Viral Markers Research Articles

HBV shows different levels of adaptation to HLA class I-associated selection pressure correlating with markers of replication

Background & AimsImmune responses by CD8 T cells are essential for control of HBV replication. Although selection of escape mutations in CD8 T cell epitopes has been previously described in HBV infection, its overall influence on HBV sequence diversity and correlation with markers of HBV replication remain unclear. MethodsWhole-genome sequencing was applied to HBV isolates from 532 patients with chronic HBV infection and high-resolution HLA class I genotyping. Using a Bayesian model (HAMdetector) for Identification of HLA-associated mutational states (HAMs) the frequency and location of residues under CD8 T cell selection pressure were determined and the levels of adaptation of individual isolates were quantified. ResultsUsing previously published thresholds for the identification of HAMs, a total of 295 residues showed evidence of CD8 T cell escape, the majority of which were located in previously unidentified epitopes. Interestingly, HAMs were highly enriched in the HBV core protein compared to all other proteins. When individual HBV isolates were compared, different levels of adaptation to HLA class I immune pressure were noted. The level of adaptation increased with patient age and correlated with markers of replication, with low levels of adaptation in HBeAg-positive infection. Furthermore, the levels of adaptation negatively correlated with HBV viral load and HBsAg levels, consistent with high levels of HLA class I-associated selection pressure in patients with low replication level. ConclusionsHBV sequence diversity is shaped by HLA class I-associated selection pressure with the HBV core protein being a predominant target of selection. Importantly, different levels of adaptation to immune pressure were observed between HBV infection stages, which need to be considered in the context of T cell-based therapies. Impact and implicationsThe immune response mediated by CD8 T cells plays a critical role in controlling HBV infection and shows promise for therapeutic strategies aimed at achieving a functional cure. This study demonstrates that mutational escape within CD8 T cell epitopes is common in HBV and represents a key factor in the failure of immune control. Notably, the HBV core protein emerges as the primary target of CD8 T cell selection pressure. Additionally, the observed correlation between HBV adaptation levels and viral replication markers indicates that CD8 T cell immunity may influence transitions between phases of chronic HBV infection.

Afferent and Efferent Connections of the Postinspiratory Complex (PiCo) Revealed by AAV and Monosynaptic Rabies Viral Tracing.

The control of the respiratory rhythm and airway motor activity is essential for life. Accumulating evidence indicates that the postinspiratory complex (PiCo) is crucial for generating behaviors that occur during the postinspiratory phase, including expiratory laryngeal activity and swallowing. Located in the ventromedial medulla, PiCo is defined by neurons co-expressing two neurotransmitter markers (ChAT and Vglut2/Slc17a6). Here, we mapped the input-output connections of these neurons using viral tracers and intersectional viral-genetic tools. PiCo neurons were specifically targeted by focal injection of a doubly conditional Cre- and FlpO-dependent AAV8 viral marker (AAV8-Con/Fon-TVA-mCherry) into the left PiCo of adult ChatCre/wt: Vglut2FlpO/wt mice, for anterograde axonal tracing. These experiments revealed projections to various brain regions, including the Cu, nucleus of the solitary tract (NTS), Amb, X, XII, Sp5, RMg, intermediate reticular nucleus (IRt), lateral reticular nucleus (LRt), pre-Bötzinger complex (preBötC), contralateral PiCo, laterodorsal tegmental nucleus (LDTg), pedunculopontine tegmental nucleus (PPTg), periaqueductal gray matter (PAG), Kölliker-Fuse (KF), PB, and external cortex of the inferior colliculus (ECIC). A rabies virus (RV) retrograde transsynaptic approach was taken with EnvA-pseudotyped G-deleted (RV-SAD-G-GFP) to similarly target PiCo neurons in ChatCre/wt: Vglut2FlpO/wt mice, following prior injections of helper AAVs (a mixture of AAV-Ef1a-Con/Fon oG and viral vector AAV8-Con/Fon-TVA-mCherry). This combined approach revealed prominent synaptic inputs to PiCo neurons from NTS, IRt, and A1/C1. Although PiCo neurons project axons to the contralateral PiCo area, this approach did not detect direct contralateral connections. We suggest that PiCo serves as a critical integration site, projecting and receiving neuronal connections implicated in breathing, arousal, swallowing, and autonomic regulation.

