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KATP channel gain- (GOF) and loss-of-function (LOF) mutations in SUR1 or Kir6.2 underlie human neonatal diabetes mellitus (NDM) and congenital hyperinsulinism (CHI), respectively. Genetically modified mice with transgenic overexpression of GOF mutations recapitulate many features of human NDM and transgenic mice expressing incomplete KATP LOF do reiterate mild hyperinsulinism, although complete KATP knockout mice do not exhibit persistent hyperinsulinism. We have shown that islet excitability and glucose homeostasis in zebrafish are regulated by identical KATP channels to those in mammals, and to explore the possibility of using zebrafish for modeling CHI, we have examined SUR1 truncation mutation (K499X) introduced into the abcc8 gene. Patch-clamp analysis confirmed complete absence of channel activity in β-cells from K499X (SUR1-/-) fish. No difference in random blood glucose was detected in heterozygous SUR1+/- fish, nor in homozygous SUR1-/- fish, mimicking findings in SUR1 knockout mice. Mutant fish also demonstrated impaired glucose tolerance, similar to LOF mouse models. In paralleling features of mammalian hyperinsulinism resulting from loss-of-function mutations, this gene-edited animal provides a valid zebrafish model of KATP LOF driven-dependent pancreatic disease.Key PointsLoss-of function in the Kir6.2 (KCNJ11) and SUR1 (ABCC8)-encoded pancreatic islet β-cell KATP channels underlie congenital hyperinsulinism. Mouse models reiterate key features, but zebrafish models could provide a powerful model for further analysis and therapy testing.An early nonsense mutation in exon 10 of SUR1 was generated by ENU mutagenesis.Patch-clamp analysis revealed an absence of β-cells of SUR1 truncation mutants.Ca imaging demonstrated elevated basal [Ca]i in β-cells with SUR1 truncation.Homozygous SUR1 truncation mutants had normal fasting glucose but impaired glucose tolerance as adults, mimicking findings in mouse SUR1 knockouts.In paralleling features of mammalian diabetes and hyperinsulinism resulting from equivalent loss-of-function mutations, this gene-edited animal provides a valid zebrafish model of KATP -LOF dependent pancreatic diseases.

This paper documents the auditing profession’s gender salary gap in a setting where the overall society consists of a high degree of gender equality. Using Swedish administrative data from 2007 to 2015 for all CPAs, I find that the auditing profession’s overall gender salary gap has substantially declined: the gap in 2015 is less than half the size of the 2007 gap. More female auditors have moved up to the top earnings group during this period, demonstrating an evident lessening of the glass ceiling phenomenon. Oaxaca-Blinder analysis shows that auditor’s clientele size is the most important factor in explaining the salary gap, and the increase in female auditors’ clientele size during this period accounts for half of the decrease in the total salary gap. Further, I also find evidence that the rise of female leadership in Big Six firms is positively associated with the increase in female auditors’ clientele size. Consistent with Kunze and Miller (2017), such ‘women help women’ effect in the auditing environment is more pronounced in the middle and lower half of the firm hierarchy, suggesting that female representation at the top of the firm has spillover benefits for lower-ranked female auditors. The implications of this study may help audit firms close their gender gap and better cope with the overall auditing industry’s talent challenges.

Background The exogenous application of priming molecules to plants helps them to develop tolerance against salinity stress. In the present study, we used exogenous naringenin (0.5 mM) pretreatment before the stress in safflower seedlings under 25 mM NaCl to elucidate the role of naringenin to alleviate oxidative conditions associated with salinity complications. Results Our results showed biomass, leaf relative water content, chlorophyll content, K+ content, and K+/Na+ ratio were negatively affected by 25 mM NaCl. However, the H2O2 accumulation, malondialdehyde (MDA) content, antioxidant enzymes and Na+ content of NaCl-stressed safflower seedlings were remarkably increased. The results obtained in the present study showed the beneficial effects of the pre-treatment of naringenin in safflower seedlings under non-salinity stress condition with respect to increasing plant biomass, total phenolic compound, radical scavenging activity (RSA), soluble sugar content, proline, glutathione, enzymatic antioxidants, and K+ content. Also, the results showed that naringenin pre-treatment can (partly) be overcome NaCl-induced stress on safflower seedlings, probably due to higher accumulation of plant biomass, total phenolic compound, RSA, catalase (CAT) activity, and K+/Na+ ratio as well as lowering the H2O2 and MDA content in the leaves. Conclusions Generally, it could be concluded that, pre-treatment of naringenin before stress could partly diminish NaCl-caused oxidative stress in safflower seedlings, probably due to improvement in enzymatic and non-enzymatic antioxidant and reduced cell membrane damage.

Objective: As medical cannabis use increases in North America, establishing the safety profile of cannabis is a priority. The objective of this study was to assess rates of emergency department (ED) visits and hospitalizations due to poisoning by cannabis and cannabis-related mental health disorders among medically authorized cannabis patients in Ontario, Canada, between 2014 and 2017.Methods: This is a retrospective longitudinal study conducted among a cohort of patients who received an authorization to use cannabis for treating various health conditions in Ontario, Canada. The cannabis cohort was selected using data collected in participating Canadian cannabis clinics. Outcomes included: ED visit/hospitalization with a main diagnosis code for cannabis/cannabinoid poisoning; and ED visit/hospitalization with a main diagnosis code for mental/behavioural disorders due to cannabis use. The Spearman correlation and univariate Cox proportional hazard regression was utilized.Results: From 29153 patients who received medical authorization, 23091 satisfied the inclusion criteria. During a median follow-up of 240 days, 14 patients visited the ED or were hospitalized for cannabis poisoning – with an incidence rate of 8.06 per 10,000 person-years patients (95%CI: 4.8-13.6). A total of 26 patients visited the ED or were hospitalized for mental and behavioural disorders due to cannabis use- with an incidence rate of 15.0 per 10,000 person-years (95%CI: 10.2-22.0). Predictors of cannabis-related mental and behavioural disorders include prior substance use disorders (drugs and alcohol), other mental disorders, age, diabetes, and chronic obstructive pulmonary disease. Conclusions: The results suggest that the incidence of cannabis poisoning or cannabis-related mental and behavioural disorders was low among patients who were authorized to use cannabis to treat a health condition. Identified predictors can help to target patients with potential risk of the studied outcomes.

