using a combination RNAseq/ChIP-seq approach, we found that CcpA affects transcript levels of 514 of 1667 GAS genes (31%) whereas direct DNA binding was identified for 105 GAS genes. Three of the directly regulated genes encode the key GAS virulence factors Streptolysin S, PrtS (IL-8 degrading proteinase), and SpeB (cysteine protease). Mutating CcpA Val301 to Ala (strain 2221-CcpA-V301A) abolished interaction between CcpA and HPr and impacted the transcript levels of 205 genes (40%) in the total CcpA regulon. By ChIP-seq analysis, CcpAV301A bound to DNA from 74% of genes bound by wild-type CcpA, but generally with lower affinity. These data delineate the direct CcpA regulon and clarify the HPr-dependent and independent activities of CcpA in a key pathogenic bacterium., (© 2020 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.)

To investigate the complex Pb-Cd exposure network in school-going children, a thorough investigation of the probable exposure means (diet, water, and school micro-environments such as paint dust and school courtyard soil) and exposure route (ingestion, inhalation and dermal) was carried out in a periurban area spanning three districts in southern Assam, India. Multivariate statistical analysis was carried out to understand the complex data matrices, and the health risk assessments (carcinogenic and non-carcinogenic) based on US EPA Risk Assessment models were also made. We found the median values to be 0.9-4.0 mg Pb/kg and 0.21-6.2 mg Cd/kg in various food items. Groundwater also had Pb (0.13-0.48 mg/L) and Cd (0.11-0.29 mg/L). Pb levels in paint dust were within the permissible limits, but 50% of the samples had higher than permissible levels of Cd. Approximately 23% of the school courtyard soil had Pb above the global background levels, but all the samples had 4-27 times elevated levels of Cd in them. School micro-environment contributed significantly to the metal load in children due to their typical hand-to-mouth behavior and dietary intake (food and water) via ingestion was the most prominent route of exposure in children. The evaluation of the estimated chronic daily intake and the hazard quotient indicated hazardous exposure over a lifetime to both Pb and Cd, but only Cd posed a prominent cancer risk. It could be concluded that chronic insidious effects of metals would be a noteworthy toxicological threat to children when exposed early on.

In this paper, a novel inter-layer exchange coupled (IEC) based 3-input full adder design methodology is proposed and subsequently the architecture has been implemented on the widely accepted micromagnetic OOMMF platform. The impact of temperature on the IEC coupled full-adder design has been analyzed up to Curie temperature. It was observed that even up to Curie temperature the IEC based adder design was able to operate at sub-50 nm as contrast to dipole coupled adder design which failed at 5 K for sub 50 nm. Simulation results obtained from OOMMF micromagnetic simulator shows, the IEC based adder design was at a lower energy state as compared to the dipole coupled adder indicating a more stable system and as the temperature of the design was increased, the total energy increased resulting in reduced stability. Potential explanation for the thermodynamic stability of IEC model lies in its energetically favored architecture, such that the total energy was lower than its dipole coupled counterparts. IEC architecture demonstrates supremacy in reliability and strength enabling NML to march towards beyond CMOS devices., (© 2021 IOP Publishing Ltd.)

In this paper, we propose an interlayer exchange coupling (IEC) based 3D universal NAND/NOR gate design methodology for the reliable and robust implementation of nanomagnetic logic design as compared to the state-of-the art architectures. Owing to stronger coupling scheme as compared to the conventional dipole coupling, the random flip of the states of the nanomagnets (i.e. the soft error) is reduced resulting in greater scalability and better data retention at the deep sub-micron level. Results obtained from Object Oriented Micromagnetic Framework micromagnetic simulation show even at a Curie temperature of the nanomagnets coupled through IEC, the logic function works properly as opposed to dipole coupled nanomagnets which fails at 5 K when scaled down to sub 50 nm. Contemplating the fabrication challenges, the robustness of the IEC design was studied for structural defects, positional misalignment, shape, and size variations. This proposed 3D universal gate design methodology benefits from the miniaturization of nanomagnets as well as reduces the effect of thermally induced errors resulting in opening up a new perspective for nanomagnet based design in magneto-logic devices.