Hepatocyte nuclear factor 1 alpha variants as risk factor for hepatocellular carcinoma development with and without diabetes mellitus

BackgroundHNF1A gene variants have been reported to be involved in developing mature onset diabetes mellitus (DM). Many studies reported the role of DM as a risk factor for hepatocellular carcinoma (HCC) development. To date, it has not been reported whether HNF1A gene variants are associated with the risk of DM in cirrhotic patients and their subsequent HCC. ObjectiveTo evaluate the HNF1A (the hepatocyte nuclear factor 1 homeobox A) genetic variants as a cofactor with DM for HCC development in hepatitis C virus (HCV)-infected patients. Subjects and methodsThis study was conducted on 140 subjects; 30 had HCC without DM, 30 HCC with DM, and 40 patients had DM with no HCV infection or had HCC; in addition, 80 healthy volunteers with matched ages and genders were enrolled in the study as a control group. Liver function tests, hepatitis viral markers, alpha-fetoprotein (AFP), fasting sugar and HBA1c and HNF1A (rs2464196 and rs1169310) using real-time polymerase chain reaction (PCR) were done for all participants. ResultsThe frequency of HNF1A rs2464196 (AA) genotype in patient groups (DM, HCC, HCC + DM) was significantly higher compared to the control group (P = 0.006, P = 0.018, P < 0.001 respectively). The combined dominant model (AA + GA) of rs 2,464,196 was significantly higher than the (GG) genotype in patient groups (DM, HCC, HCC + DM) than the control group. In addition, the frequency of the AA genotype is more prevalent in HCC + DM (73 %) compared to the group of DM or HCC patients. In contrast, the HNF1A rs1169310 (TT, TC or CC genotypes) showed no significant difference among the four studied groups and their T or C allele distributions. ConclusionThis finding suggested that the HNF1A rs2464196 (AA) genotype could be associated with DM and may raise the possibility of HCC development among HCV-infected patients who harbour this genotype more than (GG). On the contrary, the HNF1A rs1169310 polymorphism was of no significance as a risk factor in the current study. However, as we limited our study to Egyptian participants, more research on other ethnic groups would be required. Also, large scale studies are recommended on other variants of HNF1A to clarify the role of this gene in HCC development.

Prevalence and risk factors for blood-borne viruses among multi-transfused patients in Sana'a City, Yemen

Background: Blood-borne viruses (BBVs), like hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV) among multi-transfused patients (MTPs), remain a major public health problem in Yemen. This study aimed to determine the seroprevalence of BBVs and associated risk factors among MTPs. Methods: A cross-sectional study was conducted on MTPs at the Yemeni Society for Thalassemia, National Oncology Center, and Al Kuwait University Hospital in Sana’a City. Data were collected through face-to-face interviews using a predesigned questionnaire. Blood samples were drawn and tested for HBsAg, anti-HCV, and HIV-1,2 by using an electrochemiluminescence immunoassay technique. Results: Among 361 MTPs, the overall seroprevalence of BBVs was 6.8%; seropositivity of HBsAg, anti-HCV, and anti-HIV-1,2 was 3.9%, 2.9%, and 0.0%, respectively. An increase in family size and low income were significantly associated factors with HBV infection. In addition, increases in age and blood transfusion units were found to be significant predictors for HCV (p&lt;0.05). Conclusion: Although the seroprevalence of BBVs is low, it remains a major problem among MTPs. The most frequent infection was HBV, followed by HCV, with a significantly higher rate among leukemia patients. Regular mentoring for viral markers and mandatory implementation of HCV antigen-antibodies testing as well as advanced technology in blood screening are highly recommended.

Effect of interleukin 1b inhibition with canakinumab on inflammation and viral persistence in people with human immunodeficiency virus

