High Viral Load Research Articles

Comparative study of two Rift Valley fever virus field strains originating from Mauritania.

Rift Valley fever (RVF) is one of the major viral arthropod-borne diseases in Africa. In recent decades, RVF virus (RVFV), the causative agent of RVF, has been responsible for multiple outbreaks in West Africa with important consequences on human and animal health. In particular, an outbreak occurred in 2010 after heavy rains in the desertic region of Adrar, Mauritania. It was characterized by the appearance of severe clinical signs among dromedary camels. Another one occurred in 2013-2014 across Senegal and the southern part of Mauritania. In this study, we characterized two RVFV field strains isolated during these two outbreaks. The first strain, MRU25010-30, was isolated from a camel (2010) while the second, MRU2687-3, was isolated from a goat (2013). By deep-sequencing and rapid amplification of cDNA-ends by polymerase chain reaction, we successfully sequenced the complete genome of these two RVFV strains as well as the reference laboratory strain ZH548. Phylogenetic analysis showed that the two field viruses belong to two different RVFV genetic lineages. Moreover, we showed that MRU25010-30 replicates more efficiently in various in vitro cell culture models than MRU2687-3 and ZH548. In vivo, MRU25010-30 caused rapid death of BALB/c mice and proved to be more virulent than MRU2687-3, regardless of the route of inoculation (subcutaneous or intranasal). The virulence of MRU25010-30 is associated with a high viral load in the liver and serum of infected mice, while the death of mice infected with MRU2687-3 and ZH548 correlated with a high viral load in the brain. Altogether, the data presented in this study provide new avenues to unveil the molecular viral determinants that modulate RVFV virulence and replication capacity.

Optimal thresholds and key parameters for predicting influenza A virus transmission events in ferrets

Although assessments of influenza A virus transmissibility in the ferret model play a critical role in pandemic risk evaluations, few studies have investigated which virological data collected from virus-inoculated animals are most predictive of subsequent virus transmission to naïve contacts. We compiled viral titer data from >475 ferrets inoculated with 97 contemporary IAV (including high- and low-pathogenicity avian, swine-origin, and human viruses of multiple HA subtypes) that served as donors for assessments of virus transmission in the presence of direct contact (DCT) or via respiratory droplets (RDT). A diversity of molecular determinants, clinical parameters, and infectious titer measurements and derived quantities were examined to identify which metrics were most statistically supported with transmission outcome. Higher viral loads in nasal wash (NW) specimens were strongly associated with higher transmission frequencies in DCT, but not RDT models. However, viruses that reached peak titers in NW specimens early (day 1 p.i.) were strongly associated with higher transmission in both models. Interestingly, viruses with ‘intermediate’ transmission outcomes (33–66%) had NW titers and derived quantities more similar to non-transmissible viruses (<33%) in a DCT setting, but with efficiently transmissible viruses (>67%) in a RDT setting. Machine learning was employed to further assess the predictive role of summary measures and varied interpretation of intermediate transmission outcomes in both DCT and RDT models, with models employing these different thresholds yielding high performance metrics against both internal and external datasets. Collectively, these findings suggest that higher viral load in inoculated animals can be predictive of DCT outcomes, whereas the timing of when peak titers are detected in inoculated animals can inform RDT outcomes. Identification that intermediate transmission outcomes should be contextualized relative to the transmission mode assessed provides needed refinement towards improving interpretation of ferret transmission studies in the context of pandemic risk assessment.

Effect of HIV and Malaria in Pregnancy on Pertussis-specific Antibodies and Transplacental Antibody Transfer: A Secondary Analysis of a Prospective Cohort Study in Mozambican Pregnant Women and Their Infants.

