Pathogenic Markers Research Articles

Existence of Thermotolerant and Salt-Loving Diarrheagenic Vibrio alginolyticus in Non-Saline Potable Water System: A Novel Finding from India.

We present a novel report of abundance of halophilic Vibrio alginolyticus with thermotolerant and enterotoxigenic characteristics from community water system of an inland-focus of India causing diarrheal outbreak as an index pathogen. Though, Vibrio alginolyticus causing diarrhea after exposure to marine water and consumption of seafood was reported globally, its existence in non-saline drinking-water sources with pathogenic viability was unknown. A 'matched-pair-case-control' study identified the primary source of V. alginolyticus infection as 'tap-water' distributed by the municipality, used for drinking (MOR: 8.33; 95% CI 2.51-27.6) and household chores (MOR: 3.75; 95% CI 1.24-11.3). Cardinal toxin gene 'tdh' and other pathogenicity markers viz.tlh, vppC, toxR, VPI, T3SS1 and sxt were detected in V. alginolyticus isolates. Expression potential of the hemolytic genes are demonstrated by hemolysis assay and transcriptome analysis. Altogether 30.55% of isolates exhibited strong hemolytic potential in vitro. RT-PCR revealed uninterrupted virulence gene expression in outbreak strains under heat stress. Surprisingly, ~ 100% of V. alginolyticus from the outbreak focus were sensitive/partially sensitive to all group of antibiotics except β-lactums, carbapenem and quinolones. High drug-sensitivity suggested lack of previous human gut exposure and indicated a fresh dissemination from the environmental niche to the community domain. The maximum likelihood phylogeny depicted multiple clades in V. alginolyticus strains from Pan India sources. Isolated outbreak strains shared common ancestry with the strains from nearby riverine system, a source of 'drinking water' supplied to the affected community, confirming its environmental origin. V. alginolyticus, traditionally a fish-pathogen, is steadily gaining an emerging epidemiological relevance alongside other waterborne diarrheagenic bacteria and its 'thermotolerant' attribute poses additional threat under the canvas of climate change.

One Health Farming: Noninvasive monitoring reveals links between farm vertebrate richness and pathogen markers in outdoor hoofstock

Outdoor farming contributes to biodiversity conservation and enhances animal welfare, but also raises biosafety concerns due to livestock contact with potentially infected wildlife. Thus, there is a need to assess the balance between vertebrate species richness on farms, visits by wildlife species posing a biosafety risk, and pathogen circulation in open-air farming systems. We explored these links in a pilot study involving 15 open-air hoofstock farms (6 cattle, 5 small ruminant, and 4 pig farms), where we conducted interviews and risk point inspections and used two noninvasive tools: short-term camera trap (CT) deployment and environmental nucleic acid detection (ENAD). CTs were deployed to assess the richness of birds and mammals, as well as to determine the percentage of CTs detecting defined risk species. We also collected livestock feces and used sponges to sample surfaces for environmental DNA (eDNA), testing for nine pathogen markers. Total vertebrate richness ranged from 18 to 42 species, with waterholes significantly contributing to farm vertebrate richness, since 48.2 % of all wild vertebrates were detected at waterbodies, and 28.6 % were exclusively detected at waterholes. Pathogen markers detected at risk points correlated with those detected in livestock samples. Notably, the frequency of uidA marker detection correlated with the total number of pathogen markers detected per farm. Overall marker richness, an indicator of pathogen diversity, varied between farms, being higher in small ruminant farms compared to cattle or pig farms. At the farm level, wild vertebrate richness was negatively correlated with the richness of pathogen markers detected at risk points. Additionally, risk points with a higher probability of detecting more pathogen markers had lower vertebrate richness. Although CT-based assessments of vertebrate richness and ENAD-based pathogen marker detection are only indicators of actual biodiversity and farm health, respectively, our findings suggest that farmland vertebrate communities provide important ecosystem services and may help limit the circulation of multi-host pathogens.