Abstract Introduction Effectively treated people with HIV (PWH) have elevated risk of cardiovascular disease (CVD). Inflammatory markers are predictive of CVD and mortality among PWH. The role of the myeloid system in driving inflammation and atherosclerosis has been recently recognized. Canakinumab (a monoclonal antibody to IL-1β) reduces inflammation and CVD events among people with elevated hsCRP after myocardial infarction without HIV. We evaluated the impact of IL-1β inhibition using canakinumab on HIV parameters, inflammation, HIV persistence markers, arterial inflammation, and hematopoietic activity in PWH. Methods PWH on effective ART who had CVD or were at risk for CVD were randomized 2:1 to receive 150 mg of canakinumab subcutaneously or matched placebo at weeks 0 and 12. Arterial inflammation and bone marrow metabolic activity were assessed using F18 FDG PET/CT at baseline and week 18. T cell and monocyte activation were evaluated using multiparameter spectral cytometry. The viral reservoir was quantified using Tat/rev Induced Limiting Dilution Assay (TILDA), HIV DNA and cell-associated RNA. The primary endpoints were change in CD4 and CD8 count. We assessed group effects over time using linear mixed effects models. Results 33 individuals were randomized (median age of 60 years old (IQR 57.5, 64.5) and 94% male); 25 received canakinumab and 8 received placebo. There were no significant differences at baseline. As expected, IL-1β increased among those treated with canakinumab at weeks 4- 24 (p&amp;lt;0.01 for each) and returned to baseline at week 36. There was one death from sepsis in an individual aged 84 with CVD and poorly controlled diabetes who received canakinumab. There were no significant changes in CD4 count, CD8 count, platelet count, creatinine, AST, or ALT by group. Treatment was not associated with a greater reduction in arterial inflammation in the most diseased segment compared to placebo (-6.9%; 95% CI -30.4 to 24.5%; p=0.62). However, bone marrow metabolic activity decreased in the canakinumab group versus placebo (-14.4%, 95% CI -26.5% to -0.2%; p=0.047). Treatment was associated with increased CD163 expression on monocytes at weeks 24 (p=0.03) and 36 (p&amp;lt;0.001), and lower caspase activity at week 36 (p=0.02). There was expansion of anti-inflammatory CD16+ patrolling monocytes that co-express CD163+CX3CR1+ and a decrease in pro-inflammatory CD16+ patrolling monocytes that are Caspase1+CCR2+. Treatment was not associated with differences in hsCRP, IL-6, IL-16, IL-18, MCP-1, sCD14, sCD163, T cell subsets, NK cell subsets, or viral persistence markers. Conclusion Among treated PWH, targeted inhibition of IL-1β using canakinumab results in decreased bone marrow metabolic activity and a shift toward anti-inflammatory monocyte populations. These findings shed light on mechanisms underlying how targeted IL-1β inhibition using canakinumab prevents CVD events by altering monocyte populations and reducing systemic inflammation.

A case of complete response to radiotherapy combined with durvalumab and tremelimumab in a patient with unknown primary hepatocellular carcinoma arising in the lumbar spine.

A 58-year-old man visited an orthopedic clinic complaining of pain in his right lower back and numbness in his lower limbs for one month. Imaging tests revealed a tumorous lesion from the left side of the second lumbar vertebra to the paraspinal muscles. CT-guided biopsy of the tumor was performed, and immunostaining results diagnosed hepatocellular carcinoma (HCC). Although the liver showed signs of chronic liver damage, no primary tumor was found within the liver or in other organs. Blood tests showed negative hepatitis virus markers for both HBV and HCV. The tumor markers AFP, AFP-L3, and DCP were high. Because he developed spinal cord compression syndrome due to a lumbar tumor, radiation therapy and denosumab administration were performed. Subsequently, systemic therapy with durvalumab plus tremelimumab was started. In the year following the start of treatment, the tumor has shrunk, and no new lesions have been observed. Tumor markers have also decreased. We have experienced a case of HCC in the lumbar spine without a primary tumor in the liver. This is a very rare case, and the combination therapy with durvalumab and tremelimumab resulted in a complete response, which we consider to be a valuable case.

Prevalence & clinical outcome of autoimmune encephalitis versus viral encephalitis in children with acute encephalitis syndrome: A prospective observational study.

Background & objectives Acute encephalitic syndrome (AES), encompasses a wide spectrum of potential causes, clinical presentations, and outcomes. While infectious encephalitis is generally considered more prevalent, autoimmune encephalitis is emerging as a significant aetiology. Neuronal autoantibodies have been identified independently or in association with acute viral encephalitis. The primary objective of this study was to ascertain the prevalence and clinical manifestation of autoimmune encephalitis as well as of coexisting viral markers in children with AES. Methods This study was a prospective observational investigation conducted in a hospital setting. It involved enrolling children with AES who were admitted to specific tertiary hospitals. Children were subjected to examinations to detect the presence of viral markers and neuronal autoantibodies in both their blood and cerebrospinal fluid (CSF). All the participants received treatment based on established guidelines and was followed for six months for outcome assessment. Results During the study period, 867 children with AES were examined. Among these cases, 37 children (4.2%) were diagnosed with autoimmune encephalitis, and all of them tested positive for anti-NMDAR (N-methyl-D-aspartate receptor) antibodies. Evidence of viral infection was seen in 409 (47.1%) of cases, out of which nearly 254 (29.2%) children had detectable HSV IgM antibodies. Among the 37 children with autoimmune encephalitis, 25 (67.5%) had evidence of a viral trigger, with eight of them tested positive for HSV IgM antibodies. The clinical presentation of autoimmune-associated AES was similar to those with viral aetiology. Interpretation & conclusions Autoimmune encephalitis triggered by neurotropic (HSV) viral infection was more prevalent in this study than in the earlier reports. Typically, these children show positive responses to immunosuppressive treatments if administered promptly. It is hence advisable to assess children who exhibit behavioural issues and movement disorders for possible autoimmune encephalitis.