Infection during pregnancy may affect maternal and infant immunity against childhood diseases. We aimed to evaluate the effects of maternal HIV and malaria on maternal and infant pertussis immunity and placental antibody transfer. A prospective study was conducted in mother-infant pairs in Mozambique. Peripheral and cord blood samples were collected for pertussis-specific immune assays. Maternal HIV serostatus and Plasmodium falciparum infection were assessed. The placental transfer was assessed using cord-to-mother ratios of IgG against pertussis toxin (PT), pertactin (PRN) and fimbriae 2/3 (FIM). A total of 270 mother-infant pairs were included: 99 mothers with HIV and 40 mothers with malaria. Pregnant women with HIV showed a reduction in placental transfer [PT: 12.7%, 95% confidence interval (CI): 2.6-21.7, P = 0.015; PRN: 14.6%, 95% CI: 6.3-22.1, P = 0.001; and FIM: 7.5%, 95% CI: -6.6 to 19.7, P = 0.282] compared with women without HIV. A trend toward reduction in IgG transfer was observed among women with malaria (PT: 9.5%, 95% CI: -4.2 to 21.4, P = 0.165; PRN: 5.0%, 95% CI: -7.0 to 15.7, P = 0.394; and FIM: 15.9%, 95% CI: -0.9 to 30.0, P = 0.062) compared with those without. Maternal HIV infection (odds ratio: 4.43, 95% CI: 2.14-9.1; P < 0.001) and high viral load (odds ratio: 4.37, 95% CI: 1.4-12.2; P = 0.033) were associated with impaired placental transfer. Maternal HIV infection is associated with lower mother-to-infant transfer of pertussis antibodies. While efforts continue in the health care of pregnant women with HIV, interventions such as maternal immunization can be a valuable strategy to prevent pertussis in infants.

Human adenovirus type 7 (HAdV-7) infection induces pulmonary vascular endothelial injury through the activation of endothelial autophagy

BackgroundHAdV-7 is a prevalent pathogen that can cause severe pneumonia in children. Previous studies have shown a significant increase in serum levels of vascular permeability factor (VPF/VEGF) and viral load in pediatric patients with fatal HAdV-7 infection, suggesting potential damage to the pulmonary vascular endothelium. Further research is necessary to elucidate the underlying mechanism.MethodsThe human lung microvascular endothelial cell line-5a and human CD46 mice were used for in vitro and in vivo experiments, respectively. RNA-seq was employed for correlative omics analysis. Viral infection and copy status were examined using transmission electron microscopy to observe virus particles, immunofluorescence to detect the viral protein Hexon, and qPCR to assess HAdV-7 fiber gene copies. Various methods, including ELISAs for VEGF and other injury markers, the CCK8 assay for cell viability, and flow cytometry for endothelium numbers, were employed to evaluate endothelial damage. Acute lung injury severity was evaluated by scoring pathological inflammation and measuring pulmonary vascular permeability. Autophagy activation was assessed by observing autophagosomes and validating marker proteins.ResultsGSEA analysis showed significant enrichment of gene sets related to endothelial functions (barrier, defense, and regeneration) and ALI in the HAdV-7-infected group. GO analysis indicated an enrichment of autophagy-related pathways linked to cell death. Subsequently, successful signs of HAdV-7 infection and replication were observed in the endothelium, including cytopathic effects, intracellular virions, and increased HAdV-7 fiber gene copies. Endothelial injury, including mitochondrial damage, decreased endothelium, and elevated levels of endothelial injury markers such as VEGF, sICAM-1, sVCAM-1, E-selectin, ESM1, MCP1, and IL1β were observed after HAdV-7 infection. Additionally, evidence of leaky lung blood vessels and ALI was observed, including progressive weight loss, elevated pulmonary vascular permeability, and severe lung consolidation. Furthermore, HAdV-7 infection induced autophagosome formation in the endothelium and triggered complete cell autophagy. Importantly, inhibiting autophagic flux reduced VEGF levels and other endothelial injury markers, decreased viral load, improved cell survival rate, alleviated pulmonary vessel leakage, and mitigated lung inflammation.ConclusionsHAdV-7 successfully infects pulmonary vascular endothelium and replicates effectively, causing injury to the endothelium, high VEGF expression and viral load in the serum, as well as ALI/ARDS. Autophagy inhibitors can alleviate endothelial injury, inhibit viral replication, relieve leakage from the vasculature, and reduce lung inflammation.

The Co-Infection of ISKNV-II and RGNNV Resulting in Mass Mortality of Juvenile Asian Seabass (Lates calcarifer), Zhuhai, Southern China.

