A method for isolating and culturing placental cells from failed early equine pregnancies

Early pregnancy loss occurs in 6–10% of equine pregnancies making it the main cause of reproductive wastage. Despite this, reasons for the losses are known in only 16% of cases. Lack of viable conceptus material has inhibited investigations of many potential genetic and pathological causes. We present a method for isolating and culturing placental cells from failed early equine pregnancies. Trophoblast cells from 18/30 (60%) failed equine pregnancies of gestational ages 14–65 days were successfully cultured in three different media, with the greatest growth achieved for cells cultured in AmnioChrome™ Plus. Genomic DNA of a suitable quality for molecular assays was also isolated from 29/30 of these cases. This method will enable future investigations determining pathologies causing EPL

Evaluating the effects of climate change and chemical, physical, and biological stressors on nearshore coral reefs: A case study in the Great Barrier Reef, Australia

An understanding of the combined effects of climate change (CC) and other anthropogenic stressors, such as chemical exposures, is essential for improving ecological risk assessments of vulnerable ecosystems. In the Great Barrier Reef, coral reefs are under increasingly severe duress from increasing ocean temperatures, acidification, and cyclone intensities associated with CC. In addition to these stressors, inshore reef systems, such as the Mackay–Whitsunday coastal zone, are being impacted by other anthropogenic stressors, including chemical, nutrient, and sediment exposures related to more intense rainfall events that increase the catchment runoff of contaminated waters. To illustrate an approach for incorporating CC into ecological risk assessment frameworks, we developed an adverse outcome pathway network to conceptually delineate the effects of climate variables and photosystem II herbicide (diuron) exposures on scleractinian corals. This informed the development of a Bayesian network (BN) to quantitatively compare the effects of historical (1975–2005) and future projected climate on inshore hard coral bleaching, mortality, and cover. This BN demonstrated how risk may be predicted for multiple physical and biological stressors, including temperature, ocean acidification, cyclones, sediments, macroalgae competition, and crown of thorns starfish predation, as well as chemical stressors such as nitrogen and herbicides. Climate scenarios included an ensemble of 16 downscaled models encompassing current and future conditions based on multiple emission scenarios for two 30-year periods. It was found that both climate-related and catchment-related stressors pose a risk to these inshore reef systems, with projected increases in coral bleaching and coral mortality under all future climate scenarios. This modeling exercise can support the identification of risk drivers for the prioritization of management interventions to build future resilient reefs.publishedVersio

Anti-HBV treatment induces novel reverse transcriptase mutations with reflective effect on HBV S antigen

The identification of novel reverse-transcriptase (RT) drug-resistance mutations is critical in predicting the probability of success to anti-HBV treatment. Furthermore, due to HBV-RT/HBsAg gene-overlap, they can have an impact on HBsAg-detection and quantification

Design of a Low‐Power Radio Frequency Unit and Its Application for Bacterial Inactivation under Laboratory Conditions

A lab‐scale low‐power free‐running radio frequency (RF) oscillator operating at a frequency of 27.12 ± 0.50 MHz was developed to be suitable for fundamental microbiological research topics. Calibration and validation were conducted for two common foodborne pathogens in relevant microbiological growth media, i.e., Salmonella Typhimurium and Listeria monocytogenes in Tryptic Soy Broth and Brain–Heart Infusion broth, respectively. The evolution of temperature, frequency, and power consumption was monitored during treatments, both with and without bacterial cells. The setup operated within the predefined frequency range, reaching temperatures of 71–76 °C after 15 min. The average power consumption ranged between 12 and 14 W. The presence of bacteria did not significantly influence the operational parameters. The inactivation potential of the RF setup was validated, demonstrating the absence of viable cells after 8 and 10 min of treatment, for S. Typhimurium and L. monocytogenes, respectively. In future studies, the setup can be used to conduct fundamental microbiological studies on RF inactivation. The setup can provide added value to the scientific field, since (i) no consensus has been reached on the inactivation mechanisms of RF inactivation of pathogens in foods and (ii) most commercial RF setups are unsuitable to adopt for fundamental studies. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Funding: This work was funded by the KU Leuven Research Fund through project C24/18/046, by the Research Foundation Flanders (FWO) through project G0B4121N, and by the EU H2020 research and innovation program under the Marie Skłodowska‐Curie grant agreement no. 956126. Authors Davy Verheyen and Simen Akkermans were funded by the Research Foundation Flanders (FWO), grant numbers 1254421N and 1224620N, respectively

Novel HBsAg markers tightly correlate with occult HBV infection and strongly affect HBsAg detection.