The study is to evaluate the impact of COVID-19 in the Pediatric Oncology Units (POUs) of Pakistan. Data from 1 April 2019 to 30 June 2019 and 1 April 2020 to 30 June 2020 for the first and second cohort, respectively, in order to compare the registration, abandonment rate, and delay in treatment. Six hundred and thirty-four were registered cases, 379 and 255 in the first and second cohort, respectively, which was significantly different0.005. Seventy-seven were abandoned, 45 and 32 in the first and second cohort, respectively. Fifty-nine COVID-19 positive cases, 24, 4, 27, and 4 were admitted, referred, home isolated, and leave against medical advice (LAMA), respectively. Delayed treatment and reduction in new cases were observed.

Background Adequate nutritional intake plays a pivotal role in optimising performance, recovery and body composition goals. This study aimed to investigate the dietary intakes (DI), nutritional knowledge (NK) and attitudes, perceptions and challenges (APC) of semi-professional rugby players in Scotland. Methods Dietary intakes and NK of 24 male semi-professional rugby players of a Super6 club were evaluated using validated questionnaires. Players were categorized as having good or poor NK according to NK scores. Diet-related APCs were assessed online using researcher developed questionnaires and 1–1 semi-structured interviews. Results Mean ± SD total NK% was poor, 53.7 ± 11.9%. The ‘Good’ NK group scored significantly higher in the Weight Management (p = 0.014), Macronutrients (p

We aimed to analyze whether rapid weight gain (RWG) between the official weigh-in and the time of the fight was associated with fight success in MMA. A total of 700 professional MMA fights involving 1,400 weigh-ins from 21 MMA promotions regulated by the California State Athletic Commission were analyzed. Multilevel logistic regression accounting for individual (i.e., athlete) and cluster levels (i.e., fights) was used to analyze the association of all measures with a theoretical relationship with the dependent variable and without interdependency with one another (i.e., %RWG, sex, body mass division, competition level) with the fight outcome (i.e., win or loss). The odds ratios (OR) with 95% confidence intervals (95%CI) were calculated. The highest mean %RWG was found for the flyweight, bantamweight, featherweight, and lightweight divisions. The %RWG significantly predicted the fight outcome (s=0.044; OR=1.045; 95%CI=1.014-1.078; p=0.005) so that for each 1% of additional RWG, the chance of winning increased by 4.5%. With the largest sample to date and in a 'real-world' scenario, the present results suggest that the magnitude of RWG is linked to the chance of winning in MMA combats. It is suggested that regulatory commissions, confederations, and event organizers should consider regulating RWG, considering that, despite its detrimental impact on the athletes' health and performance, the potential advantage might stimulate athletes to invest in rapid weight loss, followed by gain after the official weigh-in to increase their chance of winning.

As next-generation pathogen detection methods, CRISPR-Cas-based detection methods can perform single-nucleotide polymorphism (SNP) level detection with high sensitivity and good specificity. They do not require any particular equipment, which opens up new possibilities for the accurate detection and identification of Bacillus anthracis. In this study, we developed a complete detection system for B. anthracis based on Cas12a. We used two chromosomally located SNP targets and two plasmid targets to identify B. anthracis with high accuracy. The CR5 target is completely new. The entire detection process can be completed within 90 min without electrical power and with single-copy level sensitivity. We also developed an unaided-eye visualization system based on G4-DNAzyme for use with our CRISPR-Cas12a detection system. This visualization system has good prospects for deployment in field-based point-of-care detection. We used the antisense nucleic acid CatG4R as the detection probe, which showed stronger resistance to interference from components of the solution. CatG4R can also be designed as an RNA molecule for adaptation to Cas13a detection, thereby broadening the scope of the detection system.

The non-indigenous to the Mediterranean tropical seagrass Halophila stipulacea has the possibility to become more prevalent in the Mediterranean basin, exacerbated by the rapid increase of water temperature. Molecular profiling appears a promising tool to study the traits that render H. stipulacea tolerant and resilient and facilitate its rapid and vast geographical spread. Taking advantage from recent seagrass genomes sequencing, proteomics specialty has been applied to several seagrasses giving new insight on the biology and physiology of this group of angiosperms. Thus, it could be of interest to apply proteomics to H. stipulacea that it could be considered as a possible plant model species to study marine biological invasion. The first step to achieve this goal is to obtain high quality proteins from plant tissue. Tissue fixation and protein extraction protocol are the most challenging steps in proteomics. Here we report a fine-tuned procedure obtained by comparing protein yield from H. stipulacea plants frozen in liquid nitrogen or preserved in RNAlater and processed following two different extraction protocols. Higher protein yield have been extracted from the procedure that use the RNAlater preserved plants, extracted with trichloroacetic acid in water followed by trichloroacetic acid in acetone, compared to those obtained from all other procedures. Protein purity of these samples have been tested by the separation in SDS-PAGE comfirming a better resolved profile of peptide bands suitable for a gel-based proteomics. Then, to assess the quality of proteins the mHPLC-ESI-MS/MS mass spectrometry analyses and bioinformatics have been performed. Hundreds proteins have been identified against several seagrass genomic resources available at UniProt, NCBI, SeagrassDB and transcriptomic datasets, which were merged to form the first customized dataset useful for H. stipulacea proteomic investigations.

We highlight the need to include an analysis of the B-1 B cell subset to complement the characterization of the cell-mediated immune response to the Mycobacterium tuberculosis Beijing lineage. The literature describes the B-1 cell repertoire’s involvement in the cell-mediated response within granulomas, which is different from the classic antibody response B cells are generally associated with. Specifically, the B-1 B cell subset migrates from other compartments along with other cells to the infection site. We provide details to complement the reported results from Cerezo-Cortes et al.

Serum resistance is a poorly understood but common trait of some systemic disease pathogenic strains of bacteria. In this study, we analysed the role Escherichia coli type three secretion system 2 (ETT2) of avian pathogenic Escherichia coli (APEC) in serum resistance by bacteria survival number in serum culture, mRNA Seq and Tandem Mass Tag™ (TMT™) detection, lipopolysaccharide (LPS) extraction, and biofilm formation detection. We found that the ETT2 gene cluster deletion strain (ΔETT2) is more resistant to the killing effect of serum than wild type APEC40. The analysis of ΔETT2 compared to APEC40 in the transcriptomics and proteomics data showed that ETT2 has a negative effect in the ATP-binding cassette (ABC) translator system and quorum sensing system and a positive effect in purine metabolism. ETT2 may affect the LPS, biofilm, flagella, and fimbriae which may affect the serum resistance. These results could lead to effective strategies for managing the infection of APEC. The mRNA Seq data of this study are available in the Sequence Read Archive of the National Center for Biotechnology Information under the BioProject PRJNA757182, and proteomic raw data have been deposited under the accession number IPX0003420000 at iProX.

