Pathogenic Changes Research Articles

Genetic Etiology Investigation in Treatment-Resistant Nocturnal Enuresis Children: A descriptive study.

Our study aimed to evaluate the genetic etiology of treatment-resistant nocturnal enuresis in children who have undergone at least 6 episodes of behavioral therapy, urotherapy, alarm therapy, and medical treatment. A total of 21 patients were included in the study. Inclusion criteria for the study comprised children aged 5-18 years diagnosed with treatment-resistant enuresis according to the International Children's Continence Society (ICCS) guidelines. The capture-based Sophia Hereditary Disease Panel by Sophia Genetics was used specifically for nocturnal enuresis, consisting of a panel of 19 genes (AGXT, AQP2, AVPR2, BNC2, CLCNKB, DLG3, ELN, FA2H, FAM20A, FOXP1, HPSE2, KCNJ10, MLXIPL, NPHP3, RNF168, SLC12A3, SLC25A13, SLC5A2, SMARCA2). Patients were analyzed for genetic variations in genes associated with nocturnal enuresis, including AGXT, AQP2, AVPR2, BNC2, CLCNKB, DLG3, ELN, FA2H, FAM20A, FOXP1, HPSE2, KCNJ10, MLXIPL, NPHP3, RNF168, SLC12A3, SLC25A13, SLC5A2, and SMARCA2. No pathogenic changes potentially explaining the etiology of the disease were detected in 20 patients. One patient exhibited a variant in the AQP2 gene at hg19:Chr12:50344908 exon 1, c.295G>A locus, classified as a Variant of Uncertain Significance (VUS) according to the American College of Medical Genetic and Genomics (ACMG) 2015 guidelines. The AQP2 gene is associated with autosomal dominant and autosomal recessive inherited nephrogenic diabetes insipidus (type 2) in the OMIM (Online Mendelian Inheritance in Man) database. Our study resembles studies indicating that nocturnal enuresis cases do not have a monogenic etiology but occur with multifactorial effects and have a weak correlation between genotype and phenotype.

Post-penetration development, pathogenicity and histological changes in ridge gourd infected by root-knot nematode (Meloidogyne incognita)

Post-penetration development, pathogenicity and histological changes in ridge gourd infected by root-knot nematode (Meloidogyne incognita)

Skeletal myotubes expressing ALS mutant SOD1 induce pathogenic changes, impair mitochondrial axonal transport, and trigger motoneuron death

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the loss of motoneurons (MNs), and despite progress, there is no effective treatment. A large body of evidence shows that astrocytes expressing ALS-linked mutant proteins cause non-cell autonomous toxicity of MNs. Although MNs innervate muscle fibers and ALS is characterized by the early disruption of the neuromuscular junction (NMJ) and axon degeneration, there are controversies about whether muscle contributes to non-cell-autonomous toxicity to MNs. In this study, we generated primary skeletal myotubes from myoblasts derived from ALS mice expressing human mutant SOD1G93A (termed hereafter mutSOD1). Characterization revealed that mutSOD1 skeletal myotubes display intrinsic phenotypic and functional differences compared to control myotubes generated from non-transgenic (NTg) littermates. Next, we analyzed whether ALS myotubes exert non-cell-autonomous toxicity to MNs. We report that conditioned media from mutSOD1 myotubes (mutSOD1-MCM), but not from control myotubes (NTg-MCM), induced robust death of primary MNs in mixed spinal cord cultures and compartmentalized microfluidic chambers. Our study further revealed that applying mutSOD1-MCM to the MN axonal side in microfluidic devices rapidly reduces mitochondrial axonal transport while increasing Ca2 + transients and reactive oxygen species (i.e., H2O2). These results indicate that soluble factor(s) released by mutSOD1 myotubes cause MN axonopathy that leads to lethal pathogenic changes.