Epidemiological and clinical characteristics of cases and detection of pathogens’ markers of tick-borne infections in the Tomsk region

BACKGROUND: Among the spectrum of the tick-borne infections, Lyme borreliosis and tick-borne encephalitis are most often recorded in the Tomsk region, however it is obviously that possible range of the pathogens is not limited to these ones. AIM: To make epidemiological and clinical characteristics of cases and detect frequencies of pathogen’s markers of tick-borne infections among adult patients applying serological and molecular genetic techniques in the Tomsk region during 2020–2023. METHODS: A single-center prospective sample study was conducted applying a comprehensive approach in peripheral blood specimens obtained from 197 patients with suspected tick-borne infections hospitalized during 2020–2023. To verify the diagnoses of tick-borne encephalitis or Lyme borreliosis, specific immunoglobulins of classes M and G to the antigens of virus of tick-borne encephalitis and Borrelia burgdorferi s.l. were determined in paired blood serum samples applying enzyme-linked immunosorbent assay. Additionally, the polymerase chain reaction was used to detect genetic markers of virus of tick-borne encephalitis, B. burgdorferi s.l., B. miyamotoi, Rickettsia spp., Coxiella burnetii, and Francisella tularensis. Epidemiological surveillance data of incidence rates of tick-borne infections obtained from official reports were analyzed as well. MS Office Excel 2010 software was used to calculate frequency morbidity measures. RESULTS: We verified the final diagnosis of tick-borne encephalitis in 52 (26.4%) patients, Lyme borreliosis in 116 (58.9%) patients, co-infection of tick-borne encephalitis and Lyme borreliosis 15 (7.6%) patients, hard tick relapsing fever caused by B. miyamotoi in 8 (4.1%) patients, coinfection of Q fever caused by C. burnetii with tick-borne encephalitis or Lyme borreliosis in 4 (2.0%) patients, and coinfection of tularemia with tick-borne encephalitis or Lyme borreliosis in 2 (1.0%) cases. We identified markers of tick-borne encephalitis and Lyme borreliosis pathogens by polymerase chain reaction in 3.6% and 4.1% of the patients. Most number of individuals with tick bites, and the highest incidence rates of Lyme borreliosis and tick-borne encephalitis were recorded in the suburbs of Tomsk. The age groups of 40–59 years, and elderly people 60 years and older were predominated among the patients. CONCLUSIONS: It was found out that, in addition to Lyme borreliosis and tick-borne encephalitis, there were such tick-borne infections as tick relapsing fever, Q fever, and tularemia in the Tomsk region in the 2020–2023 period.

Influence of Hematocrit and Storage Temperature on the Stability of Dried Blood Samples in Serological Analyses of Tetanus, Diphtheria, and Pertussis.

Dried blood spots (DBSs) enable the study of serological markers of various pathogens without the need to obtain serum/plasma through venipuncture. Sixty-four blood samples were prepared on Whatman™ 903 cards using specimens obtained by venipuncture to study the detection of serological markers of diphtheria, tetanus, and pertussis in DBSs, and their stability 4 months post-collection. An automated chemiluminescent immunoassay was used to investigate diphtheria, tetanus, and pertussis IgG levels from both DBSs and plasma samples. An optimal cut-off value for DBSs was calculated to improve the performance of diphtheria and tetanus serological markers in DBSs, achieving high sensitivity (95% and 98%, respectively) and specificity (91.7% and 92.3%, respectively). No protection against pertussis was found in the population studied. The correlation observed between the plasma and the DBSs processed after sample collection was high (0.967-0.970) for all antibodies studied except pertussis (0.753), both considering hematocrit before sample elution or not. The correlation between DBSs and plasma for diphtheria and tetanus remained strong following a 4-month delay in DBS processing at 4 °C (0.925-0.964) and -20 °C (0.924-0.966), with only a minor decrease observed for diphtheria at room temperature (0.889), while maintaining a strong correlation for tetanus (0.960). For pertussis, the correlation between DBSs and plasma was drastically reduced after delaying its processing for 4 months at any temperature. To summarize, hematocrit levels within the normal range do not affect the processing of DBSs in the study of serological markers of diphtheria, tetanus, and pertussis. The DBS stability for serological diagnosis of diphtheria and tetanus is adequate when samples are stored at -20 °C for a period of 4 months. The pertussis serological marker does not appear to remain stable after 4 months, even when the DBS is stored frozen at -20 °C.