Animal models are the silent scouts that help to understand the complex biological processes and gather data that aid our understanding of how organisms function. Various animal models are being sacrificed to assess the impact of toxic chemicals. Mortality calculations should be minimized and much data should be collected on the basis of clinical signs that can contribute to identifying robust humane endpoints linked to mortality. This study was designed to calculate the lowest possible dose of PbAc (lead acetate), a neurotoxicant, that can have a toxicological impact on the zebrafish retina and to minimize animal usage. Dose and time-dependent changes were observed in the zebrafish retina following PbAc exposure with zero mortality. Vision and visual behavior response are the foremost indicators that can be recorded to mark the risk assessment of any chemical. Therefore, the present study aims at dose and time response to find the lowest dose of PbAc affecting the zebrafish retina and its visual behavior. Zebrafish were treated for 3 weeks with four concentrations of 0.04, 0.06, 0.08, and 0.1 mg/L of PbAc for a dose-response study. Then for the time response study, two doses 0.08 and 0.1 mg/L were selected and zebrafish were exposed to those concentrations for 2 and 4 weeks. The results of qualitative and quantitative analyses of retinal histology showed that 15 days of treatment with 0.08mg/L concentration can cause appropriate damage to the photoreceptor layer. At the ultrastructural level, it was further observed that PbAc induces damage to the photoreceptors, especially the rod cells. Escape response behavior showed a significant decrease in visual response to changing contrasts in an increasing dose-dependent manner.

Avian pathogenic Escherichia coli (APEC) is the bacterial pathogen of poultry colibacillosis, which causes significant economic losses to the poultry industry. The lack of an effective vaccine against multiple serotypes and the emergence of multi-resistant isolates have made the control of avian colibacillosis troublesome. To identify conserved potential vaccine candidates, 58 genomes of APEC were obtained (54 sequenced by our laboratory and 4 downloaded from NCBI). A reverse vaccinology (RV) method based on the pangenome - called Pan-RV analysis - was performed in APEC-protective protein mining for the first time. Finally, four proteins were selected, and their immunoreactivity with anti-O1, O2, and O78 serum was verified by western blotting. Our in silico method of analysis will pave the way for rapid screening of vaccine candidates and will lay the foundation for the development of a highly effective subunit vaccine controlling APEC infection.

Microorganisms mainly exist in the form of biofilm in nature. Biofilm can contaminate food and drinking water system, as well as cause chronic wound infections, thereby posing a potential threat to public health safety. In the last two decades, researchers have made efforts to investigate the genetic contributors control different stages of biofilm development (adherence, initiation, maturation, and dispersal). As an opportunistic pathogen, C. albicans causes severe superficial or systemic infections with high morbidity and mortality under conditions of immune dysfunction. It has been reported that 80% of C. albicans infections are directly or indirectly associated with biofilm formation on host or abiotic surfaces including indwelling medical devices, resulting in high morbidity and mortality. Significantly, the outcome of C. albicans biofilm development includes enhanced invasion, exacerbated inflammatory responses and intrinsic resistance to antimicrobial chemotherapy. Thus, this review aimed at providing a comprehensive overview of the regulatory network controls microbial biofilm development, with C. albicans as a representative, served as reference for therapeutic targets.

The physical demands of professional female football have intensified in recent years. Supplements are only advised in addition to a healthy, balanced diet, but may warrant a greater prevalence in the professional game to support well-being, recovery, and performance. Supplements used by players should be safe, legal, and scientifically proven to be effective. An individual approach should be taken to using supplements dependant on the needs and goals of the player. Female players should aim to improve the frequency of protein intake throughout the day, whilst tailoring doses to individual body mass. Vitamin D supplementation is vital throughout the winter months in countries with limited sun exposure, however doses should be administered based on individual blood test results. Iron is likely to be important to the well-being of female athletes throughout the season, in particular during the menses. Omega-3 and collagen may be of greater benefit to female than male athletes during recovery from soft tissue injury, whilst probiotics and creatine are beneficial throughout the season for reducing risk of illness and optimising recovery, respectively. Ergogenic supplements for football include beta-alanine, nitrate and caffeine. Caution should be taken with caffeine use due to the varying tolerance of difference athletes and sleep impairments that can follow.

Road networks can lead to serious negative impacts on wildlife populations. In particular, wildlife-vehicle collisions (WVC) caused by traffic is the best-known and visible effect of roads on wildl...

Microalgal cell attaching and biofilm formation are critical in the application of microalgal biocathode, which severs as one of the hopeful candidates to an original cathode in bioelectrochemical systems. Many efforts have been put in biofilm formation and bioelectrochemical systems for years, but the predominant factors shaping microalgal biocathode formation are sketchy. We launched a pair of researches to investigate microalgal attachment and biofilm formation in the presence/absence of applied voltages using Chlamydomonas microsphaera as a model unicellular motile microalga. In this study, we presented how microalga attached and biofilm formed on a carbon felt surface without applied voltages and try to manifest the most important aspects in this process. Results showed that while nutrient sources did not directly regulate cell attachment onto the carbon felt, limited initial nutrient concentration nevertheless promoted cell attachment. Specifically, nutrient availability did not influence the early stage (20–60 min) of microalgal cell attachment but did significantly impact cell attachment during later stages (240–720 min). Further analysis revealed that nutrient availability-mediated chemotactic movements and zeta potential are crucial to facilitate the initial attachment and subsequent biofilm formation of C. microsphaera onto the surfaces, serving as an important factor controlling microalgal surface attachment. Our results demonstrate that nutrient availability is a dominant factor controlling microalgal surface attachment and subsequent biofilm formation processes. This study provides a mechanistic understanding of microalgal surface attachment and biofilm formation processes on carbon felts surfaces in the absence of applied voltages.