Infectious spleen and kidney necrosis virus (ISKNV) and nervous necrosis virus (NNV) are two common and important causative agents in marine-cultured fish. However, high viral loads of both ISKNV and NNV in the same clinical case is unusual. In this study, a mass mortality event of Asian seabass Lates calcarifer juveniles occurred in Zhuhai, the main Asian seabass cultured area in mainland China. The fish samples were pooled for pathogen identification and both high viral loads of ISKNV and NNV were detected by real-time microfluidic quantitative PCR and conventional PCR. Immunohistochemistry and immunofluorescence showed that strong ISKNV signals were detected in spleen and liver, while strong NNV signals were detected in brain and eye. The tissue homogenates were inoculated into MFF-1 cell and SSN-1 cell, respectively. After several viral passages, both individual ISKNV and NNV were purely isolated from each other, and designated as ASB-ISKNV-23 and ASB-NNV-23, respectively. The whole genome sequences of ASB-ISKNV-23 and ASB-NNV-23 were determined and annotated. The result showed that ASB-ISKNV-23 and ASB-NNV-23 are composed of 112,236 bp and 4441 bp, respectively. Phylogeny analysis showed that ASB-ISKNV-23 belongs to ISKNV-II sub-genotype and ASB-NNV-23 belongs to RGNNV genotype. Collectedly, coinfection of ISKNV-II and RGNNV were firstly documented in mass mortality of Asian seabass in mainland China. Importantly, both individual ISKNV-II and RGNNV were purely isolated using two different permissive cell lines. Our study provides useful information for better understanding the complex pathogenesis regarding the coinfection with ISKNV and NNV in farmed fish.

Study on the Clinical Characteristics, Persistent Infection Ability and Viral Load of Human Papillomavirus Type 73 Single Infection.

Human papillomavirus (HPV) infection is a main cause of cervical cancer. Different HPV types exhibit significant differences in pathogenicity. Clinical data on HPV73 single infection is limited, this study aimed to analyze the clinical characteristics of HPV73 single infections. We collected cervical exfoliated cells to examine the infection rate and the clinical characteristics of patients with single HPV73 infection, and to investigate the association of viral load, persistent infection, and pathogenicity. The positive rate of HPV73 among women tested was approximately 0.14% (609/435 072). Among 214 HPV73 single infection cases, the number of patients with normal cytology, cervical intraepithelial neoplasia (CIN)1, CIN2, CIN3, and cervical cancer were 182, 24, 5, 1, and 1, respectively. Follow-up data showed that 14.29% (3/21) of CIN1 patients developed to higher-grade lesions, with 3.73% (5/134) of normal patients progressed to CIN. The viral load in the CIN group was significantly higher than that in the normal group (p = 0.001), and the viral load in the persistent infection group was significantly higher than that of the viral-clearance group (p < 0.001). HPV73 single infection can lead to cervical cancer, with relatively low pathogenicity. And high viral load is a crucial factor in its persistent infection and pathogenicity.

Rhabdomyolysis in Severe Fever With Thrombocytopenia Syndrome: Associations With Acute Kidney Injury and Mortality.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging zoonosis caused by a novel bunyavirus. Until recently, rhabdomyolysis in SFTS was not elucidated. The objective of this study was to investigate the prevalence and clinical characteristics of rhabdomyolysis and its association with AKI and prognosis in patients with SFTS. A total of 231 consecutive patients diagnosed with SFTS were enrolled, including 144 (62.3%) patients in the no rhabdomyolysis group and 87 (37.7%) patients in the rhabdomyolysis group. Clinical characteristics and laboratory parameters of SFTS patients were compared between the no rhabdomyolysis and rhabdomyolysis groups. Patients with rhabdomyolysis had more frequency of confusion, cough, sputum, chest distress, abdominal pain, petechia and ecchymosis, headache, myalgia, and myasthenia than those without rhabdomyolysis. Compared with patients in the no rhabdomyolysis group, patients in the rhabdomyolysis group had higher serum levels of laboratory parameters referring to liver, kidney, pancreas, heart, coagulation system injury, and higher viral load. The cumulative survival rate of patients with rhabdomyolysis was significantly lower than that of patients without rhabdomyolysis. Furthermore, univariate and multivariate binary logistic regression analyses demonstrated that rhabdomyolysis was an independent predictor for acute kidney injury and mortality in patients with SFTS. Rhabdomyolysis may be an important contributing factor to adverse outcomes and its effects on mortality may be mediated by acute kidney injury in SFTS. The early detection and effective intervention of rhabdomyolysis may decrease the mortality of patients with SFTS.