Persistence of respiratory, enteric, and fecal indicator viruses in fecal sludge from on-site sanitation in Dakar, Senegal

ABSTRACT As wastewater-based epidemiology (WBE) broadens to include prevalent diseases with significant global health impact, existing surveillance systems concentrate on sewer-based infrastructure, which excludes the 2.7 billion people using non-sewered systems. To address this gap, our study explores the potential of fecal sludge treatment plants for WBE, emphasizing the stability of virus RNA targets within pooled fecal sludge. We screened fecal sludge from a centralized treatment facility in Dakar, Senegal for SARS-CoV-2, human norovirus, and microbial source trackers pepper mild mottle virus and tomato brown rugose fruit virus (ToBRFV). Decay kinetics of viral genomic RNA markers were examined at 4, 15, and 30 °C over 70 days. Results indicate the high persistence of viral targets in fecal sludge (T90 value of 3.3 months for exogenous SARS-CoV-2 N1 and N2 and 6.2 months for ToBRFV), with all targets detected throughout the 70-day experiment under various temperatures with limited decay (&amp;lt;90% reduction). This study addresses a crucial gap in understanding virus persistence in on-site sanitation systems by providing essential decay rate constants for effective target detection. Our results indicate that sampling at centralized facilities treating fecal sludge from on-site sanitation could facilitate localized pathogen surveillance in low-income settings.

Trends of coinfections among healthy blood donors: COVID-19 pandemic repercussion

ABSTRACT Background: Human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV) coinfection has emerged as a leading cause of morbidity throughout the world in the last two decades. The coronavirus disease 2019 (COVID-19) pandemic has escalated the disease burden further by increasing the number of intravenous (IV) drug abusers and unemployment. Aim: The present study was done to analyse the impact of COVID-19 on seroprevalence as well as trends during pre, post and pandemic years of coinfection and mono-infections in the Malwa region of Punjab. Setting and Design: This descriptive cross-sectional study was done in the department of immunohematology and blood transfusion at a tertiary care hospital. Materials and Methods: The data on transfusion-transmitted infections (TTIs) was collected for a period of four years from 2019 to 2022, that is, pre, post and during the pandemic period. All the blood samples were screened for viral markers, HIV I and II, HCV, and hepatitis B surface antigen (HBsAg) using the enzyme-linked immunosorbent assay (ELISA) technique. Malarial antigen and syphilis infection testing was done using a rapid diagnostic card test. The total number of sero-reactive cases and their distribution were noted. Results: A total of 58,953 donors were screened and included during the study period. Each blood donor was identified by a donor registration number. The overall TTI seroprevalence in blood donors was 2.83% (n = 1670). The seroprevalence of TTIs in blood donors showed an increasing trend for HIV, HCV, and HBsAg in 2019–2021, whereas there was a decrease in reactivity status in the 2022 (back to pre-pandemic year). There was a significant increase in the coinfection rate from 0.1% to 0.25%. Conclusion: The COVID-19 pandemic impact on the coinfection rate of TTI was significant. To curb these TTIs and coinfections, education and public awareness are the key factors. India is a developing country, so transfusion medicine specialists need to work day and night to practice safe blood services.

Impact of membrane surface and module damage on virus removal and integrity in RO membranes

RO membrane modules are very effective in removing viruses and salts, but concerns remain regarding their long-term integrity in full-scale RO spiral wound systems. Membrane defects can arise from delamination, chlorine, or abrasive particle exposure, and module defects can arise from permeate tube, O-ring, and glue line damage. These issues compromise the membrane and module's performance, allowing viruses to pass that are not detected by conductivity monitoring. This paper assesses the impact of damage on virus removal using biological indicators (natural virus markers (NV)) and non-biological surrogates (salt, sulfate, rhodamine WT, pyranine). Three categories of module damage were studied and characterized using different autopsy techniques: membrane module component damage (permeate tube damage, O-ring damage, and membrane delamination), oxidative membrane damage (by hypochlorite exposure dose), and membrane surface damage (by abrasive particle exposure). Module component damage resulted in increased water permeability and decreased natural virus rejection. The conductivity remained similar to that of an intact module except in the case of permeate tube damage. Chlorine exposure (9000 ppm.h) didn't result in detectable NV markers in the permeate, indicating an intact support layer despite active layer damage. Abrasive particle exposure by suspended silicon carbide in the feed resulted in scratches, observed by SEM images, and a decline in the rejection of fluorescent marker, which indicates membrane surface damage. Results demonstrate that NV markers most consistently determine virus removal capacity in modules compromised by component damage. However, for assessing active layer damage, solutes like rhodamine-WT and pyranine prove more effective. This underscores the necessity of employing multiple indicators for a comprehensive evaluation of membrane integrity.