In 2021, three encephalitis cases due to the Borna disease virus 1 (BoDV-1) were diagnosed in the north and east of Germany. The patients were from the states of Thuringia, Saxony-Anhalt, and Lower Saxony. All were residents of known endemic areas for animal Borna disease but without prior diagnosed human cases. Except for one recently detected case in the state of Brandenburg, all >30 notified cases had occurred in, or were linked to, the southern state of Bavaria. Of the three detected cases described here, two infections were acute, while one infection was diagnosed retrospectively from archived brain autopsy tissue samples. One of the acute cases survived, but is permanently disabled. The cases were diagnosed by various techniques (serology, molecular assays, and immunohistology) following a validated testing scheme and adhering to a proposed case definition. Two cases were classified as confirmed BoDV-1 encephalitis, while one case was a probable infection with positive serology and typical brain magnetic resonance imaging, but without molecular confirmation. Of the three cases, one full virus genome sequence could be recovered. Our report highlights the need for awareness of a BoDV-1 etiology in cryptic encephalitis cases in all areas with known animal Borna disease endemicity in Europe, including virus-endemic regions in Austria, Liechtenstein, and Switzerland. BoDV-1 should be actively tested for in acute encephalitis cases with residence or rural exposure history in known Borna disease-endemic areas.

Lassa fever (LF) is a deadly viral hemorrhagic disease that is endemic to West Africa. The causative agent of LF is Lassa virus (LASV), which causes approximately 300,000 infections and 5,000 deaths annually. There are currently no approved therapeutics or FDA-approved vaccines against LASV. The high genetic variability between LASV strains and immune evasion mediated by the virus complicate the development of effective therapeutics and vaccines. Here, we aim to provide a comprehensive review of the basic biology of LASV and its mechanisms of disease pathogenesis and virulence in various animal models, as well as an update on prospective vaccines, therapeutics, and diagnostics for LF. Until effective vaccines and/or therapeutics are available for use to prevent or treat LF, a better level of understanding of the basic biology of LASV, its natural genetic variations and immune evasion mechanisms as potential pathogenicity factors, and of the rodent reservoir-vector populations and their geographical distributions, is necessary for the development of accurate diagnostics and effective therapeutics and vaccines against this deadly human viral pathogen.

Type 2 diabetes mellitus (T2DM) is known as a complex genetic disease characterized by genetic and environmental factors. The imbalanced intestinal flora and intestinal mucosal barrier are considered to be related to T2DM. Curcumin has been proved to affect the progression of T2DM. T2DM animal was established by low-dose streptozotocin intraperitoneal injection combined with high-fat diet (HFD) feeding. Hematoxylin and eosin (HE) staining and transfer electron microscopy (TEM) were used to observe morphological changes of intestinal tissues of T2DM rats. Insulin and glucose tolerance tests were performed to investigate the influence of curcumin on blood glucose. Curcumin significantly improved the intestinal integrity, hyperglycemia and insulin resistance in diabetic rats. The metabolic endotoxemia induced by HFD in diabetic rats was inhibited remarkably. Curcumin reversed gut microbiota dysbiosis in diabetic rats caused by HFD. We demonstrated that curcumin could protect intestinal mucosal barrier, improve insulin resistance and reduce blood glucose in diabetic rats. This study might provide experimental evidence for the prevention and treatment in T2DM.

Purpose To study the distribution of 210Po concentrations in various parts of spice plants cultivated by conventional farming in Nilgiris, South India and estimate effective radiation dose received. Materials and methods Parts of plant such as leaf, fruit, seed, flower bud and rhizome of selected spice plants cultivated at various locations in the study area have been collected for estimation of 210Po concentration. Organic materials in known mass of processed samples have been removed by chemical method. 210Po concentration in the digested sample has been electrostatically deposited on pre-polished silver discs at constant temperature. ZnS(Ag) detector based alpha counting system with minimal detectable value 3.2 mBq has been used for estimation of 210Po activity concentration. Results Leaves of Curcuma longa have registered a high concentration of 45.6 ± 4.5 Bq kg-1 of 210Po and rhizome of Allium sativum has registered a low value of 4.7 ± 0.8 Bq kg-1. Leaves of spice plants have registered higher concentration of 210Po than their fruits, seeds, flower buds or rhizomes. Soil to plant transfer factor of 210Po has been found to vary from 0.13 for rhizome of A. sativum to 2.23 corresponding to leaf of C. longa. Concentration of 210Po estimated in leaves and mean leaf areas of spice plants have been found to have a correlation coefficient of 0.83, which indicates that absorption of 210Po deposited on surface area of leaves is the main source of this isotope in plant parts. Conclusions Consumption of various parts of spice plant does not pose significant radiological risk as effective dose received by the general public due to 210Po has been found to vary from 6.34 to 46.05 μSv y-1, less than the value recommended by ICRP.

1. The objective of this study was to examine the influence of dietary soluble non-starch polysaccharide (sNSP) level and xylanase supplementation on productive performance, viscosity and pH along the gastrointestinal tract in laying hens. Excreta moisture content, ileal and caecal microbiota and short-chain fatty acid (SCFA) composition and apparent total tract nutrient utilisation were measured.2. Hyline Brown laying hens (n = 144) were housed individually at 25 weeks of age and allocated to one of the four wheat-based dietary treatments in a 2 × 2 factorial arrangement, consisting of two levels of sNSP (High 13.40 g/kg or Low 11.22 g/kg), with or without xylanase (0 or 12,000 BXU/kg). Birds were fed the dietary treatments for 56 d.3. Increasing dietary sNSP increased jejunum viscosity, degradability of total NSP, total tract flow of insoluble arabinose, and succinic acid concentration in the caeca (P 0.05). Feeding high sNSP decreased excreta moisture content, total tract energy retention and free oligosaccharide, total tract flow of soluble and insoluble galactose and insoluble rhamnose and fucose, and ileal acetic and lactic acid concentrations (P 0.05), and tended to reduce egg production (P = 0.058).4. Supplementation with xylanase resulted in reduced jejunum and ileum viscosity, caecal pH, excreta moisture, flow of soluble arabinose and glucose and insoluble arabinose and xylose, caecal concentration of