Single-Cell Transcriptome Sequencing Identifies New Molecular Markers for Pathogenicity of IgA Deposition in the Mesangial Region

Single-Cell Transcriptome Sequencing Identifies New Molecular Markers for Pathogenicity of IgA Deposition in the Mesangial Region

CD8+ T cells exacerbate AD-like symptoms in mouse model of amyloidosis

Alzheimer’s disease (AD) is linked to toxic Aβ plaques in the brain and activation of innate responses. Recent findings however suggest that the disease may also depend on the adaptive immunity, as B cells exacerbate and CD8+ T cells limit AD-like pathology in mouse models of amyloidosis. Here, by artificially blocking or augmenting CD8+ T cells in the brain of 5xFAD mice, we provide evidence that AD-like pathology is promoted by pathogenic, proinflammatory cytokines and exhaustion markers expressing CXCR6+ CD39+CD73+/- CD8+ TRM-like cells. The CD8+ T cells appear to act by targeting disease associated microglia (DAM), as we find them in tight complexes with microglia around Aβ plaques in the brain of mice and humans with AD. We also report that these CD8+ T cells are induced by B cells in the periphery, further underscoring the pathogenic importance of the adaptive immunity in AD. We propose that CD8+ T cells and B cells should be considered as therapeutic targets for control of AD, as their ablation at the onset of AD is sufficient to decrease CD8+ T cells in the brain and block the amyloidosis-linked neurodegeneration.

Epidemiological situation of natural focal infections in the Volgograd region in 2022

The article is devoted to the analysis of the epidemiological situation of natural focal infections (NFI) in the Volgograd region in 2022. Materials and methods: The work used the official statistics of the institutions of Rospotrebnadzor on NFI for 2022. The main research method is epidemiological analysis. Results and discussion: It has been established that the numbers of small mammals, potential carriers of NFI, as well as ixodid ticks in the territory of the Volgograd region in the season of 2022 were higher than the same indicators of the previous year (SIPY). The number of mosquitoes, carriers of arbovirus infections, did not exceed the average long-term indicators in any species. When examining environmental objects for the presence of markers of NFI pathogens, a slight increase in the proportion of positive samples compared to SIPY in all nosological forms was noted. Sporadic cases of West Nile fever (WNF), Crimean hemorrhagic fever (CHF), hemorrhagic fever with renal syndrome (HFRS), ixodes ricinus-borne diseases (TBDs), Q fever have been registered; no cases of tularemia, rabies, leptospirosis were detected. The morbidity rates of the population of the Volgograd region for all NFI were at the level or lower than those for the Russian Federation and the Southern Federal District.

Outbreaks of H5N1 High Pathogenicity Avian Influenza in South Africa in 2023 Were Caused by Two Distinct Sub-Genotypes of Clade 2.3.4.4b Viruses.

In 2023, South Africa continued to experience sporadic cases of clade 2.3.4.4b H5N1 high-pathogenicity avian influenza (HPAI) in coastal seabirds and poultry. Active environmental surveillance determined that H5Nx, H7Nx, H9Nx, H11Nx, H6N2, and H12N2, amongst other unidentified subtypes, circulated in wild birds and ostriches in 2023, but that H5Nx was predominant. Genome sequencing and phylogenetic analysis of confirmed H5N1 HPAI cases determined that only two of the fifteen sub-genotypes that circulated in South Africa in 2021-2022 still persisted in 2023. Sub-genotype SA13 remained restricted to coastal seabirds, with accelerated mutations observed in the neuraminidase protein. SA15 caused the chicken outbreaks, but outbreaks in the Paardeberg and George areas, in the Western Cape province, and the Camperdown region of the KwaZulu-Natal province were unrelated to each other, implicating wild birds as the source. All SA15 viruses contained a truncation in the PB1-F2 gene, but in the Western Cape SA15 chicken viruses, PA-X was putatively expressed as a novel isoform with eight additional amino acids. South African clade 2.3.4.4b H5N1 viruses had comparatively fewer markers of virulence and pathogenicity compared to European strains, a possible reason why no spillover to mammals has occurred here yet.

Design and synthesis of hemicyanine-based NIRF probe for detecting Aβ aggregates in Alzheimer’s disease