Graphene interconnects have been projected to out-perform Copper interconnects in the next generation Magnetic Quantum-dot Cellular Automata (MQCA) based nano-electronic applications. In this paper a simple two-step lithography process for patterning CVD monolayer graphene on SiO 2 /Si substrate has been used that resulted in the current density of one order higher magnitude as compared to the state-of-the-art graphene-based interconnects. Electrical performances of the fabricated graphene interconnects were evaluated, and the impact of temperature and size on the current density and reliability was investigated. The maximum current density of 1.18 ×10 8 A/cm 2 was observed for 0.3 μm graphene interconnect on SiO 2 /Si substrate, which is about two orders and one order higher than that of conventionally used copper interconnects and CVD grown graphene respectively, thus demonstrating huge potential in outperforming copper wires for on-chip clocking. The drop in current at 473 K as compared to room temperature was found to be nearly 30%, indicating a positive temperature coefficient of resistivity (TCR). TCR for all cases were studied and it was found that with decrease in width, the sensitivity of temperature also reduces. The effect of resistivity on the breakdown current density was analysed on the experimental data using Matlab and found to follow the power-law equations. The breakdown current density was found to have a reciprocal relationship to graphene interconnect resistivity suggesting Joule heating as the likely mechanism of breakdown.

Hepatitis E virus (HEV) infection causes chronic hepatitis in solid organ transplant (SOT) recipients. Antiviral therapy consists of three months of ribavirin, although response rates are not optimal. We characterized plasma HEV kinetic patterns in 41 SOT patients during ribavirin therapy. After a median pharmacological delay of three (range: 0-21) days, plasma HEV declined from a median baseline level of 6.12 (3.53-7.45) log copies/mL in four viral kinetic patterns: (i) monophasic ( n = 18), (ii) biphasic ( n = 13), (iii) triphasic ( n = 8), and (iv) flat-partial response ( n = 2). The mean plasma HEV half-life was estimated to be 2.0 ± 0.96 days. Twenty-five patients (61%) had a sustained virological response (SVR) 24 weeks after completion of therapy. Viral kinetic patterns (i)-(iii) were not associated with baseline characteristics or outcome of therapy. A flat-partial response was associated with treatment failure. All patients with a log concentration decrease of plasma HEV at day seven of >15% from baseline achieved SVR. In conclusion, viral kinetic modeling of plasma HEV under ribavirin therapy showed, for the first time, four distinct kinetic profiles, a median pharmacologic delay of three days, and an estimated HEV half-life of two days. Viral kinetic patterns were not associated with response to therapy, with the exception of a flat-partial response.

Inactivating mutations in the control of virulence two-component regulatory system (covRS) often account for the hypervirulent phenotype in severe, invasive group A streptococcal (GAS) infections. As CovR represses production of the anti-phagocytic hyaluronic acid capsule, high level capsule production is generally considered critical to the hypervirulent phenotype induced by CovRS inactivation. There have recently been large outbreaks of GAS strains lacking capsule, but there are currently no data on the virulence of covRS-mutated, acapsular strains in vivo. We investigated the impact of CovRS inactivation in acapsular serotype M4 strains using a wild-type (M4-SC-1) and a naturally-occurring CovS-inactivated strain (M4-LC-1) that contains an 11bp covS insertion. M4-LC-1 was significantly more virulent in a mouse bacteremia model but caused smaller lesions in a subcutaneous mouse model. Over 10% of the genome showed significantly different transcript levels in M4-LC-1 vs. M4-SC-1 strain. Notably, the Mga regulon and multiple cell surface protein-encoding genes were strongly upregulated-a finding not observed for CovS-inactivated, encapsulated M1 or M3 GAS strains. Consistent with the transcriptomic data, transmission electron microscopy revealed markedly altered cell surface morphology of M4-LC-1 compared to M4-SC-1. Insertional inactivation of covS in M4-SC-1 recapitulated the transcriptome and cell surface morphology. Analysis of the cell surface following CovS-inactivation revealed that the upregulated proteins were part of the Mga regulon. Inactivation of mga in M4-LC-1 reduced transcript levels of multiple cell surface proteins and reversed the cell surface alterations consistent with the effect of CovS inactivation on cell surface composition being mediated by Mga. CovRS-inactivating mutations were detected in 20% of current invasive serotype M4 strains in the United States. Thus, we discovered that hypervirulent M4 GAS strains with covRS mutations can arise in an acapsular background and that such hypervirulence is associated with profound alteration of the cell surface., Competing Interests: The authors have declared that no competing interests exist.