Purpose To investigate the anti-inflammatory and anti-fungal role of α-melanocyte stimulating hormone (α-MSH) in Aspergillus Fumigatus (A. fumigatus) keratitis. Method Corneas of C57BL/6 mice were infected with A. Fumigatus. α-MSH (5 ul, 1x10-4mmol/ml) was given by subconjunctival injection from day 1 to day 3 post infection (p.i.). After 3 days p.i., clinical score was recored and HE staining was tested. Fungal load in mice corneas was observed by plate counting. Pro-inflammatory mediators and pattern recognition receptors (PRRs) were detected. The numbers of neutrophils and macrophages were tested by immunofluorescence staining. The role of α-MSH in RAW264.7 cells after A. fumigatus stimulation were evaluated by PCR and Western blot, and MPKA protein levels including total-JNK (T-JNK), phosphorylated-JNK (P-JNK), total-ERK (T-ERK) and phosphorylated-ERK (P-ERK) were tested via Western blot with or without α-MSH treatment. Results Compared with PBS control group, α-MSH treatment alleviated disease response and decreased clinical score at 3 days p.i. HE staining showed less infiltration in corneal tissue after α-MSH treatment. Plate counting experiment showed that number of viable fungus in corneas of α-MSH treated group was less than control group. mRNA levels of IL-1β, TNF-α, IL-6, MIP-2, LOX-1, Dectin-1 and iNOS were decreased. Protein levels of IL-1β, TNF-α, IL-6 and Dectin-1 were decreased. α-MSH treatment also decreased the infiltrating neutrophils and macrophages. The levels of pro-inflammatory cytokines, Dectin-1 and LOX-1 stimulated by A. fumigatus, were also suppressed by pretreatment of α-MSH in RAW264.7 cells. The ratio of P-JNK/T-JNK and P-ERK/T-ERK were down regulated in α-MSH group compared with PBS control group. Conclusion α-MSH alleviates the severity and decreases fungal load of A. fumigatus keratitis in mice. Migration of neutrophils and macrophages are restrained. α-MSH downregulates the expression of dectin-1 and the ratio of P-JNK/T-JNK and P-ERK/T-ERK in A. fumigatus infection.

Organelle-specific autophagy directs degradation of eukaryotic organelles under certain conditions. Like other organelles, peroxisomes are subject to autophagic turnover at lysosomes. However, peroxisome autophagy (pexophagy) has yet to be analyzed in a live-animal system, limiting knowledge on its regulation during an animal's life. Here, we generated a tandem-fluorophore reporter that enabled real-time tracking of pexophagy in live Caenorhabditis elegans. We observed that pexophagy occurred at a population of non-canonical, tubular lysosomes specifically during starvation and aging. Remarkably, in these contexts, tubular lysosomes were the predominant type of lysosome in the intestine, transforming from vesicles. Though we found that peroxisomes were largely eliminated in early adulthood, they appeared restored in new generations. We identified peroxisomal genes that regulated age-dependent peroxisome loss and demonstrated that modifying this process altered animal lifespan. These findings reveal new facets of peroxisome homeostasis relevant to aging and challenge the prevailing perception of lysosome homogeneity in autophagy.

Lilium regale Wilson (L.regale), originated in the Minjiang River basin in Sichuan, China, has different phenotypic characteristics in different environments. To analyze the correlation between the phenotypes of L.regale and its soil micro-ecological environment, wild habitat soil of L.regale at the two altitudes were selected to analyze the diversity and community structure of microorganisms in soil, and measure the soil physicochemical factors and enzyme activities. The structural composition and diversity of fungal and bacterial communities in hillside and valley soils were significantly different (p < 0.01). Soil available potassium (AK) and soil enzyme activities such as urease (S_UE), sucrase (S_SC), and catalase (S_CAT) differed significantly different between hillsides and valleys (p < 0.01), while organic matter (OM), total phosphorus (TP), and polyphenol oxidase (S_PPO) had no great variances. Correlation analysis was conducted between the common and differential microorganisms and the morphological characteristics, soil physicochemical factors and soil enzyme activities of L.regale in both hillside and valley. The results showed that both of the fungal and bacterial could be clustered into two distinct groups by positive and negative correlations, suggesting that the representative microorganism may have structural characteristics that are directly related to soil physicochemical properties and enzyme activities, which conversely affect the phenotype of Lily. Therefore, the study on the native species of horticultural plants and the local soil microhabitat environment will benefit the conservation of wild Lily and provide theoretical guidance for the domestication and breeding of horticultural plants.

Limited research exists on athletes' olfaction. As diet is an elementary part of an athlete's lifestyle and training, it is important to know whether the sport discipline itself carries a risk for olfactory impairment. The aim of this study was to evaluate whether elite swimmers (chemical irritation), boxers (head impact) or soccer players (head impact) are at risk for impaired olfaction. Elite-level male swimmers (n=30), boxers (n=35) and soccer players (n=30) aged 18-40 years were recruited from Finnish sport clubs. Floorball players (n=30) were recruited as a control group. All participants filled in a questionnaire about their training history, nasal and sinonasal diseases, asthma, nasal operations and traumas, smoking and self-evaluation of olfaction and taste function. Sniffin' Sticks odour identification test with 12 different odorants and anterior rhinoscopy were performed on all participants. The mean score from the smelling test did not differ between the sport groups. Sinonasal diseases and bronchial asthma were more common among swimmers than among the other athletes. Rhinitis symptoms were common among all athletes. Boxing, soccer or swimming does not seem to affect sense of smell. The majority of our participating elite athletes had normal olfaction, even if they had had a long history of active sports.

There is an ever-increasing range of antibiotic-resistant pathogens that have led to higher community-acquired infections, and substantial mortality rates in critically ill patients.We have critically appraised available evidence through a structured literature review, investigating effective empiric antibiotic administration and appropriateness on outcomes of critically ill patients with an increased risk of developing resistant pathogens. The use of new antibiotics should be determined based on relevant knowledge of their spectrum and properties to provide effective mode of action for critically ill patients.Restricting severely ill patients access to new broad-spectrum empirical drugs is not the answer. Rather there should be a focus on identifying host response to infection to differentiate between colonization or contamination and true infection, and the sensitivity to antibiotics used in the intensive care unit (ICU). Management relies on adequate antibiotic administration, the ability to monitor response, and facilitate the cessation of antibiotic treatment. The major determinant of patient success in a patient with a severe infection is the 'right' antibiotic or complementary course of treatment. As an overarching criterion, the following three appropriate "Ds" should be considered: Dosing, Duration, and De-escalation to empirically assess the right antibiotic optimal antimicrobial selection.