Alzheimer’s disease (AD), a progressive neurodegenerative disorder, has garnered increased attention due to its substantial economic burden and the escalating global aging phenomenon. Amyloid-β deposition is a key pathogenic marker observed in the brains of Alzheimer’s sufferers. Based on real-time, safe, low-cost, and commonly used, near-infrared fluorescence (NIRF) imaging technology have become an essential technique for the detection of AD in recent years. In this work, NIRF probes with hemicyanine structure were designed, synthesized and evaluated for imaging Aβ aggregates in the brain. We use the hemicyanine structure as the parent nucleus to enhance the probe’s optical properties. The introduction of PEG chain is to improve the probe’s brain dynamice properties, and the alkyl chain on the N atom is to enhance the fluorescence intensity of the probe after binding to the Aβ aggregates as much as possible. Among these probes, Z2, Z3, Z6, X3, X6 and T1 showed excellent optical properties and high affinity to Aβ aggregates (Kd = 24.31 ∼ 59.60 nM). In vitro brain section staining and in vivo NIRF imaging demonstrated that X6 exhibited superior discrimination between Tg mice and WT mice, and X6 has the best brain clearance rate. As a result, X6 was identified as the optimal probe. Furthermore, the docking theory calculation results aided in describing X6′s binding behavior with Aβ aggregates. As a high-affinity, high-selectivity, safe and effective probe of targeting Aβ aggregates, X6 is a promising NIRF probe for in vivo detection of Aβ aggregates in the AD brain.

#2111 Prediction of ESRD in IgA nephropathy using IgA: C3 ratio at diagnosis. 10 year cohort

Abstract Background and Aims IgA Nephropathy is the commonest glomerular pathology with a wide range of outcomes depending on the severity of the circulating serological pathophysiology and on the resulting inflammatory response in the kidney. Prediction of kidney loss, which results in renal failure in 20-40% of cases -versus very benign disease, has largely used traditional clinical parameters such as proteinuria, hypertension and already established renal decline at the point of diagnosis. There has been variable utility from biopsy severity scoring. Pathogenicity of IgA immune complexes may result from activation of complement via the alternative pathway and assessment of serum IgA and complement C3 has been suggested as a marker of pathogenicity. We examined the utility of IgA to C3 ratio in predicting outcomes in a cohort over 10 years Method We examined all the electronic records of biopsy proven IgA cases from 2010 onwards when serum electrophoresis and serum complement became been widely tested at all new renal presentations. These cases were followed to end stage kidney disease or death. A subset of diagnosed patients also had MEST-C score. Results Of 319 biopsy proven IgA cases between 2010 and 2023, 216 had serum IgA and complement studies at diagnosis, with median PCR 309 and median serum creatinine 177. 56 patients had IgA:C3 >4. Outcome of “death or renal replacement therapy” rather than the surrogate “RRT or doubling serum creatinine” was possible due to the length of follow up. Median follow up was 4.5 years, during which 101 (46%) died or reached ESRD. Among those with IgA:C3 > 4, 35 (61%) median survival 5 years, versus 67(42%) of those < 4 (median survival 10 years). Hazard ratio for IgA:C3 >4 was 1.9 (1.2-3.2) Cox Proportional Hazard show this to be independent from presenting creatinine, proteinuria, but not independent of age. A cohort of 107 biopsy proven IgA cases between 2010 and 2015 had data on CMEST score. 62 of this cohort also had IgA and complement measured. There was no correlation with CMEST score, or occurrence of crescents present on biopsy There were no differences in the treatments used following diagnosis of IgA Nephropathy. Similar proportions receiving steroid, calcineurin or mycophenolate. Conclusion IgA to C3 ratio is readily available result which powerfully predicts IgA outcome independently from pathology score and traditional clinical severity estimates. It may well become a useful indicator to target novel immune/complement therapies.

High Diversity of Curtobacterium flaccumfaciens in Poinsettia and Detection of Three Pathogenicity Plasmids for Identification of C. flaccumfaciens pv. poinsettiae

Outbreaks in Europe have raised concerns about poinsettia bacterial canker caused by Curtobacterium flaccumfaciens pv. poinsettiae, described in the United States. Using a semiselective medium containing aztreonam and fosfomycin and selection during isolation based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) spectra, 424 Curtobacterium strains were isolated from Belgian poinsettias of African and European origin. Different populations coexisted: 130 strains were identified as C. flaccumfaciens with scores ≥2.0 and 294 with scores <2.0 or as another Curtobacterium. MALDI-TOF MS libraries constructed using similar medium and extraction procedure and a pathogenicity test on poinsettia were used to screen collections from poinsettia and wheat, a possible alternative host, for C. flaccumfaciens pv. poinsettiae. The concatenated recA- gyrB partial sequences showed that 114 poinsettia or wheat strains belonged to different Curtobacterium species. Eighty-eight nonpathogenic strains and four U.S. strains from litter were intermingled with bean pathogenic C. flaccumfaciens pv. flaccumfaciens strains in the three genetic groups of C. flaccumfaciens but lacked their pathogenicity markers. Four new European recA- gyrB sequatypes of C. flaccumfaciens pv. poinsettiae, obtained from symptomatic and asymptomatic poinsettias, were distinguished from three American sequatypes. Six sequatypes were pathogenic in tests, belonged to genetic groups related to two different genomospecies, and possessed a plasmid. Sequencing of six plasmids revealed three related plasmids containing proteases and a polygalacturonase found only in strains pathogenic in pathogenicity tests and specifically identified by polygalacturonase-based PCR and loop-mediated isothermal amplification assays. Similarities between plant and litter Curtobacterium and the role of plasmids in pathogenicity and punctual transmission of pathogenicity among heterogeneous C. flaccumfaciens are suggested.