A prospective, multicenter study of hepatitis B birth-dose vaccine with or without hepatitis B immunoglobulin in preventing mother-to-child transmission of hepatitis B virus in Ethiopia

BackgroundHistorically, mother-to-child transmission (MTCT) of hepatitis B virus (HBV) was considered uncommon in Africa, leading to a reluctant attitude to birth-dose HBV vaccination on the continent. As a randomized trial would be unethical, real-life data are needed to assess the effect of HBV birth-dose vaccine in Africa. MethodsA multicenter, prospective, observational study of hepatitis B surface antigen (HBsAg)-positive pregnant women and their infants was carried out in Ethiopia, from January 2019 to May 2021. Pregnant women were screened for HBsAg and HIV as part of routine antenatal care and/or delivery, and HBsAg-positive HIV-negative pregnant women were included in the study. HBV birth-dose vaccine and hepatitis B immunoglobulin (HBIg) were recommended but not all newborns received it as it was not national policy. All infants, however, received the pentavalent HBV vaccine at 6, 10, and 14 weeks of age. Vaccination status was confirmed from delivery ward charts and infant vaccination certificates. Infants were tested for HBsAg at 9 months of age and a positive result was taken as evidence of MTCT. FindingsOf 290 HBsAg-positive pregnant women, 168 mother/infant pairs returned for their 9-month follow-up visit and were included in this analysis. Two of 112 (1.8 %) infants who received birth-dose vaccine with HBIg, and 2 of 23 (8.7 %) who received birth-dose vaccine alone were HBsAg positive at nine months of age, compared to 8 of 33 (24.2 %) who received neither vaccine nor HBIg at birth (p = 0.002). High maternal viral load (>200,000 IU/ml; adjusted odds ratio [AOR] 10.4; 95 % confidence interval [CI] 1.2–92.1) and not receiving HBV birth-dose vaccine nor HBIg (AOR 29.2; 95 % CI 4.0–211.3) were independent predictors of MTCT. InterpretationBirth-dose HBV vaccine with or without HBIg significantly reduced the risk of MTCT of HBV in Ethiopia. Improved coverage of birth-dose HBV vaccine should be an urgent priority.

Evaluating compliance with quality indicators of HIV and AIDS clinical care and analyzing those associated with mortality in the Democratic Republic of Congo

BackgroundSince human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) infection have been identified, significant improvements have been made concerning its diagnosis and treatment. Few contributions have been made in the area of quality indicators in the Democratic Republic of Congo (DRC). To address this gap, a recent study conducted in the said area in 2023 resulted in the development of a comprehensive list of 88 indicators for assessing the quality of clinical care for HIV and AIDS. Out of these, 66 were identified as a core set of quality indicators for the assessment of HIV and AIDS clinical care. The aim of this study was to evaluate compliance with these quality indicators and determine those that are associated with mortality among people living with HIV and AIDS (PLWHA) in the DRC.MethodsA retrospective cohort study was conducted at Panzi Hospital in Bukavu, South Kivu province of the DRC. The study population included PLWHA adult patients who were followed for a period of five years at the antiretroviral (ARV) clinic from January 2017 to December 2021. Descriptive analysis was performed to evaluate essential characteristics, and inferential statistics were performed including a multivariable logistic regression analysis performed to assess the association between selected clinical care indicators and PLWHA mortality, to estimate the odds ratios of mortality based on various factors, and to control for confounding variables.ResultsA total of 884 PLWHA were included. The level of compliance with the quality indicators was 78.79%. In the multivariate analysis, the quality-of-care indicators associated with mortality in the follow-up PLWHA on treatment were items 12 (Late diagnosis of HIV infection in specialized care), 24 (Assessment of resistance in cases of virologic failure), and 38 (Evaluation of Cardiac risk). Other factors associated with mortality were male sex, lower CD4 (< 500mm3) and high viral load (> 50 copies/ml).ConclusionThe level of compliance with the indicators was moderate. Some quality indicators were associated with mortality. These indicators are associated with delayed diagnosis of HIV infection, assessment of resistances in case of virologic failure, and evaluation of cardiac risk. Addressing gaps in adherence to these quality of care indicators (clinical care standards) could potentially reduce mortality rates and enhance the overall management of HIV and AIDS, particularly at Panzi Hospital in Bukavu, South Kivu Province, and in general in the DRC.