Identification and comparison of protein composition of biofilms in response to EGCG from Enterococcus faecalis and Staphylococcus lugdunensis, which showed opposite patterns in biofilm-forming abilities

Bacterial biofilm is resistant to conventional antibiotic treatments, leading to complications associated with many infection-related human diseases. Epigallocatechin Gallate (EGCG), a phenolic catechin enriched in green tea, is recognized for its anti-bacterial and anti-biofilm activities. In this study, we examined the protein components of the biofilms formed in the absence or presence of EGCG using Enterococcus faecalis and Staphylococcus lugdunensis, which had shown opposing patterns in biofilm formation. A clustering heatmap revealed that the two microorganisms expressed the different protein sets in response to EGCG. Proteins that were noticeably upregulated included those associated with stress responsiveness and gluconeogenesis in E. faecalis, and gene modification in S. lugdunensis. Conversely, downregulated proteins were related to tRNA-modifying enzyme activity in E. faecalis, and anabolic metabolism in S. lugdunensis. Among the proteins identified only in EGCG-responsive biofilms, enzymes involved in de novo purine biosynthesis were enriched in E. faecalis, while proteins likely to cause DNA instability and pathogenicity changes were abundantly present in S. lugdunensis. The classification based on gene ontology (GO) terms by microorganism exhibited that metabolic process or catabolic activity was at the top rank in E. faecalis with more than 33 proteins, and in S. lugdunensis, localization or transport was highly ranked with 4 proteins. These results support the hypothesis that EGCG might cause different cellular programs in each microorganism. Finally, comparison of the proteomes between two groups that form biofilms to similar extents discovered that 2 proteins were commonly found in the weak biofilm-forming groups (E. faecalis and EGCG-responding S. lugudunensis), whereas 9 proteins were common among the strong biofilm-forming groups (S. lugdunensis and EGCG-responding E. faecalis). It was suggested that these proteins could serve as potential indicators to detect the presence and predict the extent of biofilm formation by multiple microorganisms. Taken all together, proteomics data and analyses performed in this study provided useful and new information on the proteins embedded in the biofilms formed at the specific conditions, which can aid in diagnosis and the development of tailored treatment strategies.

Increased pathogen exposure of a marine apex predator over three decades.

Environmental changes associated with global warming create new opportunities for pathogen and parasite transmission in Arctic wildlife. As an apex predator ranging over large, remote areas, changes in pathogens and parasites in polar bears are a useful indicator of changing transmission dynamics in Arctic ecosystems. We examined prevalence and risk factors associated with exposure to parasites and viral and bacterial pathogens in Chukchi Sea polar bears. Serum antibodies to six pathogens were detected and prevalence increased between 1987-1994 and 2008-2017 for five: Toxoplasma gondii, Neospora caninum, Francisella tularensis, Brucella abortus/suis, and canine distemper virus. Although bears have increased summer land use, this behavior was not associated with increased exposure. Higher prevalence of F. tularensis, Coxiella burnetii, and B. abortus/suis antibodies in females compared to males, however, could be associated with terrestrial denning. Exposure was related to diet for several pathogens indicating increased exposure in the food web. Elevated white blood cell counts suggest a possible immune response to some pathogens. Given that polar bears face multiple stressors in association with climate change and are a subsistence food, further work is warranted to screen for signs of disease.

Exploring the Phenotypic Spectrum of KCNB1: A Case Series of Two Unique Presentations

Background and Purpose: We present two cases exemplifying the broad phenotypic spectrum associated with pathogenic changes in KCNB1 protein. Our case series highlights the variability of the outcomes of this genetic condition. Methods: This is a descriptive retrospective review of medical histories of two patients under the age of 18 years with KCNB1 related neurodevelopmental disorder. We performed a literature review from published articles in PubMed indexed journals. Brief Report: A 9-year-old girl with developmental delay and autism found to have a pathogenic mutation in the KCNB1 gene presented with regression in her language, staring spells, and EEG findings suggestive of Electrographic Status Epilepticus in Sleep (ESES). She is maintained on Clobazam and continues to make gains in her milestones. A 16-year-old boy with a heterozygous mutation in the KCNB1 gene presented with mild intellectual disability and attention deficit hyperactivity disorder. His EEG showed frequent bursts of generalized, sharply contoured theta activity - a finding of uncertain clinical significance. However, no clinical seizures were mentioned. He is maintained on valproic acid for mood stabilization. Conclusion: The phenotype associated with KCNB1-related neurodevelopmental disorders is variable and includes epilepsy, intellectual disabilities, and psychiatric diagnoses. While our first patient had typical manifestations of the condition, treatment with anti-seizure medication was seen as beneficial. Our second patient had no clinical seizures, and his case highlights the importance of assessing the risk-benefit ratio of anti-seizure medication. It is important to understand the spectrum of the disease in order to identify clinical manifestations and perform early testing for a better prognosis.