The impact of HBx protein on mitochondrial dynamics and associated signaling pathways strongly depends on the hepatitis B virus genotype.

Chronic hepatitis B virus (HBV) infections are strongly associated with liver cirrhosis, inflammation, and hepatocellular carcinoma. In this context, the viral HBx protein is considered as a major factor influencing HBV-associated pathogenesis through deregulation of multiple cellular signaling pathways and is therefore a potential target for prognostic and therapeutic applications. However, HBV-associated pathogenesis differs significantly between genotypes, with the relevant factors and in particular the contribution of the genetic diversity of HBx being largely unknown. To address this question, we studied the specific genotype-dependent impact of HBx on cellular signaling pathways, focusing in particular on morphological and functional parameters of mitochondria. To exclusively investigate the impact of HBx of different genotypes on integrity and function of mitochondria in the absence of additional viral factors, we overexpressed HBx in Huh7 or HepG2 cells. Key signaling pathways were profiled by kinome analysis and correlated with expression levels of mitochondrial and pathogenic markers. Conclusively, HBx of genotypes A and G caused strong disruption of mitochondrial morphology alongside an induction of PTEN-induced putative kinase 1/Parkin-mediated mitophagy. These effects were only moderately dysregulated by genotypes B and E, whereas genotypes C and D exhibit an intermediate effect in this regard. Accordingly, changes in mitochondrial membrane potential and elevated reactive oxygen species production were associated with the HBx-mediated dysfunction among different genotypes. Also, genotype-related differences in mitophagy induction were identified and indicated that HBx-mediated changes in the mitochondria morphology and function strongly depend on the genotype. This indicates a relevant role of HBx in the process of genotype-dependent liver pathogenesis of HBV infections and reveals underlying mechanisms.IMPORTANCEThe hepatitis B virus is the main cause of chronic liver disease worldwide and differs in terms of pathogenesis and clinical outcome among the different genotypes. Furthermore, the viral HBx protein is a known factor in the progression of liver injury by inducing aberrant mitochondrial structures and functions. Consequently, the selective removal of dysfunctional mitochondria is essential to maintain overall cellular homeostasis and cell survival. Consistent with the intergenotypic difference of HBV, our data reveal significant differences regarding the impact of HBx of different genotypes on mitochondrial dynamic and function and thereby on radical oxygen stress levels within the cell. We subsequently observed that the induction of mitophagy differs significantly across the heterogenetic HBx proteins. Therefore, this study provides evidence that HBx-mediated changes in the mitochondria dynamics and functionality strongly depend on the genotype of HBx. This highlights an important contribution of HBx in the process of genotype-dependent liver pathogenesis.

GABAergic interneurons in the hippocampal CA1 mediate contextual fear generalization in PTSD rats.

Fear overgeneralization is widely accepted as a pathogenic marker of post-traumatic stress disorder (PTSD). Recently, GABAergic interneurons have been regarded as key players in the regulation of fear memory. The role of hippocampal GABAergic interneurons in contextual fear generalization of PTSD remains incompletely understood. In the present study, we established a rat model of PTSD with inescapable foot shocks (IFS) and observed the loss of GABAergic interneuron phenotype in the hippocampal cornu ammonis-1 (CA1) subfield. To determine whether the loss of GABAergic interneuron phenotype was associated with fear generalization in PTSD rats, we used adeno-associated virus (AAV) to reduce the expression of GAD67 in CA1 and observed its effect on fear generalization. The results showed that the reduction of GAD67 in CA1 enhanced contextual fear generalization in rats. We investigated whether the PERK pathway was involved in the GABAergic interneuron injury. Increased expression of p-PERK, CHOP, and Caspase12 in GABAergic interneurons of PTSD rats was observed. Then, we used salubrinal, an endoplasmic reticulum stress inhibitor, to modulate the PERK pathway. The salubrinal treatment significantly protected the GABAergic interneurons and relieved fear generalization in PTSD rats. In addition, the results showed that salubrinal down-regulated the expression of CHOP and Caspase12 in GABAergic interneurons of PTSD rats. In conclusion, this study provided evidence that the loss of GABAergic interneuron phenotype in CA1 may contribute to contextual fear generalization in PTSD. The PERK pathway is involved in the GABAergic interneuron injury of PTSD rats and modulating it can protect GABAergic interneurons and constrain contextual fear generalization.

