Application of wastewater-based surveillance and copula time-series model for COVID-19 forecasts.

The objective of this study was to develop a novel copula-based time series (CTS) model to forecast COVID-19 cases and trends based on wastewater SARS-CoV-2 viral load and clinical variables. Wastewater samples were collected from wastewater pumping stations in five sewersheds in the City of Chesapeake VA. Wastewater SARS-CoV-2 viral load was measured using reverse transcription droplet digital PCR (RT-ddPCR). The clinical dataset included daily COVID-19 reported cases, hospitalization cases, and death cases. The CTS model development included two steps: an autoregressive moving average (ARMA) model for time series analysis (step I), and an integration of ARMA and a copula function for marginal regression analysis (step II). Poisson and negative binomial marginal probability densities for copula functions were used to determine the forecasting capacity of the CTS model for COVID-19 forecasts in the same geographical area. The dynamic trends predicted by the CTS model were well suited to the trend of the reported cases as the forecasted cases from the CTS model fell within the 99 % confidence interval of the reported cases. Wastewater SARS CoV-2 viral load served as a reliable predictor for forecasting COVID-19 cases. The CTS model provided robust modeling to predict COVID-19 cases.

A Current View of SARS-CoV-2 Vaccine Policies Amongst Pediatric Renal Transplant Centers and Relationships to State Policy.

Given its mortality benefit, renal transplantation remains the ideal treatment modality for end stage renal disease in children. Despite the recent expansion of use in young children, the novel SARS-CoV-2 vaccine has not been universally accepted. Similarly, vaccine related state regulations are heterogenous. We present a cross-sectional analysis of institutional specific vaccination policies at US pediatric renal transplant centers and relationships to state legislation. We found that 36.1% of institutions require COVID-19 vaccination prior to transplant, while 17 states have current legislation prohibiting proof of vaccination as a means of access to public services. Of the 63.9% of transplant centers without immunization requirement, almost two-thirds are located in states without prohibitory regulations. Despite an unclear primary influence of institutional policy, our study demonstrates a lack of standardization and potential to create unnecessary inequities.

Collection system investigation microbial source tracking (CSI-MST): Applying molecular markers to identify sewer infrastructure failures.

Collection System Investigation Microbial Source Tracking (CSI-MST) is a novel, sensitive approach for identifying sewer infrastructure deficiencies using molecular markers. This method requires both a detailed understanding of collection and conveyance system infrastructure and quickly turned around molecular data to advise an adaptive, targeted in-pipe approach to detect deficiencies. Here we explain the CSI-MST approach and provide several case study examples of how this approach can be adapted to different scale watersheds to identify potential sewer infrastructure issues. This approach has been used to locate and confirm the remediation of numerous needed infrastructure repairs in the southeastern Virginia region. The selected case studies presented here serve as a proof of concept-this methodology can be adopted by other utilities and municipalities to address necessary wastewater infrastructure repairs in different regions.

COVID-19 surveillance in Southeastern Virginia using wastewater-based epidemiology.

Wastewater-based epidemiology (WBE) has been used to analyze markers in wastewater treatment plant (WWTP) influent to characterize emerging chemicals, drug use patterns, or disease spread within communities. This approach can be particularly helpful in understanding outbreaks of disease like the novel Coronavirus disease-19 (COVID-19) when combined with clinical datasets. In this study, three RT-ddPCR assays (N1, N2, N3) were used to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in weekly samples from nine WWTPs in southeastern Virginia. In the first several weeks of sampling, SARS-CoV-2 detections were sporadic. Frequency of detections and overall concentrations of RNA within samples increased from mid March into late July. During the twenty-one week study, SARS-CoV-2 concentrations ranged from 101 to 104 copies 100 mL-1 in samples where viral RNA was detected. Fluctuations in population normalized loading rates in several of the WWTP service areas agreed with known outbreaks during the study. Here we propose several ways that data can be presented spatially and temporally to be of greatest use to public health officials. As the COVID-19 pandemic wanes, it is likely that communities will see increased incidence of small, localized outbreaks. In these instances, WBE could be used as a pre-screening tool to better target clinical testing needs in communities with limited resources.