Modern Didactics of Acute Pneumonia as a Side Effect of the Antibiotic Era

The widespread and uncontrolled use of antibiotics for more than eighty years, including not only medicine but also the food industry, has had a significant impact on the usual relationships between representatives of the microbiosphere that accompanies our body. For a long time, the possibility of side effects remained without due attention, giving way to attempts to maintain the original antimicrobial effect of these drugs. Currently, evidence of the consequences of antibiotic therapy has received official recognition only in the form of resistant microflora. Phenomena such as the constant change of AP pathogens and the gradual loss of antibiotics for their purpose remain unstudied. The selective nature of specialists' attention to the side effects of antibiotics is due to a decrease in their effectiveness and the desire to restore the successes of previous therapy. The latter circumstance is a consequence of the negative didactic influence of antibiotics on professional views that determine the strategy for solving the problem and require, first of all, changes following the fundamental canons of medical science and numerous facts.

Inflammatory profile of eosinophils in asthma-COPD overlap and eosinophilic COPD: a multi-omics study.

Elevated blood eosinophil levels in patients with chronic obstructive pulmonary disease (COPD) with or without asthma are linked to increased exacerbations and the effectiveness of inhaled corticosteroid treatment. This study aimed to delineate the inflammatory cellular properties of eosinophils in patients with asthma-COPD overlap (ACO) and eosinophilic COPD (eCOPD). Eosinophils were isolated from the peripheral blood of healthy volunteers, patients with non-eCOPD, and those with ACO/eCOPD. Multi-omics analysis involving transcriptomics, proteomics, and lipidomics was performed, followed by bioinformatic data analyses. In vitro experiments using eosinophils from healthy volunteers were conducted to investigate the molecular mechanisms underlying cellular alterations in eosinophils. Proteomics and transcriptomics analyses revealed cellular characteristics in overall COPD patients represented by viral infection (elevated expression of sterol regulatory element-binding protein-1) and inflammatory responses (elevated levels of IL1 receptor-like 1, Fc epsilon receptor Ig, and transmembrane protein 176B). Cholesterol metabolism enzymes were identified as ACO/eCOPD-related factors. Gene Ontology and pathway enrichment analyses demonstrated the key roles of antiviral responses, cholesterol metabolism, and inflammatory molecules-related signaling pathways in ACO/eCOPD. Lipidomics showed the impaired synthesis of cyclooxygenase-derived mediators including prostaglandin E2 (PGE2) in ACO/eCOPD. In vitro assessment confirmed that IL-33 or TNF-α stimulation combined with IL-5 and IFN-γ stimulation induced cellular signatures in eosinophils in ACO/eCOPD. Atorvastatin, dexamethasone, and PGE2 differentially modulated these inflammatory changes. ACO/eCOPD is associated with viral infection and an inflammatory milieu. Therapeutic strategies using statins and inhaled corticosteroids are recommended to control these pathogenic changes.

Inactivation induced by pathogenic Cav1.3 L-type Ca2+-channel variants enhances sensitivity for dihydropyridine Ca2+ channel blockers.