Mass screening of hepatitis B and C in Burkina Faso: seroprevalence update and impact of hepatitis B vaccine.

Updated data on the seroprevalences of hepatitis B and C viruses (HBV, HCV) are required to enable the adaptation of control strategies. In this study, we aimed to: (i) estimate the seroprevalences of HBsAg carriers and HCV exposure in the general population, and (ii) determine the impact of vaccination on HBV circulation since its introduction in 2006 in the Expanded Program on Immunization (EPI). From October 2020 to October 2022, a mass screening campaign was conducted in 10 cities across Burkina Faso. Individuals of all ages and genders who consented to participate were screened for viral markers (HBsAg, anti-HCV) using rapid diagnostic tests. The proportions of HBsAg carriers and HCV exposure were calculated using Stata, and logistic regression was used to assess the impact of HBV vaccination on HBsAg carriage. A total of 15,650 participants were enrolled in the study. Of these, 51.4% were women and the age range was from 1 to 97 years. All participants were screened for HBsAg and 7,507 were also screened for anti-HCV. Overall, the seroprevalence of HBsAg was 8.8% and 2.6% for anti-HCV. The results indicated that age, gender, and place of residence were associated with HBV infection. The prevalence of HBV and HCV infections remains high in Burkina Faso. Prevention strategies, including initial mass screening with rapid diagnostic tests and vaccination, need to be intensified.

A Study of Cadaveric Skin Graft Harvest and Usage: An Observational Prospective Pilot Study.

Background Burn injuries pose a significant global public health challenge, causing substantial morbidity and mortality, particularly in low- and middle-income countries. Timely and effective wound coverage is critical in treating severe burns to prevent infection, reduce pain, and promote healing. Cadaveric skin grafts (allografts) have become essential in treating extensive burn wounds, serving as temporary biological dressings to prepare the wound bed for autografting. This study aims to comprehensively analyze the process of cadaveric skin graft harvest and usage in a tertiary care setting. It seeks to evaluate the procedures, challenges, and outcomes associated with cadaveric skin grafts, contributing to optimizing burn care practices and improving patient outcomes. Methods This observational study was conducted at a tertiary care hospital and burns center with a skin bank, involving 44 cadaveric skin graft harvests and 27 applications between July 1, 2011, and June 30, 2013. The study focused on prospective donors and recipients needing cadaveric skin grafts. Inclusion criteria for donors included consent from the next of kin and the absence of infections or septicemia, while exclusion criteria included prolonged post-mortem intervals and medico-legal cases. The procedures adhered to the Euro Skin Bank protocols, encompassing retrieval, processing, storage, and usage. Data were analyzed using Epi-Info version 7.2.1 software, employing descriptive statistics for categorical variables. Ethical clearance was obtained from the university ethical committee, with mandatory written informed consent for skin donation. Results Out of 519 deaths in the tertiary care hospital, significant barriers to skin donation included septicemia, skin changes, late reporting, young age, medico-legal issues, and positive viral markers. Notably, 114 (21.97%) of next of kin refused consent. Cadaveric skin was harvested in five (11.36%) cases, with potential donors identified in 78 (15.02%) of deaths. Donors were predominantly older males, with efficient procurement processes ensuring timely harvests. The tertiary burns center facilitated 39 (88.63%) cases of cadaveric skin harvests with a skin bank, either at the donor's home or other hospitals notified to the burns center. Cadaveric skin grafts were applied in 27 cases, primarily for burns, with high graft uptake observed over 10 days. Non-healing ulcers showed 100% graft uptake. The survival rate among burn patients was 20 (74%), with deaths mainly due to sepsis and multi-organ failure. Significant barriers to obtaining consent included a lack of awareness, superstitions, social stigmas, and religious objections. Conclusion The study highlights the critical role of cadaveric skin in managing extensive wounds, particularly burns. Despite challenges in obtaining consent and limited donor availability, cadaveric skin grafts effectively prepared wound beds for autografting, contributing to improved patient outcomes. Increasing community awareness and addressing superstitions and social stigmas are essential for improving donation rates.