Forty-seven rock samples from the stratotypes of upper Paleogene formations - Prussian, Palve and Kurshskaya formations – are collected from the biggest amber deposit worldwide (Primorsky quarry, K...

H2 receptor antagonists are the medication given for treating stomach ulcers, but lately, reports have shown their role in healing several malignant ulcers. The present work entails the interaction of H2 blocker nizatidine with calf thymus (ct)-DNA for determining the binding mode and energetics of the interaction. Multi-spectroscopic, calorimetric, viscometric and bioinformatic analysis revealed that nizatidine interacted with ct-DNA via groove-binding mode and is characterised by exothermic reaction. Moreover, assessment of genotoxic potential of nizatidine in vitro was carried out in peripheral human lymphocytes by alkaline comet assay. DNA damage occurred at high concentrations of nizatidine. Genotoxicity of nizatidine was also evaluated in vivo by assessing cytogenetic biomarkers viz. micronuclei formation and chromosomal aberration test. Nizatidine was able to induce micronuclei formation and chromosomal damage at high dose. Additionally, cytotoxic activity of nizatidine was determined in cancer cell lines, namely HeLa and HCT-116 and compared with the normal human cell line HEK-293 employing MTT assay. It was observed that nizatidine was more toxic towards HeLa and HCT-116 than HEK-293. Cell morphology analysis by compound inverted microscopy further strengthens the finding obtained through MTT assay.

Natural product such as flavonoids and their derivatives have a discernible capability to inhibit tumor formation and the growth of cancer cell, which have a vital link between diet and chronic disease prevention. Several plants and spices that contain flavonoid derivatives have been used in traditional medicine as disease preventative and therapeutic agents. Therefore, flavonoids could be used as chemotherapeutic drugs, indicating their potential clinical utility in cancer treatment. The purpose of this research was to discover and produce innovative pharmaceuticals from natural sources by introducing structural changes into flavonoids' backbones and changing their structures to improve biological activity and anticancer effects. In the current study, it was expected that the percent unbound values for the 15 compounds in human plasma would be low, ranging between 0.188 and 0.391. However, all compounds have a safe range and are not toxic to the brain. Compounds 2, 10, and 13 were shown to be permeable to the CNS (log PS > -3), but all other compounds had difficulty penetrating the CNS. Furthermore, all compounds had a low total clearance, ranging from 0.038 to 1.216 ml/min/kg, indicating that these compounds have a long half-life. None of the compounds caused skin sensitization (SS), and only compounds 1, 11, and 12 are expected to be AMES-positive, suggesting that the other compounds are not mutagenic. The result of the study showed based on the Drug-likeness and ADMET studies, only 3 compounds, including 3, 4, and 15, have a good pharmacokinetics propriety, the lowest toxicity, and good binding affinity towards Caspase 3 V266APDB (ID: 5I9B) as potential inhibitor candidates for the HeLa cell line, they have a low total clearance property and no AMES mutagenicity or hERG inhibition properties. These compounds (3,4,15) were examined to act as new cytotoxic drug candidates and would have an interest as starting point for designing compounds against the HeLa cell line.Communicated by Ramaswamy H. Sarma.

BACKGROUND Since 2016, vaccines against enterovirus 71 (EV71) infection have been approved for use in China. Reports to the national passive surveillance system raised concerns about febrile seizures (FS) after EV71 vaccination. Rapid safety assessment of this novel vaccine is a public health priority. The objective was to assess risks of FS following EV71 vaccination in China. METHODS We used data from a Regional Health Information Platform in Ningbo. The exposed population was children aged 6-71 months who received any dose of EV71 vaccine from 1 January 2016 to 31 December 2019. We implemented a multilayered approach to actively monitor FS following EV71 vaccination that included near real-time surveillance using two complementary sequential designs and further signal evaluation performing self-controlled risk interval (SCRI) analyses. RESULTS A total of 330,668 EV71 doses were administered to the study population. During 157 weeks of sequential analyses, no statistically increased risks were detected, when compared with the self-matched control interval or the background risk. Further SCRI analyses confirmed no associations between EV71 vaccination and FS (adjusted incidence rate ratio: 1.04, 95% CI: 0.75 to 1.43). CONCLUSIONS Our results reassured the safety of FS after EV71 vaccination using postlicensure data for the first time.

Alzheimer's disease (AD) is a multifactorial progressive and irreversible neurodegenerative disorder characterized by severe memory impairment and cognitive disability in the middle and old-aged human population. There are no proven drugs for AD treatment and prevention. In Ayurveda, medhya plants are used to prepare Rasayana, and its consumption improves memory and cognition. Nardostachys jatamansi (D.Don) DC is a medhya plant used in traditional medicine to treat neurological disorders, and its unique pyranocoumarins can be a potential drug candidate for AD. Given its traditional claims, this study aims to find the multi-target potential efficacy of the ligands (drug molecules) against the AD from N. jatamansi pyranocoumarins using computational drug discovery techniques. Drug likeliness analysis confirms that pyranocoumarins of N. jatamansi, such as seselin, jatamansinol, jatamansine, jatamansinone, and dihydrojatamansin are probable drug candidates for AD. Molecular docking, molecular dynamic simulations, and Molecular Mechanics/Generalized Born Surface Area (MM-GBSA) analysis confirm that dihydrojatamansin inhibits acetylcholinesterase (AChE), and jatamansinol inhibits butyrylcholinesterase (BuChE), glycogen synthase kinase 3β (GSK3β), and kelch-like ECH-associating protein 1 (Keap1) AD therapeutic targets. Therefore, this study provides potential multi-target inhibitors that would further validate experimental studies, leading to new treatments for AD.Communicated by Ramaswamy H. Sarma.