Pathogenic gain-of-function mutations in Cav1.3 L-type voltage-gated Ca2+-channels (CACNA1D) cause neurodevelopmental disorders with or without endocrine symptoms. We aimed to confirm a pathogenic gain-of function phenotype of CACNA1D de novo missense mutations A749T and L271H, and investigated the molecular mechanism causing their enhanced sensitivity for the Ca2+-channel blocker isradipine, a potential therapeutic for affected patients. Wildtype and mutant channels were expressed in tsA-201 cells and their gating analysed using whole-cell and single-channel patch-clamp recordings. The voltage-dependence of isradipine action was quantified using protocols inducing variable fractions of inactivated channels. The molecular basis for altered channel gating in the mutants was investigated using in silico modelling and molecular dynamics simulations. Both mutations were confirmed pathogenic due to characteristic shifts of voltage-dependent activation and inactivation towards negative potentials (~20 mV). At negative holding potentials both mutations showed significantly higher isradipine sensitivity compared to wildtype. The affinity for wildtype and mutant channels increased with channel inactivation as predicted by the modulated receptor hypothesis (30- to 40-fold). The IC50 was indistinguishable for wildtype and mutants when >50% of channels were inactivated. Mutations A749T and L271H induce pathogenic gating changes. Like wildtype, isradipine inhibition is strongly voltage-dependent. Our data explains their apparent higher drug sensitivity at a given negative voltage by the availability of more inactivated channels due to their more negative inactivation voltage range. Low nanomolar isradipine concentrations will only inhibit Cav1.3 channels in neurons during prolonged depolarized states without selectivity for mutant channels.

Intraspecific cooperation allows the survival of Staphylococcus aureus staff: a novel strategy for disease relapse

BackgroundThe contribution of interspecies interactions between coinfecting pathogens to chronic refractory infection by affecting pathogenicity is well established. However, little is known about the impact of intraspecific interactions on infection relapse, despite the cross-talk of different strains within one species is more common in clinical infection. We reported a case of chronic refractory pulmonary infection relapse, caused by two methicillin-sensitive S. aureus (MSSA) strains (SA01 and SA02) and revealed a novel strategy for relapse via intraspecific cooperation.MethodsThe hemolytic ability, growth curve, biofilm formation, virulence genes and response of G. mellonella larvae to S. aureus infection were analysed to confirm this hypothesis.ResultsSA02 hemolytic activity was inhibited by SA01, along with the expression of hemolysin genes and the virulence factor Hla. Additionally, SA01 significantly enhanced the biofilm formation of SA02. AIP-RNAIII may be a possible pathway for this interaction. Compared with mono-infection, a worse outcome (decreased larval survival and increased microbial burden) of the two MSSA strains coinfected with G. mellonella confirmed that intraspecific interactions indeed enhanced bacterial survival in vivo.ConclusionThe intraspecific interaction of S. aureus could lead to chronic refractory infection via pathogenicity changes.

Chronic Peroxisome Proliferator-Activated Receptor α/γ and Cannabinoid Receptor 2 Agonist Treatments Attenuated Visceral Adipose Tissue (VAT)–Derived Extracellular Vesicle–Related VAT and Intestinal Abnormalities in Nonalcoholic Steatohepatitis Mice

This study explores the mechanisms and combined effects of chronic peroxisome proliferator-activated receptor (PPAR)α/γ and cannabinoid receptor 2 (CB2R) agonists on visceral adipose tissue (VAT)-derived extracellular vesicle (EVs) release and associated systemic/VAT inflammation, decreased VAT capillary density/fibrosis, and intestinal inflammation/hyperpermeability in nonalcoholic steatohepatitis (NASH) mice. NASH mice receiving 1 month of PPARα/γ agonist aleglitazar (10 mg/kg/day), CB2R agonist JWH015 (3 mg/kg/day) alone or combined. High EV release from VAT of NASH mice was associated with severe systemic/VAT/intestinal inflammation, reduced capillary network of VAT, and intestinal hyperpermeability. Combined JWH015 with aleglitazar treatment significantly suppressed HFD-induced obesity/adiposity, inhibited VAT expansion, reduced VAT inflammation/fibrosis, normalized VAT capillary network, and attenuated intestinal mucosal injury, inflammation, and hyperpermeability in NASH+aleg+JWH015 mice. The inhibition of AT-derived EV release and hypoxia inducible factor (HIF)1α levels in AT-derived EV, normalization of CB2R, PPARα, PPARγ, PPARγ1, PPARγ2, tight junction proteins, VEGF/CD31 expression, and downregulations of HIF1α, MCP-1 and TGFβ1 were observed in the VAT and intestine of the NASH+aleg+jwh015 group. In vitro experiments revealed that PPARα/γ and CB2R activation attenuated NASH AT-derived EV (EVnash)-induced pathogenic changes in the J774/SVEC4-10/Caco2 /3T3-L1 cell system. This study suggested that VAT-derived EVs contribute to the pathogenesis of NAFLD and that combined PPARα/γ and CB2R agonist treatment reduces VAT-released EV release and HIF1/MCP-1 signals to ameliorate hepatic steatosis and VAT/intestine abnormalities of NASH mice.