Novel Findings on CCR1 Receptor in CNS Disorders: A Pathogenic Marker Useful in Controlling Neuroimmune and Neuroinflammatory Mechanisms in Parkinson's Disease.

Parkinson's disease (PD) is recognized as the second most common neurodegenerative disease worldwide. Even if PD etiopathogenesis is not yet fully understood, in recent years, it has been advanced that a chronic state of inflammation could play a decisive role in the development of this pathology, establishing the close link between PD and neuroinflammation. In the broad panorama of inflammation and its several signaling pathways, the C-C chemokine receptor type 1 (CCR1) could play a key pathogenic role in PD progression, and could constitute a valuable target for the development of innovative anti-PD therapies. In this study, we probed the neuroprotective properties of the CCR1 antagonist BX471 compound in a mouse model of MPTP-induced nigrostriatal degeneration. BX471 treatments were performed intraperitoneally at a dose of 3 mg/kg, 10 mg/kg, and 30 mg/kg, starting 24 h after the last injection of MPTP and continuing for 7 days. From our data, BX471 treatment strongly blocked CCR1 and, as a result, decreased PD features, also reducing the neuroinflammatory state by regulating glial activation, NF-κB pathway, proinflammatory enzymes, and cytokines overexpression. Moreover, we showed that BX471's antagonistic action on CCR1 reduced the infiltration of immune cells, including mast cells and lymphocyte T activation. In addition, biochemical analyses carried out on serum revealed a considerable increase in circulating levels of CCR1 following MPTP-induced PD. In light of these findings, CCR1 could represent a useful pathological marker of PD, and its targeting could be a worthy candidate for the future development of new immunotherapies against PD.

Functional Analysis of cag Pathogenicity Island Diversity in Helicobacter pylori Isolated from Multi-Ethnic Patients in Malaysia

cagPAI is an important pathogenic marker contributing to disease severity caused by H. pylori infection, encoding proteins for type four secretion systems (T4SS) and the pro-inflammatory cytokines (IL-8) induction in infected host. The heterogeneity of cagPAI genes results in different clinical outcomes in H. pylori infection. The aim of this study was to investigate the functional diversity of cagPAI genes in clinical H. pylori strains isolated from patients with gastroduodenal diseases. Among the isolates investigated, 6 out of 27 (22.2%) harbor intact cagPAI, 3 strains (11.1%) have complete absence of cagPAI, and 18 strains (66.7%) contain a partially deleted island. IL-8 concentration is observed significantly higher among cagPAI-positive isolates (P = 0.045). Of note, IL-8 secretion in the gastric cells infected with H. pylori strains isolated from Malay patients was significantly higher than the gastric cells infected with the strains from non-Malay patients (P = 0.046). The secretion of IL-8 appears in similarly high among intact and partially deleted cagPAI isolates. East Asian cagA isolates induce a notably higher level of IL-8 than that of Western cagA. Inflammatory activities are observed and IL-8 concentration in the milder stage was found in significantly higher quantities than in the severe stage (P = 0.027). The ‘hummingbird’ phenotype was observed to cause severe effects in infected cells with cagPAI-intact H. pylori. The genetic variability of cagPAI isolates from multi-ethnicity patients may have unique implications for disease outcomes with respect to proinflammatory secretion.