Molecular analysis of the 2022 mpox outbreak and antiviral activity of dihydroorotate dehydrogenase inhibitors against orthopoxviruses

Monkeypox virus (MPXV) has caused a large pandemic outbreak in 2022 with more than 90.000 confirmed cases and 181 deaths. Notably, signs of microevolution and host adaption have been observed. Here, we demonstrate that viral genomes from Franconia, Bavaria acquired different mutations. Three isolates obtained from diagnostic samples, submitted from suspected Mpox cases, show differences in their replication capacities. One MPXV isolate which shows the fastest replication kinetics and higher viral loads, possesses a unique non-synonymous mutation (D616L) in the A11L protein (gene OPG136), which encodes for a protein that is part of a major viral core structure. In regard to pandemic preparedness and future outbreaks, we analyzed the antiviral activity of dihydroorotate dehydrogenase (DHODH) inhibitors, and show that they are active against MPXV, vaccinia virus (VACV), and cowpox virus (CPXV) and therefore likely against orthopoxviruses in general. In agreement with that, we also demonstrated that chemical optimization leads to compounds with EC50 values in the sub-nanomolar range, associated with low cytotoxicity, which forms a good basis for future drug development from this chemical series.

Rapid test for hepatitis B core-related antigen to identify people living with hepatitis B having high viral load in Cameroon

This study presents a retrospective assessment of the diagnostic performance of the newly developed hepatitis B core-related antigen rapid diagnostic test (HBcrAg-RDT) in detecting plasma samples with elevated hepatitis B virus (HBV) DNA levels (≥200,000 IU/ml) in Yaoundé, Cameroon. Samples were collected consecutively from treatment-naïve adults living with HBV between January 1, 2021, and June 30, 2023. Analyzing 146 samples from participants with a median age of 36 years, the HBcrAg-RDT exhibited a sensitivity of 97.5% (95% CI: 86.8–99.9) and a specificity of 77.4% (68.2–84.9) when compared to real-time PCR as the reference standard. These findings suggest that HBcrAg-RDT holds promise as a valuable point-of-care tool for diagnosing high HBV DNA levels, particularly in resource-limited settings. Further research will refine its practicality and effectiveness.

Quantification of hepatitis B viral load in patients attending healthcare facilities in Gezira state, Sudan

Abstract Background Hepatitis B virus (HBV) infection remains a significant global health challenge particularly in developing countries and regions. Gezira State, Sudan where prevalence and disease burden are high is an example of such regions. This study aimed to quantify HBV viral load among patients in Gezira State and assess the distribution of viral loads across demographic groups, including gender and age. Method A descriptive cross-sectional study was conducted between January 2022 and January 2023. Randomly selected 100 chronic HBV patients were enrolled from major healthcare facilities in Wad Medani, Gezira’s capital. Viral loads were measured using real-time PCR, and statistical analyses were performed to identify predictors of high viral load. Results The results revealed that the mean HBV viral load was 886,363 IU/ml (SD± 421,295), with a significantly higher mean in males (1,122,899 IU/ml) compared to females (531,559 IU/ml) (p&lt; 0.0001). Viral loads ranged from 15 IU/ml to 35,474,488 IU/ml, with 37% of patients exceeding the WHO-recommended treatment threshold of 2,000 IU/ml. However, no significant association was found between viral load and age (p=0.939). Conclusion The study highlighted the need for gender-specific management strategies and the importance of HBV viral load quantification for effective public health intervention in Gezira State. Further research is recommended to explore HBV epidemiology in Sudan more comprehensively.