Purpose Sorghum is an important cereal crop, cultivated for food, fodder and biofuel. Mutation breeding techniques are used to create genetic variability for qualitative and quantitative traits in crop plants. The purpose of this study was to create induced variability and estimate mutagenic effectiveness and efficiency of physical and chemical mutagens in two sorghum cultivars. Materials and methods Gamma rays (100, 200, 300 and 400 Gy, Co60 source, Bhabha Atomic Research Center, Mumbai, India), ethyl methane sulphonate (0.1%, 0.2%, 0.3%, and 0.4% EMS, Sigma-Aldrich, India) and their combinations were used to mutagenize 296B (rainy season) and Parbhani moti (post-rainy) cultivars. Morphological and yield traits were analyzed for enhanced variability in qualitative and quantitative traits across M2 and M3 generations. Results Based on the mutagenic sensitivity, lethal dose at 50% survivability (LD50) for both the genotypes was found to be 269-281 Gy in case of gamma rays and 0.32-0.33% for EMS. Based on reduced germination and survivability, mutagenic sensitivity was dose dependent and genotype independent. High frequency of chlorophyll mutations (albino, xantha, viridis, variegated and chlorine) were linearly correlated with dose in both the genotypes. Among the favorable mutants, dwarf and brown midrib were isolated from Parbhani moti population, which could be used in the cross breeding programs. A combined treatment, 100Gy +0.1% EMS showed high mutagenic effectiveness and efficiency. Enhanced genetic variation for quantitative traits as measured by wide range values and coefficient of variation was attributed to the effect of physical and chemical mutagens. Early flowering, high grain yield (24-49% increase over control) were identified in M2 and validated in M3 generation in both genotypes. Conclusions This study has revealed wide genetic variability and better effectiveness and efficiency of the physical (300 Gy) and chemical mutagens (0.2%) and their combination (200 Gy+ 0.2%) across two sorghum genotypes. Significant correlations identified between quantitative traits will help in better selection in the segregating generations.

Cakile maritima ssp. aegyptiaca (Wild.) Nyman is growing with dimorphic leaf forms (entire or pinnatifid lamina) along the Mediterranean coast of Egypt. The cytotoxic activities of dried shoot systems of the two morphological forms were evaluated by testing and comparing the effects of ethanolic and aqueous extracts on the viability of five human cell lines. GC-MS analysis was performed to identify the bioactive and anticancer compounds present in the most active extracts. MTT assay indicated that both aqueous and ethanolic extracts have selective cytotoxic activities against cancer cell lines with no inhibitory activities against normal Wi38 or Vero cell lines. The underlying mechanism of cytotoxicity involved the induction of G2/M phase arrest in targeted cells MCF-7 and HCT-116 associated with inducing apoptosis in both cell lines, as indicated by Annexin-V assay. Apoptosis investigation in MCF-7 and HCT-116 cells treated with ethanolic extracts, was further investigated through RT-PCR, which exhibited elevation of proapoptotic genes of P53, BAX, Capase-3,6,7,8,9, and downregulation of antiapoptotic gene (BCL-2) upon treatment. The GC-MS analysis of ethanolic extracts of pinnatifid and entire forms revealed the existence of 18 and 13 compounds, respectively, with eleven compounds that were detected in pinnatifid form only and seven compounds were identified exclusively in the entire form. Molecular Docking study revealed that the identified compounds exhibited good binding affinity towards BCL-2 inhibition, and this agreed with the suggested apoptotic mechanism. To the best of authors' knowledge, this is the first scientific evidence underline the variability in the chemical composition associated with variable anticancer activities of dimorphic forms of C. maritima.Communicated by Ramaswamy H. Sarma.

Hypertrophic cardiomyopathy (HCM) is a common cause of sudden cardiac death in athletes. Cardiac Magnetic Resonance (CMR) imaging is considered an excellent tool to differentiate between HCM and athlete's heart. The aim of this systematic review was to highlight the novel CMR-derived parameters with significant discriminative capacity between the two conditions. A systematic search in the MEDLINE, EMBASE and Cochrane Reviews databases was performed. Eligible studies were considered the ones comparing novel CMR-derived parameters on athletes and HCM patients. Therefore, studies that only examined Cine-derived volumetric parameters were excluded. Particular attention was given to binary classification results from multi-variate regression models and ROC curve analyses. Bias assessment was performed with the Quality Assessment on Diagnostic Accuracy Studies. Five (5) studies were included in the systematic review, with a total of 284 athletes and 373 HCM patients. Several novel indices displayed discriminatory potential, such as native T1 mapping and T2 values, LV global longitudinal strain, late gadolinium enhancement and whole-LV fractal dimension. Diffusion tensor imaging enabled quantification of the secondary eigenvalue angle and fractional anisotropy in one study, which also proved capable of reliably detecting HCM in a mixed athlete/patient sample. Several novel CMR-derived parameters, most of which are currently under development, show promising results in discerning between athlete's heart and HCM. Prospective studies examining the discriminatory capacity of all promising modalities side-by-side will yield definitive answers on their relative importance; diagnostic models can incorporate the best performing variables for optimal results.

INTRODUCTION Intrahepatic cholestasis of pregnancy (ICP) is a liver disease that occurs during pregnancy. While ICP has a minimal impact on the mother, it primarily affects the pregnancy outcome of fetus, resulting in spontaneous miscarriage and even the intrauterine death of fetus. AREAS COVERED This review covers current progress in the role of gut microbiota and bile acids in ICP. EXPERT OPINION The causes and pathogenesis of ICP are currently unclear, and the serum bile acid level is the main clinical evidence for ICP diagnosis. The gastrointestinal tract is home to a tremendous number and type of microbes, which play critical roles in the synthesis and metabolism of bile acids. Studies in recent years have shown that the changes in gut microbiota and bile acid metabolic profiles are closely associated with ICP. This review discusses some of the future prospects in this area of research.

We have previously shown that low-intensity ultrasound (LIUS) can modulate mitochondrial complex I activity and the generation of mitochondrial reactive oxygen species (mROS) in PC12 cells. This study investigated the mechanism of LIUS by comparing its effect on mitochondrial dysfunction by three different pathways. LIUS was shown to reverse the effects of rotenone, a Q-site blocker, on the complex I inhibition, mROS generation, and drop of mitochondrial membrane potential (Δψm). In contrast, common antioxidants, N-acetyl cysteine (NAC) and uric acid (UA), blocked rotenone-induced mROS generation and Δψm drop without recovering the complex I activity, which suggested that Δψm drop is correlated with mROS generation rather than complex I inhibition itself. Ionomycin, an ionophore for Ca2+, and L-buthionine-S,R-sulfoximine (BSO), an inhibitor of glutathione (GSH) biosynthesis, induced mROS generation and Δψm drop without inhibiting complex I activity via different mechanisms. LIUS showed no effect on ionomycin-induced Δψm drop but showed partial inhibition on the other effects of ionomycin and BSO. These results suggest that LIUS might have redundant mechanisms but acted mainly on the complex I activity thereby modulating mROS and Δψm levels. LIUS appeared to act on the Q-module of complex I because it showed no inhibitory effect on Zn2+, an inhibitor of the proton transporting P-module of complex I. Interestingly, pre-treatment of LIUS for up to an hour in advance blocked the rotenone effect as efficiently as the co-treatment. Further studies are needed to reveal the exact mechanism of LIUS to inhibit complex I activity.