Abstract 6825: Epigenetic dysregulation in well-differentiated pancreatic neuroendocrine tumor (PNET): An immunohistochemistry (IHC) analysis

Abstract Epigenetic dysregulation through DNA Methyltransferase 1 (DNMT1) is integral to pancreatic neuroendocrine tumorigenesis. We aim to investigate the interplay between epigenetic modifiers and pathogenetic markers in the PNET microenvironment. 12 paraffin-embedded PNET specimens and 12 benign adjacent tissue specimens, used as controls, were obtained from Cooper University Hospital Pathology Department (IRB 23-130). IHC was performed using epigenetic markers, including DNMT1, DNMT3A, DNMT3B, and 5-Hydroxymethylcytosine (5-HMC), and pathogenic markers, including Multiple Endocrine Neoplasia 1 (MEN1), Protein-Tyrosine Phosphatase (PTEN), and Ras Association Domain Family 1 (RASSF1). Image J software was utilized for IHC scoring under the guidance of a board-certified pathologist. DNMT1 was upregulated in PNET tumor specimens with a mean density score of 72.02 +/- 31 versus 16.02 +/- 17 in adjacent benign tissue (p<.001). Grade 2 tumors displayed increased DNMT1 expression compared to Grade 1 tumors with a mean score of 37 +/-21 versus 71 +/- 23 (p<.05). Decreased expression of 5-HMC was noted in tumor specimens in comparison to benign tissue with a mean score of 3.92 +/- 5.05 versus 19.37 +/- 5.05 (p<.05). Loss of 5-HMC was observed in higher tumor grades with a mean score of .79 +/- 2 versus 8.2+/-2 (p<.01). DNMT3B was upregulated in PNET tumor specimens with a mean score of 13.16 +/- 3.94 versus 7.02+/- 2.553 (p<.01) in adjacent benign tissue and was positively correlated to DNMT1 expression. DNMT3A was not found to have a difference in expression between tumor and benign tissue. MEN1 was downregulated in PNET, with 45% of tumor specimens displaying loss of MEN1 expression. Mean MEN1 density in tumor tissue was 4.47 +/- 5.95 versus 11.41 +/- 3.53 in adjacent benign (p<.01). Increased expression of DNMT1 found to negatively correlate with MEN1 expression (R2=.23). Loss of RASSF1 expression was present in 70% of tumor specimens with a mean score of 1.32 +/- 1.99 versus 9.50+/- 6.89 (p<.001) in adjacent benign tissue. RASSF1 expression correlated negatively with DNMT1 (R2=.21). PTEN expression showed intratumoral homogeneity to benign tissue, and its expression was not correlated with any epigenetic markers. Our work provides a framework of epigenetic regulators involved in the pathobiology of PNET, and can refine prognostic evaluation while providing therapeutic benefits for patients with PNET. Citation Format: Zena Saleh, Yazid Ghanem, Hansa Joshi, Mathew C. Moccia, Helen Toma, Yahui Li, Upsana Joneja, Tao Gao, Young Hong. Epigenetic dysregulation in well-differentiated pancreatic neuroendocrine tumor (PNET): An immunohistochemistry (IHC) analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6825.

Role of ixodid ticks in tick-borne pathogen spread and circulation in the Belarusian Lakeland

Results of ixodid tick analysis for their ecological, epizootological and epidemiological significance for tick-borne pathogen spread across the Belarusian Lakeland are presented. The ticks were collected in publicly accessible areas of the Vitebsk Raion in April – November 2022: 8 routes were tracked, 18 flag-km were passed, 529 tick specimens were collected, including 350 imago ticks and 179 nymph ticks. The ixodid tick genus and species were determined using N. A. Filippova’s ixodid tick determinator. All caught ticks were tested for Borrelia spp., Anaplasma spp. (Ehrlichia spp.), Babesia spp. and Tick-borne encephalitis virus genetic materials with real-time polymerase chain reaction using the reagent kit for nucleic acid extraction from environmental samples in accordance with the manufacturer’s instruc­tions. The specimens were grouped in accordance with the MG 3.1.1027-01 “Collection, recording and preparation for laboratory tests of blood-sucking arthropods being vectors of natural focal infections”; therewith, one specimen includes only one tick. Differences in the numbers of ixodid ticks and the occurrence of genetic markers of tick-borne pathogens in them were found to be associated with ecological characteristics of the examined territories. The following epidemically and epizootically significant ticks contributing to transmissible infection and invasion spread were found in the Belarusian Lakeland: ticks of Ixodes and Dermacentor genera; their frequency index was 70.1 and 29.9%, respectively. Tick-transmitted pathogen prevalence rate in the examined territories of the Vitebsk Raion was as follows: 61.7% for Borrelia spp., 25.8% for Anaplasma spp. (Ehrlichia spp.) and 25% for Babesia spp., mixed infections were found in 10.8% of the ticks. No tick-borne encephalitis virus genetic materials were found in the specimens. Total infection rate for ixodid ticks was 22.7%.