Trends in surveillance indicators for central-catheter-associated bloodstream infections in a tertiary hospital in Switzerland

Central line-associated bloodstream infections (CLABSIs) are associated with relevant morbidity and mortality. Longitudinal dynamics in CLABSI incidence and the spectrum of causative microorganisms are limited. To describe trends in CLABSI incidence, use of central lines, and causative pathogens. We analysed prospectively collected data on CLABSI at a Swiss tertiary care hospital between January 2016 and December 2023. We investigated longitudinal changes of incidence densities, catheter utilization and causative pathogens. A total of 707 CLABSIs were observed, corresponding to an incidence density of 1.69 (95% confidence interval 1.56-1.81) CLABSIs per 1000 catheter-days. There was no significant trend of CLABSI incidence density per 1000 catheter-days (z= 0.86, P=0.391), but an increase of catheter utilization ratio (z= 8.88, P<0.001). Coagulase-negative staphylococci (N= 207, 23.4%) and Enterococcus spp. (N= 134, 15.2%; E.faecium N= 94, 10.6%; E.faecalis N= 37, 4.1%; other Enterococcus spp. N= 3, 0.34%) were the most frequent causative pathogens. Over the years, the proportion of Enterococcus spp. (z= 3.4, P<0.001), driven by an increase of E.faecium (z= 3.2, P=0.001), and yeast (z= 2.3, P=0.020) increased, whereas coagulase-negative staphylococci decreased (z= -6.1, P<0.001). Prospective CLABSI surveillance indicated stable incidence densities per 1000 catheter-days, but there were significant shifts of causative microorganisms over time.

Sexual behavior is linked to changes in gut microbiome and systemic inflammation that lead to HIV-1 infection in men who have sex with men

Pathogenic changes in gut microbial composition precede the onset of HIV-1 infection in men who have sex with men (MSM). This process is associated with increased levels of systemic inflammatory biomarkers and risk for AIDS development. Using mediation analysis framework, in this report we link the effects of unprotected receptive intercourse among MSM prior to primary HIV-1 infection to higher levels of proinflammatory cytokines sCD14 and sCD163 in plasma and a significant decrease in the abundance of A. muciniphila, B. caccae, B. fragilis, B. uniformis, Bacteroides spp., Butyricimonas spp., and Odoribacter spp., and a potential increase in the abundance of Dehalobacterium spp. and Methanobrevibacter spp. in stools of MSM with the highest number of sexual partners. These differences in microbiota, together with a reduction in the pairwise correlations among commensal and short-chain fatty acid-producing bacteria with a number of sexual partners, support an increase in gut dysbiosis with the number of sexual partners. These results demonstrate the interconnectedness of sexual behavior, immune response, and microbiota composition, notably among MSM participating in high-risk sexual behaviors.

Surge in Mycoplasma Pneumoniae infection and Respiratory Viruses Co-infection in Children With Community-Acquired Pneumonia in the Post-Pandemic.