Leveraging environmental microbial indicators in wastewater for data-driven disease diagnostics.

Wastewater-based surveillance (WBS) is an emerging tool for monitoring the spread of infectious diseases, such as SARS-CoV-2, in community settings. Environmental factors, including water quality parameters and seasonal variations, may influence the prevalence of viral particles in wastewater. This study aims to explore the relationships between these factors and the incidence of SARS-CoV-2 across 28 monitoring sites, spanning different seasons and water strata. Samples were collected from 28 sites, accounting for seasonal and spatial (surface and intermediate water layers) variations. Key physicochemical parameters, heavy metals, and minerals were measured, and viral presence was detected using RT-qPCR. After data preprocessing, correlation analyses identified 19 relevant environmental parameters. Unsupervised learning algorithms, including K-means and K-medoid clustering, were employed to categorize the data into four distinct clusters, revealing patterns of viral positivity and environmental conditions. Cluster analysis indicated that seasonal variations and water quality characteristics significantly influenced SARS-CoV-2 positivity rates. The four clusters demonstrated distinct associations between environmental factors and viral prevalence, with certain clusters correlating with higher viral loads in specific seasons. The clustering patterns varied across sample sites, reflecting the diverse environmental conditions and their influence on viral detection. The findings underscore the critical role of environmental factors, such as water quality and seasonality, in shaping the dynamics of SARS-CoV-2 prevalence in wastewater. These insights provide a deeper understanding of the complex interplay between environmental contexts and disease spread. By utilizing WBS and advanced data analysis techniques, this study offers a robust framework for future research aimed at enhancing public health surveillance and interventions.

Study on the clinical characteristics, persistent infection capability and viral load of human papillomavirus type 26 single infection

BackgroundHuman papillomavirus (HPV) infection is the main cause of cervical cancer. Different types of HPV have varying carcinogenic capabilities, and viral load is one of the key indicators of pathogenicity. Currently, there is a lack of clinical data on HPV26. This study analyzed the clinical characteristics of patients with HPV26 single infection.MethodsExfoliated cervical cells were collected for HPV genotyping from women who attended gynecological outpatient clinics or physical examinations. The clinical characteristics of HPV26 single infections in both cross-sectional and follow-up studies were examined, and the association of viral load with HPV26 persistent infection and pathogenicity was investigated.ResultsThe HPV26 positive rate among women visiting hospitals for gynecological medical consultation or physical examination was approximately 0.09% (379/435,072). Among the HPV types tested, the detection rate of HPV26 was 0.37% (379/103,608). In the cross-sectional histopathological study of 101 patients with HPV26 single infection, 62.37% (63/101) presented lesion-free. The numbers of patients with cervical intraepithelial neoplasia (CIN) 1, CIN2, and CIN3 were 25, eight, and five, respectively. Cervical cancer was not detected in any patient. 71 patients attended follow-up examinations as well as HPV26 retesting up to two years, during which, 28.57% (6/21) of CIN1 patients have developed into high-grade lesions, and 9.61% (5/52) of lesion-free patients have progressed to CIN stage. The viral load in the CIN group was significantly higher than that in the lesion-free group (p = 0.012). Similarly, the viral load in the persistent infection group was significantly higher than that in the viral-clearance group (p < 0.001).ConclusionsThe pathogenicity of single HPV26 infections is moderate among high-risk types, warranting the inclusion of HPV26 in expanded screening for HPV. High viral load is an important factor in the persistent infection and pathogenicity of HPV26. Viral load is expected to serve as a screening risk factor for persistent infection and disease progression associated with HPV26.