Occult HBV infection (OBI) is a threat for the safety of blood-supply, and has been associated with the onset of HBV-related hepatocellular carcinoma and lymphomagenesis. Nevertheless, genetic markers in HBsAg (particularly in D-genotype, the most common in Europe) significantly associated with OBI in vivo are missing. Thus, the goal of this study is to define: (i) prevalence and clinical profile of OBI among blood-donors; (ii) HBsAg-mutations associated with OBI; (iii) their impact on HBsAg-detection. OBI was searched among 422,278 blood-donors screened by Nucleic-Acid-Testing. Following Taormina-OBI-definition, 26 (0.006%) OBI-patients were identified. Despite viremia <50IU/ml, HBsAg-sequences were obtained for 25/26 patients (24/25 genotype-D). OBI-associated mutations were identified by comparing OBI-HBsAg with that of 82 chronically-infected (genotype-D) patients as control. Twenty HBsAg-mutations significantly correlated for the first time with OBI. By structural analysis, they localized in the major HBV B-cell-epitope, and in HBsAg-capsid interaction region. 14/24 OBI-patients (58.8%) carried in median 3 such mutations (IQR:2.0-6.0) against 0 in chronically-infected patients. By co-variation analysis, correlations were observed for R122P+S167L (phi=0.68, P=0.01), T116N+S143L (phi=0.53, P=0.03), and Y100S+S143L (phi=0.67, p<0.001). Mutants (obtained by site-directed mutagenesis) carrying T116N, T116N+S143L, R122P, R122P+Q101R, or R122P+S167L strongly decreased HBsAg-reactivity (54.9±22.6S/CO, 31.2±12.0S/CO, 6.1±2.4S/CO, 3.0±1.0S/CO and 3.9±1.3S/CO, respectively) compared to wild-type (306.8±64.1S/CO). Even more, Y100S and Y100S+S143L supernatants show no detectable-HBsAg (experiments in quadruplicate). In conclusions, unique HBsAg-mutations in genotype-D, different than those described in genotypes B/C (rarely found in western countries), tightly correlate with OBI, and strongly affect HBsAg-detection. By altering HBV-antigenicity and/or viral-particle maturation, they may affect full-reliability of universal diagnostic-assays for HBsAg-detection

Descriptive study of current therapeutic practices, clinical reproductive findings and incidence of pregnancy loss in intensively managed thoroughbred mares

Therapeutic practices in equine reproductive medicine have dramatically evolved over the last 20 years but current usage is not described. The aims of this study were to provide a description of medication use and clinical findings of reproductive examinations alongside measures of reproductive efficiency in thoroughbreds. A prospective cohort study was conducted in the 2013 and 2014 breeding seasons. Mare and stallion details, information on veterinary interventions and findings of reproductive ultrasound scans were collected using questionnaires and entered into a custom-designed Microsoft Access database. Descriptive summary statistics were derived directly from the database and using Microsoft Excel. Information was collected from 2246 pregnancies in 1754 mares from 29 stud farms. Ovulatory induction agents were used in 91.8% of cases, oestrus induction agents in 38.4% and covering therapies in 62.7%. Intrauterine anti-microbials were used in 49.6% of mares. Single pregnancies accounted for 83.9% of pregnancies, twins for 15.3% and triplets for 0.7%. The overall incidence of pregnancy loss between days 15-42 was 6.4% (95% CI 5.4%, 7.4%) and 1.6% (95% CI 1.1%, 2.1%) between days 43-65. A further 1.3% of pregnancies were lost by October and 4.5% by birth (including stillbirths). Eighty-three percent of all pregnancies resulted in a live foal. In conclusion, there has been a considerable increase in the use of reproductive therapeutics over the last 12 years. Nonetheless, incidence of pregnancy loss and live foal percentages remain essentially unchanged. Risk factor studies are required to determine if the substantial increase in therapeutic usage is conferring positive benefits

Improved Bevirimat resistance prediction by combination of structural and sequence-based classifiers

<p>Abstract</p> <p>Background</p> <p>Maturation inhibitors such as Bevirimat are a new class of antiretroviral drugs that hamper the cleavage of HIV-1 proteins into their functional active forms. They bind to these preproteins and inhibit their cleavage by the HIV-1 protease, resulting in non-functional virus particles. Nevertheless, there exist mutations in this region leading to resistance against Bevirimat. Highly specific and accurate tools to predict resistance to maturation inhibitors can help to identify patients, who might benefit from the usage of these new drugs.</p> <p>Results</p> <p>We tested several methods to improve Bevirimat resistance prediction in HIV-1. It turned out that combining structural and sequence-based information in classifier ensembles led to accurate and reliable predictions. Moreover, we were able to identify the most crucial regions for Bevirimat resistance computationally, which are in line with experimental results from other studies.</p> <p>Conclusions</p> <p>Our analysis demonstrated the use of machine learning techniques to predict HIV-1 resistance against maturation inhibitors such as Bevirimat. New maturation inhibitors are already under development and might enlarge the arsenal of antiretroviral drugs in the future. Thus, accurate prediction tools are very useful to enable a personalized therapy.</p