During the COVID-19 pandemic, multifaceted non-pharmaceutical interventions have not only reduced the transmission of SARS-CoV2 but also affected the prevalence of other respiratory pathogens. With the lifting of many restrictions, a surge in cases of pneumonia in children has been reported in many hospitals in China. The study assessed the changes in pathogen and symptoms of children with community-acquired pneumonia (CAP) before and after the adjustments of prevention and control measures of epidemic and provided recommendations for CAP in children. Children diagnosed with CAP were enrolled in the study from 2022 to 2023. A cross-sectional retrospective study was conducted in a general hospital. We analyzed the data about demographic data, clinical symptoms, pathogens, and medical treatments. The Chi-square and Mann-Whitney U-test were used to assess the statistical significance of groups. We studied 1103 children, 339 in 2022 and 764 in 2023. Compared with children in 2022, more children were diagnosed with CAP in 2023 and these children had a higher body temperature and levels of CRP and PCT, which indicated these children got severe inflammation. The positive rate of the pathogen was also higher in 2023, especially the detective rate of Mycoplasma pneumoniae. The number of children infected with more than two pathogens was higher in 2023, especially those co-infected with the virus and M. Pneumoniae. Concerning the medicine therapy, the usage of β-lactam antibiotics, Macrolide antibiotics, and antiviral drugs kept rapid growth. After the adjustment of epidemic prevention and control policies in 2023, more children got CAP with severe clinical symptoms, and more antibiotics and antiviral drugs were used. Further study is needed to explore the reasons for the increase in children with CAP and to explore the rationality of treatment.

Genome characterization of Rift Valley fever virus isolated from cattle, goats and sheep during interepidemic periods in Kenya

Rift Valley fever virus (RVFV) is a mosquito-borne RNA virus of the Phlebovirus genus in the phenuviridae family. Its genome is trisegmented with small (S), medium (M) and large (L) fragments. In nature, the virus exists as a single serotype that is responsible for outbreaks of Rift Valley fever (RVF), a zoonotic disease that often occurs in Africa and the Middle East. RVFV genomes are thought to undergo both recombination and reassortment and investigations of these events is important for monitoring the emergence of virulent strains and understanding the evolutionary characteristics of this virus. The aim of this study was to characterize the genomes of RVFV isolates from cattle, sheep, and goats collected during an interepidemic period in Kenya between June 2016 and November 2021. A total of 691 serum samples from cattle (n = 144), goats (n = 185) and sheep (n = 362) were analysed at the Central Veterinary Laboratories. The competitive IgM-capture ELISA, was used to screen the samples; 205 samples (29.67%) tested positive for RVFV. Of the 205 positive samples, 42 (20.5%) were from cattle, 57 (27.8%) from goats, and 106 (51.7%) from sheep. All the IgM-positive samples were further analyzed by qPCR, and 24 (11.71%) tested positive with Ct values ranging from 14.788 to 38.286. Two samples, 201808HABDVS from sheep and 201810CML3DVS from cattle, had Ct values of less than 20.0 and yielded whole genome sequences with 96.8 and 96.4 coverage, respectively. There was no statistically significant evidence of recombination in any of the three segments and also phylogenetic analysis showed no evidence of reassortment in the two isolated RVFV segments when compared with other isolates of different lineages from previous outbreaks whose genomes are deposited in the GenBank. No evidence of reassortment leaves room for other factors to be the most probable contributors of change in virulence, pathogenicity and emergence of highly virulent strains of the RVFV.

Stabilizing microtubules aids neurite structure and disrupts syncytia formation in human cytomegalovirus-infected human forebrain neurons.

Human cytomegalovirus (HCMV) is a prolific human herpesvirus that infects most individuals by adulthood. While typically asymptomatic in adults, congenital infection can induce serious neurological symptoms including hearing loss, visual deficits, cognitive impairment, and microcephaly in 10-15% of cases. HCMV has been shown to infect most neural cells with our group recently demonstrating this capacity in stem cell-derived forebrain neurons. Infection of neurons induces deleterious effects on calcium dynamics and electrophysiological function paired with gross restructuring of neuronal morphology. Here, we utilize an iPSC-derived model of the human forebrain to demonstrate how HCMV infection induces syncytia, drives neurite retraction, and remodels microtubule networks to promote viral production and release. We establish that HCMV downregulates microtubule associated proteins at 14 days postinfection while simultaneously sparing other cytoskeletal elements, and this includes HCMV-driven alterations to microtubule stability. Further, we pharmacologically modulate microtubule dynamics using paclitaxel (stabilize) and colchicine (destabilize) to examine the effects on neurite structure, syncytial morphology, assembly compartment formation, and viral release. With paclitaxel, we found improvement of neurite outgrowth with a corresponding disruption to HCMV-induced syncytia formation and Golgi network disruptions but with limited impact on viral titers. Together, these data suggest that HCMV infection-induced disruption of microtubules in human cortical neurons can be partially mitigated with microtubule stabilization, suggesting a potential avenue for future neuroprotective therapeutic exploration.