Abstract B049: Oncolytic virotherapy as an effective tool to synergistically enhance therapeutic efficacy of standard systemic treatments for pancreatic cancer

Abstract Background: Standard therapies provide marginal benefits in overall survival and leading immunologic agents have proven ineffective in the immunologically cold environment of pancreatic ductal adenocarcinoma (PDAC). Current standard of care chemotherapies experience demonstrable resistance in the clinical setting with primary and secondary resistance frequently leading to poor outcomes. Oncolytic virus (OV) therapies are emerging as an avenue of therapeutic potential with vesicular stomatitis virus (VSV) providing an onco- selective platform with potential for rapid replication and minimal outside virulence. The aim of this project is to determine if the addition of OV to the standard systemic treatments would result in a synergistic improvement of the therapeutic efficacy, and investigate the potential mechanisms involved. Methods: A VSV variant containing the Morreton G protein inserted into a wild-type VSV backbone (VMG)was propagated and purified for in vitro and in vivo studies. In vitro studies investigating oncolytic potential and viral kinetics of VSV in combination with FOLFIRINOX (FFX) and Gemcitabine Paclitaxel (GP) therapies were compared to monotherapy in multiple cell culture formats including 2D Monolayer, 3D Spheroid, and ongoing investigation into patient derived organoids. Proteomics analysis of monotherapies along with combination therapy was performed on in vitro cultures. Ongoing in vivo studies are investigating the therapeutic potential of concurrent treatments. Results: Cytotoxicity assay studies of equivalent multiplicities of infection (MOI) demonstrated increased cytotoxicity of combination therapy (Percent viable 10.2% ± 1.3%) compared to virus alone (30.7% ± 1.9%) or FFX alone (84.4% ± 6.6%) in Hs766T PDAC cell lineAsPC-1 cell line demonstrated similar effects with minimal cytotoxicity from FFX alone (100% ± 5%) with comparable resistance to virus alone (69.9% ± 1.8%) in comparison to combination therapy (40.3% ± 1.0%). Additional cell lines including Panc-01 and MiaPaCa2 provided similar results. Treatment of formed spheroids with combination therapy resulted in reduction in overall size of spheroid along with increased Viral GFP marker fluorescence, in combination group, compared to monotherapy with each treatment. Proteomic analysis investigating concomitant treatment with both standards of care produced unique expression profiles. Multiple pathways including cell cycle regulation, apoptosis, MAPK signaling, PI3K-Akt signaling, ErbB signaling showed significant regulatory changes in protein expression across all treatment groups. Conclusion: VSV oncolytic virus in conjunction with standard of care therapy demonstrates a potentially effective approach for synergistic enhancement of the therapeutic efficacy against PDAC. Ongoing research is investigating the immunomodulatory and metabolic effects of both monotherapies in conjunction with combination therapy to further determine the synergistic mechanisms involved and how they can be further utilized to overcome treatment resistance in PDAC. Citation Format: Conner Hartupee, Amirsalar Mansouri, Joycelynn Coleman-Barnett, Dana Adamcova, Dorota Wyczechowska, Bolni Marius Nagalo, Mulu Tesfay, Jovanny Zabaleta, Luis Del Valle, John West, Jiri Adamec, Omeed Moaven. Oncolytic virotherapy as an effective tool to synergistically enhance therapeutic efficacy of standard systemic treatments for pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr B049.

Molecular Epidemiology of Hepatitis D Virus in the North-East Region of Romania.

The hepatitis D virus (HDV) superinfection of individuals with chronic hepatitis B virus (HBV) infection causes severe liver damage and the poorest long-term prognosis among viral hepatitis. This is attributed to the unique pathogenic mechanisms of HDV characterized by a direct cytopathic effect on hepatocytes and a significant impairment of the host immune response. The HDV genotype largely influences the extent of the pathogenic mechanisms with consequences on disease progression towards cirrhosis, liver decompensation, or hepatocellular carcinoma. In this context, identifying the circulating HDV genotypes in European regions with high prevalence, such as Romania, is crucial for effectively managing the long-term liver health. Here, we report the first comprehensive HDV study in Romania that clinically characterizes 82 patients and performs HDV genotyping by combining the nested-PCR reaction with sequencing analysis in 49 samples with an HDV-RNA load higher than 5000 IU/mL. While all isolates in our study belong to the HDV-1 genotype, the phylogenetic analysis based on sequence data from GenBank reveals the presence of the following potential three groups: (i) Italy and France; (ii) Spain; and (iii) Turkey, Iran, Pakistan, and Germany. This broad clustering highlights the recent surge in migration to and from Western Europe and the Middle East. Equally important, no differences in viral markers, clinical and paraclinical parameters, or treatment options were observed between these identified clusters. Nevertheless, this study considerably advances the understanding of hepatitis D epidemiology and clinical aspects in Romania.