Integrating Bayesian Analysis and Cumulative Probability Generates High Confidence Using a Single Microbial Source Tracking Marker.

Microbial source tracking can identify waterbodies at risk of contamination using host-associated molecular markers. No assay used for microbial source tracking is both 100% host-specific and sensitive for human or animal fecal contamination. Using literature sensitivity and specificity values, Bayes' Theorem for conditional probability was applied to the human fecal-associated HF183 marker in a microbial source tracking context. Type I and Type II error rates were examined across a range of priors. Conditional probabilities were investigated using two human-associated markers, HF183 and HumM2, concurrently. Cumulative probability analysis was used to explore the likelihood of true contaminant detection using multiple samples. Probability of human fecal contamination was calculated for all combinations of positive and negative marker results given three samples. Results demonstrate the respective influence that specificity and sensitivity values exert on the likelihood of true positive and true negative. Using practical priors, high levels of confidence (99%) in results were observed when HF183 and HumM2 were used concurrently. Cumulative probability analyses showed that multiple samples from a single location can provide a >95% level of confidence in positive and negative results, suggesting that when multiple samples are necessary to account for in situ variability, a single marker can yield sufficiently reliable results.

One Hundred Fifty Liver Transplants at a New Program in the Modern Era: The University of Mississippi Experience.

From 1991 to 2013, Mississippi was without liver transplant services. In 2013, a new liver transplant program was established at the University of Mississippi Medical Center. Here, we describe our experience with the first 150 transplants over a 4.5-year period. This study is a review of 147 patients who underwent the first 150 liver transplants at the University of Mississippi Medical Center between March 5, 2013, and January 4, 2018. There were no exclusion criteria for this study. Donor, recipient, and outcome variables were analyzed. Recipients were 46% female and 74% white. Age at the time of transplant was 57 [IQR 49-63]. BMI at transplant was 30 [IQR 25-35]. Thirty per cent of transplants were for alcoholic cirrhosis, 25% non-alcoholic steatohepatitis, 24% hepatitis C, and 12% cholestatic. Mean model for end-stage liver disease (MELD) at the time of transplant was 20 [95% confidence interval 19-21] and MELD-Na was 22 [95% confidence interval 20-23]. One-year patient- and graft survival were 89% and 87%, respectively, which were as expected based on Scientific Registry of Transplant Recipient reports after risk adjustment. The data published here verifies it is possible to establish a new liver transplant center in an underserved area previously lacking comprehensive liver care and to achieve results similar to other high-volume centers across the country.

Genome-Wide Transcriptome Landscape of Embryonic Brain-Derived Neural Stem Cells Exposed to Alcohol with Strain-Specific Cross-Examination in BL6 and CD1 Mice.

We have previously reported the deregulatory impact of ethanol on global DNA methylation of brain-derived neural stem cells (NSC). Here, we conducted a genome-wide RNA-seq analysis in differentiating NSC exposed to different modes of ethanol exposure. RNA-seq results showed distinct gene expression patterns and canonical pathways induced by ethanol exposure and withdrawal. Short-term ethanol exposure caused abnormal up-regulation of synaptic pathways, while continuous ethanol treatment profoundly affected brain cells' morphology. Ethanol withdrawal restored the gene expression profile of differentiating NSC without rescuing impaired expression of epigenetics factors. Ingenuity Pathway Analysis (IPA) analysis predicated that ethanol may impact synaptic functions via GABA receptor signalling pathway and affects neural system and brain morphology. We identified Sptbn2, Dcc, and Scn3a as candidate genes which may link alcohol-induced neuronal morphology to brain structural abnormalities, predicted by IPA analysis. Cross-examination of Scn3a and As3mt in differentiated NSC from two different mouse strains (BL6 and CD1) showed a consistent pattern of induction and reduction, respectively. Collectively, our study identifies genetic networks, which may contribute to alcohol-mediated cellular and brain structural dysmorphology, contributing to our knowledge of alcohol-mediated damage to central nervous system, paving the path for better understanding of FASD pathobiology.