GENOMIC DIVERSITY OF TOXIGENIC VIBRIO CHOLERAE O1 BIOVAR EL TOR STRAINS ISOLATED DURING THREE WAVES OF THE 7TH CHOLERA PANDEMIC

Introduction. High genome variability of the 7th cholera pandemic agent, V. cholerae El Tor, led to emergence of genovariants with distinct set of altered genes. The aim of the work was to analyze the dynamics of changes in pathogenicity, epidemicity as well as drug resistance and phylogeny in toxigenic strains of V. cholerae El Tor isolated in Russia and endemic regions during three waves of ongoing pandemic. Materials and methods. We used whole-genome nucleotide sequences of 155 strains, obtained by us (42) and taken from the NCBI Genbank (113). DNA sequencing was performed on Ion PGM platform. Phylogenetic relations were determined based on the Bayesian analysis of core SNPs obtained using Snippy 4.6 software package. Antibiotic resistance was assessed applying disk diffusion test. Results. SNP data revealed that the studied strains (1970-2023) might be divided into three clusters. A clear correlation between each-cluster strain genotype and relevant isolation timing was observed. Separation of genetically altered cluster II and III strains isolated during the 2nd and 3rd waves of the pandemic from typical cluster I strains is associated with acquisition of new DNA regions and mutations in pathogenicity and drug resistance genes. Due to different combination of mutations, cluster III strains are genetically heterogeneous. Genome comparison showed that this diversity increased dramatically during the 3rd wave, which led to emergence of new genovariants with higher pathogenic and epidemic potential. It is demonstrated that antibiotic resistance in strains both from endemic regions and Russia over the past 30 years (1993-2023) has undergone significant changes. Thereat, the changing drug resistance clearly correlated with the occurrence of mutations in various pathogenicity genes. Conclusion. It is shown that over the past two decades the agent genome underwent a rather rapid change resulting in emergence of various genovariants. A change in the pathogen variants in Russia has been established. Strains combining genetic markers of hyper-virulence and multiple drug resistance are of particular concern. Genome variability of the strains identified in points at a need for constant genomic surveillance to obtain data on altering epidemically important properties for timely generation of new diagnostic and preventive means.

Pathogenicity of Duck Adenovirus Type 3 in Chickens.

Duck adenovirus Type 3 (DAdV-3) severely affects the health of ducks; however, its pathogenicity in chickens remains unknown. The objectives of this study were to evaluate the pathogenicity and major pathological changes caused by DAdV-3 in chickens. Viral DNA was extracted from the liver of the Muscovy duck, and the fiber-2 and hexon fragments of DAdV-3 were amplified through polymerase chain reaction (PCR). The evolutionary tree revealed that the isolated virus belonged to DAdV-3, and it was named HE-AN-2022. The mortality rate of chicks that received inoculation with DAdV-3 subcutaneously via the neck was 100%, while the mortality rate for eye-nose drop inoculation was correlated with the numbers of infection, with 26.7% of chicks dying as a result of exposure to multiple infections. The main symptoms exhibited prior to death were hepatitis-hydropericardium syndrome (HHS), ulceration of the glandular stomach, and a swollen bursa with petechial hemorrhages. A histopathological examination revealed swelling, necrosis, lymphocyte infiltration, and basophilic inclusion bodies in multiple organs. Meanwhile, the results of quantitative real-time PCR (qPCR) demonstrated that DAdV-3 could affect most of the organs in chickens, with the gizzard, glandular stomach, bursa, spleen, and liver being the most susceptible to infection. The surviving chicks had extremely high antibody levels. After the chickens were infected with DAdV-3 derived from Muscovy ducks, no amino acid mutation was observed in the major mutation regions of the virus, which were ORF19B, ORF66, and ORF67. On the basis of our findings, we concluded that DAdV-3 infection is possible in chickens, and that it causes classic HHS with ulceration of the glandular stomach and a swollen bursa with petechial hemorrhages, leading to high mortality in chickens. The major variation domains did not change in Muscovy ducks or in chickens after infection. This is the first study to report DAdV-3 in chickens, providing a new basis for preventing and controlling this virus.