An unusual cause of liver neoplasm in an older female.

We presented a 66-year-old woman with T2DM who had a liver mass discovered incidentally during hospitalization. She was asymptomatic with a right upper abdominal mass that was smooth, mobile, and non-tender. Hepatitis virus markers and tumor markers were normal. The computed tomography (CT) images showed a 4.7×4.0 cm lesion in the left liver lobe with indistinct borders. Further magnetic resonance imaging (MRI) revealed low T1 and high T2 signal intensity with ring-shaped enhancement following contrast administration. Surgical resection was performed, and histology confirmed hepatic angioleiomyoma with thick-walled vessels and spindle cell proliferation. Immunohistochemistry was positive for SMA, desmin, caldesmon, CD31, and CD34. The patient had no recurrence during 5 years follow-up.

Comparative performance of electronegative membrane filtration and automated concentrating pipette for detection of antibiotic resistance genes and microbial markers in river water samples

The target viral and bacterial concentrations in river water are essential for environmental monitoring and public health studies. Filtration-based methods are commonly employed, yet challenges arise due to recoverability and filter pore size. This study aimed to compare the performance of electronegative membrane filtration (EMF) and automated Concentrating Pipette (CP) Select (InnovaPrep) methods for quantifying antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and bacterial and viral markers in river water samples. Fifty-four river water samples were collected from upstream and downstream locations in a river in Japan. The CP Select method was modified by adding MgCl2 and using different tips. The recovery efficiencies for total coliforms and Escherichia coli were assessed, and class 1 integron-integrase gene (intI1), 16S rRNA, gene encoding sulfonamide resistance (sul1), cross-assembly phage (crAssphage), pepper mild mottle virus (PMMoV), and Escherichia coli gene (sfmD) were detected. CP Select showed recovery efficiencies of 45 %–63 % for total coliforms and 17 %–35 % for E. coli. The intI1, 16S rRNA, sul1, crAssphage, PMMoV, and sfmD concentrations using the modified CP Select method were 10.1 ± 0.5, 8.7 ± 0.2, 7.7 ± 0.2, 6.7 ± 0.2, 5.4 ± 0.2, and 3.5 ± 0.5 log10 copies/L, respectively. Higher intI1 and sul1 concentrations were observed downstream, with the highest contribution percentage (22 % and 21 %) using CP Select or EMF. The modified CP Select method with 0.05 μm tips yielded more quantifiable results for all target genes and greater PMMoV concentrations (p < 0.05). Positive correlations were found among bacterial, ARG/MGE, and viral markers (Spearman's ρ = 0.71 for 16S rRNA and sfmD, 0.88 for intI1 and sul1, and 0.64 for PMMoV and crAssphage). The modified CP Select method demonstrated effective recovery of bacteria and quantification of ARGs, MGEs, and microbial markers in river water. Further studies are required to validate these methods and confirm their applicability in diverse environmental contexts.

Seroprevalence of hepatitis B virus markers of infection and immunity among people living in Libreville, Gabon

ObjectivesIn Gabon, data on hepatitis B virus (HBV) infection are limited to hepatitis B surface antigen (HBsAg) detection among specific populations and rural regions. This is the first study aimed at determining the seroprevalence of HBV markers among the Gabonese population. MethodsA retrospective study was conducted from January 2002 through December 2022. All patients who requested HBV marker screening (HBsAg, anti-HBc, anti-HBs, anti-HBe, hepatitis B e antigen) were included in this study. ResultsA total of 496 people were included in this study. The prevalence of HBV exposure was high (59.5%) and significantly associated with age, sex, pregnancy status, and social status. Among individuals with evidence of HBV exposure, only 89 (33.7%) had effectively cleared the virus. Overall, HBsAg was detected in 108 of the 496 (21.8%) people included in the study. HBsAg infection was more common among individuals in the 16-25 (35.1%) and 26-35-year (30.6%) age groups. Moreover, most of HBsAg-infected patients were health care workers (HCWs) (P = 0.0096). Among the 496 individuals included in this study, 126 (25.4%) were still susceptible to HBV. Most of the susceptible female patients were pregnant (P <0.0001). HCWs had a significantly lower probability of being naïve to HBV infection (P = 0.0001). There was a similar prevalence of susceptibility in male and female patients (50 [24.5%] male and 76 [26.0%] female patients). Only 15.1% of the studied population was vaccinated. ConclusionsOur findings revealed a high seroprevalence of HBV infection in the study area and very low HBV immunization coverage. Additional care and resources should be provided to promote HBV vaccination and block vertical HBV transmission.