In this paper, we report on the interesting phenomenon of magnetic phase transitions (MPTs) observed under the combined influence of an electric field (E) and temperature (T) leading to a thermo-electromagnetic effect on the pristine single-layer zigzag graphene nanoribbon (szGNR). Density functional theory-based first principles calculations have been deployed for this study on the intrinsic magnetic properties of graphene. Interestingly, by tuning electric field (E) and temperature (T), three distinct magnetic phase behaviors, para-, ferro- and antiferromagnetic are exhibited in pristine szGNR. We have investigated the unrivaled positional parameters of these MPTs. MPT occurring in the system also follows a positional trend and the change in these positional parameters with regard to the size of the szGNR along with the varied E and T is studied. We propose a bow-tie schematic to induce the intrinsic magnetism in graphene and present the envisaged model of the processor application with the reported intrinsic MPT in szGNR. This fundamental insight into the intrinsic MPTs in graphene is an essential step towards developing graphene-based spin-transfer torque magnetoresistive random access memory, quantum computing devices, magnonics and spintronic memory application.

Group A Streptococcus (GAS) is classified on the basis of the sequence of the gene encoding the M protein (emm) and the patterns into which emm types are grouped. We discovered a novel emm pattern in emm4 GAS, historically considered pattern E, arising from a fusion event between emm and the adjacent enn gene. We identified the emm-enn fusion event in 51 out of 52 emm4 GAS strains isolated by national surveillance in 2015. GAS isolates with an emm-enn fusion event completely replaced pattern E emm4 strains over a 4-year span in Houston (2013-2017). The novel emm-enn gene fusion and new emm pattern has potential vaccine implications.

The control of virulence regulator/sensor kinase (CovRS) two-component system is critical to the infectivity of group A streptococcus (GAS), and CovRS inactivating mutations are frequently observed in GAS strains causing severe human infections. CovS modulates the phosphorylation status and with it the regulatory effect of its cognate regulator CovR via its kinase and phosphatase activity. However, the contribution of each aspect of CovS function to GAS pathogenesis is unknown. We created isoallelic GAS strains that differ only by defined mutations which either abrogate CovR phosphorylation, CovS kinase or CovS phosphatase activity in order to test the contribution of CovR phosphorylation levels to GAS virulence, emergence of hypervirulent CovS-inactivated strains during infection, and GAS global gene expression. These sets of strains were created in both serotype M1 and M3 backgrounds, two prevalent GAS disease-causing serotypes, to ascertain whether our observations were serotype-specific. In both serotypes, GAS strains lacking CovS phosphatase activity (CovS-T284A) were profoundly impaired in their ability to cause skin infection or colonize the oropharynx in mice and to survive neutrophil killing in human blood. Further, response to the human cathelicidin LL-37 was abrogated. Hypervirulent GAS isolates harboring inactivating CovRS mutations were not recovered from mice infected with M1 strain M1-CovS-T284A and only sparsely recovered from mice infected with M3 strain M3-CovS-T284A late in the infection course. Consistent with our virulence data, transcriptome analyses revealed increased repression of a broad array of virulence genes in the CovS phosphatase deficient strains, including the genes encoding the key anti-phagocytic M protein and its positive regulator Mga, which are not typically part of the CovRS transcriptome. Taken together, these data establish a key role for CovS phosphatase activity in GAS pathogenesis and suggest that CovS phosphatase activity could be a promising therapeutic target in GAS without promoting emergence of hypervirulent CovS-inactivated strains., Competing Interests: The authors have declared that no competing interests exist.

Background: Pathobionts, bacteria that are typically human commensals but can cause disease, contribute significantly to antimicrobial resistance. Staphylococcus epidermidis is a prototypical pathobiont as it is a ubiquitous human commensal but also a leading cause of healthcare-associated bacteremia. We sought to determine the etiology of a recent increase in invasive S. epidermidis isolates resistant to linezolid., Methods: Whole-genome sequencing (WGS) was performed on 176 S. epidermidis bloodstream isolates collected at the MD Anderson Cancer Center in Houston, Texas, between 2013 and 2016. Molecular relationships were assessed via complementary phylogenomic approaches. Abundance of the linezolid resistance determinant cfr was determined in stool samples via reverse-transcription quantitative polymerase chain reaction., Results: Thirty-nine of the 176 strains were linezolid resistant (22%). Thirty-one of the 39 linezolid-resistant S. epidermidis infections were caused by a particular clone resistant to multiple antimicrobials that spread among leukemia patients and carried cfr on a 49-kb plasmid (herein called pMB151a). The 6 kb of pMB151a surrounding the cfr gene was nearly 100% identical to a cfr-containing plasmid isolated from livestock-associated staphylococci in China. Analysis of serial stool samples from leukemia patients revealed progressive staphylococcal domination of the intestinal microflora and an increase in cfr abundance following linezolid use., Conclusions: The combination of linezolid use plus transmission of a multidrug-resistant clone drove expansion of invasive, linezolid-resistant S. epidermidis. Our results lend support to the notion that a combination of antibiotic stewardship plus infection control measures may help to control the spread of a multidrug-resistant pathobiont.