Strain variation and anomalous climate synergistically influence cholera pandemics.

Explanations for the genesis and propagation of cholera pandemics since 1817 have remained elusive. Evolutionary pathogen change is presumed to have been a dominant factor behind the 7th "El Tor" pandemic, but little is known to support this hypothesis for preceding pandemics. The role of anomalous climate in facilitating strain replacements has never been assessed. The question is of relevance to guide the understanding of infectious disease emergence today and in the context of climate change. We investigate the roles of climate and putative strain variation for the 6th cholera pandemic (1899-1923) using newly assembled historical records for climate variables and cholera deaths in provinces of former British India. We compare this historical pandemic with the 7th (El Tor) one and with the temporary emergence of the O139 strain in Bangladesh and globally. With statistical methods for nonlinear time series analysis, we examine the regional synchrony of outbreaks and associations of the disease with regional temperature and rainfall, and with the El Niño Southern Oscillation (ENSO). To establish future expectations and evaluate climate anomalies accompanying historical strain replacements, climate projections are generated with multi-model climate simulations for different 50-year periods. The 6th cholera pandemic featured the striking synchronisation of cholera outbreaks over Bengal during the El Niño event of 1904-07, following the invasion of the Bombay Presidency with a delay of a few years. Accompanying anomalous weather conditions are similar to those related to ENSO during strain replacements and pandemic expansions into Africa and South America in the late 20th century. Rainfall anomalies of 1904-05 at the beginning of the large cholera anomaly fall in the 99th percentile of simulated changes for the regional climate. Evolutionary pathogen change can act synergistically with climatic conditions in the emergence and propagation of cholera strains. Increased climate variability and extremes under global warming provide windows of opportunity for emerging pathogens.

Metabolic Insights into Neuropsychiatric Illnesses and Ketogenic Therapies: A Transcriptomic View.

The disruption of brain energy metabolism, leading to alterations in synaptic signaling, neural circuitry, and neuroplasticity, has been implicated in severe mental illnesses such as schizophrenia, bipolar disorder, and major depressive disorder. The therapeutic potential of ketogenic interventions in these disorders suggests a link between metabolic disturbances and disease pathology; however, the precise mechanisms underlying these metabolic disturbances, and the therapeutic effects of metabolic ketogenic therapy, remain poorly understood. In this study, we conducted an in silico analysis of transcriptomic data to investigate perturbations in metabolic pathways in the brain across severe mental illnesses via gene expression profiling. We also examined dysregulation of the same pathways in rodent or cell culture models of ketosis, comparing these expression profiles to those observed in the disease states. Our analysis revealed significant perturbations across all metabolic pathways, with the greatest perturbations in glycolysis, the tricarboxylic acid (TCA) cycle, and the electron transport chain (ETC) across all three disorders. Additionally, we observed some discordant gene expression patterns between disease states and ketogenic intervention studies, suggesting a potential role for ketone bodies in modulating pathogenic metabolic changes. Our findings highlight the importance of understanding metabolic dysregulation in severe mental illnesses and the potential therapeutic benefits of ketogenic interventions in restoring metabolic homeostasis. This study provides insights into the complex relationship between metabolism and neuropsychiatric disorders and lays the foundation for further experimental investigations aimed at appreciating the implications of the present transcriptomic findings as well as developing targeted therapeutic strategies.