Early biological markers of post-acute sequelae of SARS-CoV-2 infection

To understand the roles of acute-phase viral dynamics and host immune responses in post-acute sequelae of SARS-CoV-2 infection (PASC), we enrolled 136 participants within 5 days of their first positive SARS-CoV-2 real-time PCR test. Participants self-collected up to 21 nasal specimens within the first 28 days post-symptom onset; interviewer-administered questionnaires and blood samples were collected at enrollment, days 9, 14, 21, 28, and month 4 and 8 post-symptom onset. Defining PASC as the presence of any COVID-associated symptom at their 4-month visit, we compared viral markers (quantity and duration of nasal viral RNA load, infectious viral load, and plasma N-antigen level) and host immune markers (IL-6, IL-10, TNF-α, IFN-α, IFN-γ, MCP, IP-10, and Spike IgG) over the acute period. Compared to those who fully recovered, those reporting PASC demonstrated significantly higher maximum levels of SARS-CoV-2 RNA and N-antigen, burden of RNA and infectious viral shedding, and lower Spike-specific IgG levels within 9 days post-illness onset. No significant differences were identified among a panel of host immune markers. Our results suggest early viral dynamics and the associated host immune responses play a role in the pathogenesis of PASC, highlighting the importance of understanding early biological markers in the natural history of PASC.

Opisthotonic posturing in Guillian-Barre syndrome.

Here we report a child of Gullian Barre syndrome (GBS) with opisthotonic posturing and we subsequently detected Scrub typhus in him. An 11-year-old boy presented with progressive motor quadriparesis with transient bladder retention, bilateral facial weakness, diminished gag reflex, absent reflexes and his nerve conduction studies suggested Acute Motor Axonal Neuropathy (AMAN) GBS. His power gradually started recovering after one week. However, he had opisthotonus and signs of meningeal irritation. The child's CSF examination was consistent with GBS. His bacterial, fungal, tubercular microscopy and cultures and viral markers were negative. IgM for Borrelia and Leptospira and HIV ELISA were negative. IgM for scrub typhus, however, came out to be positive. Hence, we gave him azithromycin and he recovered almost completely in 3 months. To our knowledge there is no previous report of opisthotonic posturing in GBS patients. This could be due to radicular involvement in immune mediated etiology of GBS.

Comprehensive Clinical Profile and Hemodialysis Outcomes in Patients Attending a Tertiary Care Hospital.

Background Chronic kidney disease (CKD) can lead to serious conditions such as anemia and cardiovascular disease, posing a growing global health challenge. End-stage renal disease (ESRD) requires treatments such as dialysis or kidney transplantation. Despite the widespread impact and rising prevalence of CKD and ESRD, comprehensive data remains limited in India. This study seeks to investigate the clinical, socio-demographic, and etiological profiles of CKD patients undergoing hemodialysis at a tertiary care hospital, with the goal of enhancing understanding and improving patient care. Methodology This retrospective cohort study, conducted at a tertiary care center, included 500 CKD patients undergoing hemodialysis, with comprehensive medical records. Data collected covered demographics (age, sex, education, and occupation), CKD etiology, disease duration, hemodialysis duration, viral marker status, blood transfusions, and vascular access details. With continuous variables reported as mean ± standard deviation (SD) and categorical variables as counts (percentages), statistical analysis was carried out using SPSS version 21 (IBM Corp., Armonk, New York, USA). The connections were examined using the Pearson Chi-square test, with P≤0.05 being deemed significant. Results The study revealed that hypertension was the primary cause of CKD in 58% of patients, followed by diabetes mellitus in 13%. A significant 93% of patients tested negative for viral markers such as human immunodeficiency virus (HIV), hepatitis C virus (HCV), and hepatitis B surface antigen (HBsAg). Hemodialysis duration varied, with 68% of patients undergoing dialysis for one to five years. Most patients had two (40%) or three (58%) dialysis sessions per week, and 84% had only one arteriovenous (AV) fistula surgery. Blood transfusions were common, with 62% of patients receiving between one and five transfusions. The gender distribution showed more males (372) than females (201), and the majority of patients were aged between 41 and 60 years. Conclusion This study highlights the importance of early detection and management of CKD, emphasizing preventive health measures, enhanced diagnostic capabilities, and sufficient resource allocation to reduce the disease burden. It also calls for further research into unknown CKD causes and strategies to improve patient care and outcomes.