Enterococcus faecalis is paradoxically a dangerous nosocomial pathogen and a normal constituent of the human gut microbiome, an environment rich in ethanolamine. E. faecalis carries the eut (ethanolamine utilization) genes, which enable the catabolism of ethanolamine (EA) as a valuable source of carbon and/or nitrogen. EA catabolism was previously shown to contribute to the colonization and growth of enteric pathogens, such as Salmonella enterica serovar Typhimurium and enterohemorrhagic Escherichia coli (EHEC), in the gut environment. We tested the ability of eut mutants of E. faecalis to colonize the gut using a murine model of gastrointestinal (GI) tract competition and report the surprising observation that these mutants outcompete the wild-type strain. IMPORTANCE Some bacteria that are normal, harmless colonizers of the human body can cause disease in immunocompromised patients, particularly those that have been heavily treated with antibiotics. Therefore, it is important to understand the factors that promote or negate these organisms' ability to colonize. Previously, ethanolamine, found in high concentrations in the GI tract, was shown to promote the colonization and growth of bacteria associated with food poisoning. Here, we report the surprising, opposite effect of ethanolamine utilization on the commensal colonizer E. faecalis , namely, that loss of this metabolic capacity made it a better colonizer., (Copyright © 2018 Kaval et al.)

Objectives: Neglected tropical diseases (NTD), account for a large proportion of the global disease burden, and their control faces several challenges including diminishing human and financial resources for those distressed from such diseases. Visceral leishmaniasis (VL), the second-largest parasitic killer (after malaria) and an NTD affects poor populations and causes considerable cost to the affected individuals. Mathematical models can serve as a critical and cost-effective tool for understanding VL dynamics, however, complex array of socio-economic factors affecting its dynamics need to be identified and appropriately incorporated within a dynamical modeling framework. This study reviews literature on vector-borne diseases and collects challenges and successes related to the modeling of transmission dynamics of VL. Possible ways of creating a comprehensive mathematical model is also discussed., Methods: Published literature in three categories are reviewed: (i) identifying non-traditional but critical mechanisms for VL transmission in resource limited regions, (ii) mathematical models used for dynamics of Leishmaniasis and other related vector borne infectious diseases and (iii) examples of modeling that have the potential to capture identified mechanisms of VL to study its dynamics., Results: This review suggests that VL elimination have not been achieved yet because existing transmission dynamics models for VL fails to capture relevant local socio-economic risk factors. This study identifies critical risk factors of VL and distribute them in six categories (atmosphere, access, availability, awareness, adherence, and accedence). The study also suggests novel quantitative models, parts of it are borrowed from other non-neglected diseases, for incorporating these factors and using them to understand VL dynamics and evaluating control programs for achieving VL elimination in a resource-limited environment., Conclusions: Controlling VL is expensive for local communities in endemic countries where individuals remain in the vicious cycle of disease and poverty. Smarter public investment in control programs would not only decrease the VL disease burden but will also help to alleviate poverty. However, dynamical models are necessary to evaluate intervention strategies to formulate a cost-effective optimal policy for eradication of VL.

Catabolite control protein A (CcpA) is a highly conserved, master regulator of carbon source utilization in gram-positive bacteria, but the CcpA regulon remains ill-defined. In this study we aimed to clarify the CcpA regulon by determining the impact of CcpA-inactivation on the virulence and transcriptome of three distinct serotypes of the major human pathogen Group A Streptococcus (GAS). CcpA-inactivation significantly decreased GAS virulence in a broad array of animal challenge models consistent with the idea that CcpA is critical to gram-positive bacterial pathogenesis. Via comparative transcriptomics, we established that the GAS CcpA core regulon is enriched for highly conserved CcpA binding motifs (i.e. cre sites). Conversely, strain-specific differences in the CcpA transcriptome seems to consist primarily of affected secondary networks. Refinement of cre site composition via analysis of the core regulon facilitated development of a modified cre consensus that shows promise for improved prediction of CcpA targets in other medically relevant gram-positive pathogens.