VRK2 inhibits the replication of infectious bursal disease virus by phosphorylating RACK1 and suppressing apoptosis

Infectious bursal disease (IBD) is an acute, highly contagious, and immunosuppressive avian disease caused by the infectious bursal disease virus (IBDV). Despite significant efforts, the lack of knowledge about host proteins that counteract IBDV replication has hindered progress in preventing and controlling IBD in chickens. This study identifies the mitochondria-associated protein vaccinia virus-related kinase 2 (VRK2) as an inhibitor of IBDV. Overexpression of VRK2 significantly reduced IBDV proliferation in DF-1 cells and chicken embryo fibroblasts (CEFs). Conversely, the absence of VRK2 resulted in higher viral loads in these cells. Additionally, we found that VRK2 interacts with voltage-dependent anion channel 2 (VDAC2) and Receptor for Activated C Kinase 1 (RACK1). Mechanistic studies revealed that VRK2 inhibits IBDV-induced apoptosis by targeting RACK1 phosphorylation, leading to reduced viral growth. This study enhances our understanding of VRK2's role in host anti-apoptotic mechanisms and offers novel insights into IBDV pathogenesis and vaccine development.

The effect of the modifiable areal unit problem (MAUP) on spatial aggregation of COVID-19 wastewater surveillance data

Large wastewater-based epidemiology (WBE) projects often have wide coverage and multiple sampling sites, necessitating spatial aggregation for data reporting and interpretation. However, the outcome may be impacted by a type of statistical bias called the modifiable areal unit problem (MAUP). In this study, we examined the presence and extent of the MAUP scaling effect on a New York State COVID-19 wastewater surveillance project. Specifically, we investigated three metrics: 1) the difference in wastewater SARS-CoV-2 concentrations between sampling at city-level site (i.e., city's primary wastewater treatment plant influent stream) and at upstream sampling sites; 2) the correlation between WBE data and clinical indicators at the WWTP-level and the more aggregated county-level; and 3) the proportion of population affected by misalignment of COVID-19 community risk levels at different spatial scales. The results showed that the MAUP can have a negative impact on risk perception by masking regions with high wastewater viral load or COVID-19 community risk level. On the other hand, the MAUP improved the correlation between wastewater surveillance and clinical measures by an average of 26.02 %. This is the first study to investigate the MAUP in the context of WBE and may encourage future WBE projects to consider the implications of the MAUP when interpreting and reporting spatial data, ultimately leading to better data representativeness and accuracy.

Predicting Immune Flares in Untreated Chronic Hepatitis B Patients Using Novel Risk Factors and the FLARE-B Score.

Risk factors of chronic hepatitis B (CHB) immune flares are poorly understood. The primary aim of this study was to discover predictors of the CHB flare in non-cirrhotic, untreated CHB patients and develop a simple risk-stratifying score to predict the CHB flare. The secondary aim was to compare different machine learning methods for prediction. A retrospective cohort of untreated, non-cirrhotic CHB patients with normal baseline ALT was followed up over time until an immune flare as defined by ALT twice the upper limit of normal. Statistical learning and machine learning algorithms were used to develop predictive models using baseline variables. Bootstrap validation was used to internally validate the models. Of 405 patients (median age 44y; 41% male, 10% HBeAg positive), 67 (17%) experienced an immune flare by 5 years (annual incidence 4.0%). Predictors of flare included raised serum globulin, younger age, HBeAg positive status, higher viral load and raised liver stiffness. A simple predictive model "FLARE-B" had optimism-adjusted 1, 3 and 5-year AUCs of 0.813, 0.728 and 0.702, respectively. The random survival forest algorithm had the highest optimism-adjusted AUCs of 0.861, 0.766 and 0.725, respectively. New, novel predictors of the CHB flare include a raised serum globulin and possibly raised liver stiffness and the absence of liver steatosis. FLARE-B can be used to risk-stratify individuals and potentially guide personalized management strategies such as monitoring schedules and proactive antiviral treatment in high-risk patients.

Factors associated with High Viral Loads among HIV Patients Under ARV at Gitwe District Hospital

Background: High viral load is a condition of rapid multiplication in blood stream. The high viral load among HIV patients under ARV may also occur due to the rapid multiplication of virus leading to compromise immune system of living organism up to death. The vast majority of people living with HIV virus face with persistent of high viral load while were under treatments (ARV). Aim: The aim of the study was to evaluate and assess the factors associated with high viral load among HIV patients under ARV at Gitwe District Hospital. Methodology: The retrospective study was conducted to assess high viral load of HIV patients under ARV. The only significance factor for viral load were adherence with (p=0.000173) and ages with (p=0.00023). Results and conclusion: A large of number of PLWH were highly VL tested, and 95 subjects were recorded for study. Among 95 subjects, 54.73% were female and 45.26% male. All data were analyzed using statistical package for social science (SPSS) version 20. The results have showed provider’s advices and ART use, adherence counselling. Keywords: High viral load, HIV, Antiretroviral therapy, Gitwe District Hospital.