Overview of the Genetic Basis and Epigenetic Mechanisms that Contribute to FASD Pathobiology.

Prenatal alcohol (ethanol) exposure (PAE) is the underlying cause for a variety of birth defects and neurodevelopmental deficits referred to as "Fetal Alcohol Spectrum Disorders (FASD)". The more visible phenotypes caused by PAE include growth retardation, and characteristic craniofacial abnormalities associated with functional and structural damage to the central nervous system. Ethanol is a teratogenic agent itself; but it can also alter gene expression. These changes may contribute to the spectrum of effects and different phenotypes that are dependent on alcohol metabolism, as well as the timing and duration of alcohol exposure. Evidence from both human patients and animal models show that genetic factors and epigenetic mechanisms such as DNA methylation, histone post-translational modifications and noncoding RNAs, contribute to the gene expression changes caused by ethanol. Not all embryos that are exposed to alcohol during development exhibit FASD symptoms after birth. FASD patients may present severe birth defects, while others are normal in physical appearance but present a variety of cognitive and behavioral difficulties. It has been hypothesized that maternal and paternal genetic factors may contribute to the sensitivity, resistance or vulnerability of the fetus to alcohol. Moreover, the epigenome is highly sensitive to a multitude of environmental insults including PAE. Studies also show 'transgenerational' effects of alcohol. In such cases, maternal or paternal preconception alcohol consumption could lead to FASD-like phenotypes in the newborn. Thus, the phenotypes in FASD can be modified by interplay between maternal/paternal genetic factors and epigenetic mechanisms. This current review summarizes the contribution of genetic and epigenetic mechanisms in FASD pathobiology, and how this information could be utilized for prevention, early diagnosis and potentially treatment of the affected individuals.

Examining the Colonization and Survival of E. coli from Varying Host Sources in Drainage Basin Sediments and Stormwater.

It is widely understood that stormwater drainage has a significant impact on the health of tidal creek systems via regular inputs of runoff from the surrounding watershed. Due to this hydrologic connection, contamination of the upstream drainage basin will have a direct effect on estuaries and tidal creeks that often act as receiving waters. This study focuses on the importance of drainage basin sediments as they enhance the persistence and transport of the fecal indicator bacteria E. coli within a watershed. Experiments presented use microcosm environments with drainage basin sediments and stormwater to investigate E. coli colonization of stagnant waters and to examine the importance of host sources to bacterial survival. A novel method for establishing microcosms using environmental sediments with in situ bacterial populations and sterile overlying waters is used to examine E. coli colonization of the water column in the absence of flow. Colonization of sterile sediment environments also is examined using two common host sources (human and avian). Each experiment uses sediments of varying grain size and organic content to examine the influence of physical characteristics on bacterial prevalence. Results suggest host source of bacteria may be more important to initial bacterial colonization while physical characteristics of drainage basin sediments better explains extended E. coli persistence. Findings also suggest an indirect control of water column bacterial concentration by sediment type and erodibility.

Development and Detection of Kidd Antibodies.

Kidd antibodies have a reputation for causing hemolytic transfusion reactions and hemolytic disease of the fetus and newborn. We present a case of an untransfused male patient who developed anti-Kidd(a) (Jk(a)) antibodies after receiving an allogenic renal transplant. The formation of this antibody was associated with exposure to the Kidd antigen expressed on the tubular epithelium of the transplanted kidney. The 59-year-old white male patient had received a cadaveric renal transplant at our clinic and returned 5 years later with proteinuria and elevated serum creatinine levels, consistent with nephrotic syndrome. We review the expression of Kidd antigens and the development and detection of Kidd antibodies, and discuss the case reports from the literature of Kidd antibodies associated with kidney-graft rejection that suggest Kidd antigens play a role as a minor histocompatibility antigen.