Microbial-assisted removal of natural or synthetic pollutants is the prevailing green, low-cost technology to treat polluted environments. However, the challenge with enzyme-assisted bioremediation is the laborious nature of dehalogenase-producing microorganisms’ bioprospecting. This bottleneck could be circumvented by in-silico analysis of certain microorganisms’ whole-genome sequences to predict their protein functions and enzyme versatility for improved biotechnological applications. Herein, this study performed structural analysis on a dehalogenase (DehHsAAD6) from the genome of Halomonas smyrnensis AAD6 by molecular docking and molecular dynamic (MD) simulations. Other bioinformatics tools were also employed to identify substrate preference (haloacids and haloacetates) of the DehHsAAD6. The DehHsAAD6 preferentially degraded haloacids and haloacetates (−3.2–4.8 kcal/mol) and which formed three hydrogen bonds with Tyr12, Lys46, and Asp182. MD simulations data revealed the higher stability of DehHsAAD6-haloacid- (RMSD 0.22–0.3 nm) and DehHsAAD6-haloacetates (RMSF 0.05–0.14 nm) complexes, with the DehHsAAD6-L-2CP complex being the most stable. The detail of molecular docking calculations ranked complexes with the lowest binding free energies as: DehHsAAD6-L-2CP complex (−4.8 kcal/mol) = DehHsAAD6-MCA (−4.8 kcal/mol) < DehHsAAD6-TCA (−4.5 kcal/mol) < DehHsAAD6-2,3-DCP (−4.1 kcal/mol) < DehHsAAD6-D-2CP (−3.9 kcal/mol) < DehHsAAD6-2,2-DCP (−3.5 kcal/mol) < DehHsAAD6-3CP (−3.2 kcal/mol). In a nutshell, the study findings offer valuable perceptions into the elucidation of possible reaction mechanisms of dehalogenases for extended substrate specificity and higher catalytic activity. Communicated by Ramaswamy H. Sarma

Purpose Inflammatory bowel diseases (IBDs) are global health problems that are associated with immune regulation, but clinical IBDs treatment is currently inadequate. Effective preventive or therapeutic methods for immune disorders rely on controlling the function of immune cells. Isosteviol sodium (STV-Na) has antioxidant activity, but the therapeutic effect of STV-Na against IBD remain undocumented. Herein, we investigated the therapeutic effect of STV-Na in mice models with IBDs. Methods Mice received 3.5% DSS for 7 days to establish IBD models. Intraperitoneal STV-Na was given 2 days before DSS and lasted for 9 days. Commercially available drugs used in treating IBDs (5-aminosalicylic acid, dexamethasone, and infliximab) were used as positive controls. Samples were collected 7 days after colitis induction. Histopathological score, biochemical parameters, molecular biology methods, and metabolomics were used for evaluating the therapeutic effect of STV-Na. Results Our data revealed that STV-Na could significantly alleviate colon inflammation in mice with colitis. Specifically, STV-Na treatment improved body weight loss, increased colon length, decreased histology scores, and restored the hematological parameters of mice with colitis. The untargeted metabolomics analysis revealed that metabolic profiles were restored by STV-Na treatment. Furthermore, STV-Na therapy suppressed the number of CD68 macrophages and F4/80 cell infiltration. And STV-Na suppressed M1 and M2 macrophage numbers along with the mRNA expressions of proinflammatory cytokines. Moreover, STV-Na administration increased the number of regulatory T (Treg) cells while decreasing Th1/Th2/Th17 cell counts in the spleen. Additionally, STV-Na treatment restored intestinal barrier disruption in DSS-triggered colitis tissues by ameliorating the TJ proteins, increasing goblet cell proportions, and mucin protein production, and decreasing the permeability to FITC-dextran, which was accompanied by decreased plasma LPS and DAO contents. Conclusion These results indicate that STV-Na can ameliorate colitis by modulating immune responses along with metabolic reprogramming, and could therefore be a promising therapeutic strategy for IBDs., Graphical Abstract

PrEP use has steadily increased since its introduction, yet uptake remains slow among MSM of color in the U.S. South. Findings presented are from a qualitative study conducted in 2016 exploring factors related to remaining HIV negative among Black (n = 99) and Latino (n = 51) MSM in Atlanta, Baton Rouge, New Orleans, Jackson, and Miami. One-hour in-depth interviews were analyzed using a qualitative content analysis approach. MSM perceived PrEP as providing relief from fear and as an insurance policy against HIV infection but were likely to consider themselves at low risk for HIV. Identified factors influencing PrEP use included side effects, medical mistrust, and stigma. Cost and access were not seen as major barriers. Findings suggest MSM of color in the South may view PrEP as too risky for their HIV prevention needs. PrEP providers can address medical mistrust, discuss side effects, and emphasize positive aspects of PrEP use to increase uptake. (150 words).

The Asia-Pacific region is renowned to harbor nearly half of the global hot spots of biodiversity. Accordingly, many endemic species of boletes have already been recorded from this geographic region. However, the majority of the specific descriptions of reported boletoid species follow classical concepts of taxonomy, and by comparison only a few taxa have been corroborated by modern molecular techniques. In this study, we focused on specimens in a new clade uncovered by our previous studies. By careful reexamination of macroscopic and microscopic characters of Boletus granulopunctatus, originally described from Japan, and Xerocomus mcrobbii, originally described from New Zealand, we discovered a new genus and species Amoenoboletus miraculosus from Sabah, Malaysia. In addition, three new combinations in Amoenoboletus are proposed, and a dichotomous key to species in the genus is provided. The phylogenetically close relationship among Amoenoboletus species suggests a tight geographic correlation in the Asia-Pacific region.

A novel beer-spoilage bacterium, Lactobacillus acetotolerans, has frequently been encountered in south China beer breweries, imparting an undesirable and unpalatable “buttery” taste and oily mouthf...