Epidemiological situation of natural focal infections in the Volgograd region in 2023

Objective: analysis of epizootiological and epidemiological aspects of natural focal infections, namely plague, tularemia, HFRS, West Nile fever, Crimean hemorrhagic fever, Ixodic tick-borne borreliosis, Cu fever, leptospirosis and rabies, in the Volgograd region over 10 months in 2023.Materials and methods: the article analyzes the epidemiological manifestations of natural focal infections (NFI) in the Volgograd region for 10 months of 2023. The work uses official statistical data from institutions of Rospotrebnadzor of the Volgograd region on NFI for 2023. The main research method is epidemiological analysis.Results: the numbers of small mammals, potential carriers of NFI, on the territory of the Volgograd region in the 2023 season were within the same indicators of the previous year (SIPY), while the number of ticks in the spring-summer period was higher than the SIPY, but in the fall the number of arthropods in stationary points was lower than in previous seasons. The number of mosquitoes carrying arbovirus infections exceeded the long-term average. When studying environmental objects for the presence of markers of NFI pathogens, a slight decrease in the proportion of positive samples was noted compared to SIPY for all nosological forms. Over the 10 months of this year, there has been a deterioration in the epidemiological situation in the Volgograd region for several infections compared to the same period in 2022. There were 18 cases of West Nile fever (WNF), 10 cases of ixodid tick-borne borreliosis (ITB), and 6 cases of hemorrhagic fever with renal syndrome (HFRS). No patients with plague, tularemia, rabies, leptospirosis, Crimean hemorrhagic fever (CHF) and Q fever were identified.Conclusion: the presence of natural foci of dangerous infectious diseases of varying degrees of activity on the territory of the Volgograd region requires constant monitoring and an adequate response to changes in the foci in order to prevent complications of the epidemiological situation due to NFI.

Differential Expression of Noncoding RNAs Revealed Enhancer RNA AC016735.2 as a Potential Pathogenic Marker of Congenital Microtia Patients.

Congenital microtia is a complex maxillofacial malformation with various risk factors. This study aimed to find potential pathogenic noncoding RNAs for congenital microtia patients. We collected 3 pairs of residual ear cartilage samples and corresponding normal ear cartilage samples from nonsyndromic congenital microtia patients for microarray experiments. The differentially expressed RNAs were screened, and enrichment analysis and correlation expression analysis were performed to elucidate the function of the differentially expressed genes (DEGs). We further investigated the most significantly differentially expressed long noncoding RNA (lncRNA), AC016735.2, through follow-up analyses including RT-qPCR and Western blotting, to validate its differential expression in residual ear cartilage compared with normal ear cartilage. SiRNA was designed to study the regulatory role of AC016735.2, and cell proliferation experiments were conducted to explore its impact on residual ear chondrocytes. Analysis of the microarray data revealed a total of 1079 differentially expressed RNAs, including 305 mRNAs and x lncRNAs, using a threshold of FC>1.5 and P<0.05 for mRNA, and FC>1.0 and P<0.05 for lncRNA. Enrichment analysis indicated that these genes are mainly involved in extracellular matrix regulation and embryonic development. AC016735.2 showed the highest differential expression among the eRNAs, being upregulated in residual ear cartilage. It acts in cis to regulate the nearby coding gene ZFP36L2, indirectly affecting downstream genes such as BMP4, TWSG1, COL2A1, and COL9A2. Significant differences were observed in the expression of lncRNAs and mRNAs between residual ear cartilage and normal auricular cartilage tissues in the same genetic background of congenital microtia. These differentially expressed lncRNAs and mRNAs may play crucial roles in the occurrence and development of microtia through pathways associated with extracellular matrix regulation and gastrulation. Particularly, AC016735.2, an eRNA acting in cis, could serve as a potential pathogenic noncoding gene.

Regulation of Onecut2 by miR-9-5p in Japanese encephalitis virus infected neural stem/progenitor cells.

MicroRNAs have emerged as key disease pathogenic markers and potential therapeutic targets. In this study, we solidify this concept by studying a key miRNA, miR-9-5p, in Japanese encephalitis virus infection of neural stem/progenitor cells. miRNA target Onecut2 has a possible role in stem cell pool biology. Here, we show a possible mechanistic axis worth investing in neurotropic viral biology.