Longitudinal 1H NMR-Based Metabolomics in Saliva Unveils Signatures of Transition from Acute to Post-Acute Phase of SARS-CoV-2 Infection.

COVID-19 can range from a mild to severe acute respiratory syndrome and also could result in multisystemic damage. Additionally, many people develop post-acute symptoms associated with immune and metabolic disturbances in response to viral infection, requiring longitudinal and multisystem studies to understand the complexity of COVID-19 pathophysiology. Here, we conducted a 1H Nuclear Magnetic Resonance metabolomics in saliva of symptomatic subjects presenting mild and moderate respiratory symptoms to investigate prospective changes in the metabolism induced after acute-phase SARS-CoV-2 infection. Saliva from 119 donors presenting non-COVID and COVID-19 respiratory symptoms were evaluated in the acute phase (T1) and the post-acute phase (T2). We found two clusters of metabolite fluctuation in the COVID-19 group. Cluster 1, metabolites such as glucose, (CH3)3 choline-related metabolites, 2-hydroxybutyrate, BCAA, and taurine increased in T2 relative to T1, and in cluster 2, acetate, creatine/creatinine, phenylalanine, histidine, and lysine decreased in T2 relative to T1. Metabolic fluctuations in the COVID-19 group were associated with overweight/obesity, vaccination status, higher viral load, and viral clearance of the respiratory tract. Our data unveil metabolic signatures associated with the transition to the post-acute phase of SARS-CoV-2 infection that may reflect tissue damage, inflammatory process, and activation of tissue repair cascade. Thus, they contribute to describing alterations in host metabolism that may be associated with prolonged symptoms of COVID-19.

Major alteration of lung microbiome and the host responses in critically ill COVID-19 patients with high viral load

Patients with COVID-19 under invasive mechanical ventilation are at higher risk of developing ventilator-associated pneumonia (VAP), associated with increased healthcare costs, and unfavorable prognosis. The underlying mechanisms of this phenomenon have not been thoroughly dissected. Therefore, this study attempted to bridge this gap by performing a lung microbiota analysis and evaluating the host immune responses that could drive the development of VAP. In this prospective cohort study, mechanically ventilated patients with confirmed SARS-CoV-2 infection were enrolled. Nasal swabs (NS), endotracheal aspirates (ETA), and blood samples were collected initially within 12 h of intubation and again at 72 h post-intubation. Plasma samples underwent cytokine and metabolomic analyses, while NS and ETA samples were sequenced for lung microbiome examination. The cohort was categorized based on the development of VAP. Data analysis was conducted using RStudio version 4.3.1. In a study of 36 COVID-19 patients on mechanical ventilation, significant differences were found in the nasal and pulmonary microbiome, notably in Staphylococcus and Enterobacteriaceae, linked to VAP. Patients with VAP showed a higher SARS-CoV-2 viral load in respiratory samples, elevated neutralizing antibodies, and reduced inflammatory cytokines, including IFN-δ, IL-1β, IL-12p70, IL-18, IL-6, TNF-α, and CCL4. Metabolomic analysis revealed changes in 22 metabolites in non-VAP patients and 27 in VAP patients, highlighting D-Maltose-Lactose, Histidinyl-Glycine, and various phosphatidylcholines, indicating a metabolic predisposition to VAP. This study reveals a critical link between respiratory microbiome alterations and ventilator-associated pneumonia in COVID-19 patients with higher SARS-CoV-2 viral loads in respiratory samples, elevated neutralizing antibodies, and reduced inflammatory cytokines, including IFN-δ, IL-1β, IL-12p70, IL-18, IL-6, TNF-α, and CCL4. These findings provide novel insights into the underlying mechanisms of VAP, with potential implications for management and prevention.