Shedding Light on Dark Chemical Matter: The Discovery of a SARS-CoV-2 Mpro Main Protease Inhibitor through Intensive Virtual Screening and In Vitro Evaluation.

The development of specific antiviral therapies targeting SARS-CoV-2 remains fundamental because of the continued high incidence of COVID-19 and limited accessibility to antivirals in some countries. In this context, dark chemical matter (DCM), a set of drug-like compounds with outstanding selectivity profiles that have never shown bioactivity despite being extensively assayed, appears to be an excellent starting point for drug development. Accordingly, in this study, we performed a high-throughput screening to identify inhibitors of the SARS-CoV-2 main protease (Mpro) using DCM compounds as ligands. Multiple receptors and two different docking scoring functions were employed to identify the best molecular docking poses. The selected structures were subjected to extensive conventional and Gaussian accelerated molecular dynamics. From the results, four compounds with the best molecular behavior and binding energy were selected for experimental testing, one of which presented inhibitory activity with a Ki value of 48 ± 5 µM. Through virtual screening, we identified a significant starting point for drug development, shedding new light on DCM compounds.

Assessing the Influence of Maternal Age in Bovine Embryos and Oocytes: A Model for Human Reproductive Aging.

In the first weeks after fertilization, embryo mortality in cattle is significantly higher. It is well known that the age of the dam is one of the crucial factors affecting the quality of embryos and oocytes in many mammalian species. In older cattle, there are several evidences that embryo quality decreases, due to a decrease in ovarian reserve, a decrease in mtDNA and ATP, a decrease in progesterone levels, and due to susceptibility to genetic mutations. Herein, we intend to provide an updated summary of recent research on the effects of maternal age on embryos and oocytes of domestic cattle which are a widely used model species for human oocytes and early embryonic development.

Public parks in the city of Rio de Janeiro, southeast Brazil, and the risk of parasitosis transmission by freshwater gastropods.

Urban parks are not only important for the wellbeing of the human population, but are also widely considered to be potentially important sites for the conservation of biodiversity. However, they may offer risk parasitic infections, such as schistosomiasis and fascioliasis, which are both transmitted by freshwater snails. The present study investigated the occurrence of freshwater gastropods in urban parks of the Brazilian city of Rio de Janeiro, and their possible infection by helminths of medical-veterinary importance. Gastropods were collected from six parks (2021 - 2022) and examined for the presence of larval helminths. In all, 12 gastropod species from different families were collected: Ampullariidae, Assimineidae, Burnupidae, Lymnaeidae, Physidae, Planorbidae, Succineidae, and Thiaridae. The parasitological examination revealed cercaria of three types in five snail species, with the Pleurolophocerca cercariae type in Melanoides tuberculata (the most abundant species), Echinostoma cercariae in Physella acuta and Pomacea maculata, and Virgulate cercariae, in Pomacea sp. and Pomacea maculata. None of the Biomphalaria tenagophila and Pseudosuccinea columella (the most frequent species) specimens were parasitized by Schistosoma mansoni or Fasciola hepatica, respectively. Even so, some parks may represent a considerable potential risk for transmission of both Schistosoma mansoni and Fasciola hepatica, given the presence of these gastropod vectors and the frequent contact of visitors with the waterbodies.

Antiviral responses induced by Tdap-IPV vaccination are associated with persistent humoral immunity to Bordetella pertussis.

Many countries continue to experience pertussis epidemics despite widespread vaccination. Waning protection after booster vaccination has highlighted the need for a better understanding of the immunological factors that promote durable protection. Here we apply systems vaccinology to investigate antibody responses in adolescents in the Netherlands (N = 14; NL) and the United Kingdom (N = 12; UK) receiving a tetanus-diphtheria-acellular pertussis-inactivated poliovirus (Tdap-IPV) vaccine. We report that early antiviral and interferon gene expression signatures in blood correlate to persistence of pertussis-specific antibody responses. Single-cell analyses of the innate response identified monocytes and myeloid dendritic cells (MoDC) as principal responders that upregulate antiviral gene expression and type-I interferon cytokine production. With public data, we show that Tdap vaccination stimulates significantly lower antiviral/type-I interferon responses than Tdap-IPV, suggesting that IPV may promote antiviral gene expression. Subsequent in vitro stimulation experiments demonstrate TLR-dependent, IPV-specific activation of the pro-inflammatory p38 MAP kinase pathway in MoDCs. Together, our data provide insights into the molecular host response to pertussis booster vaccination and demonstrate that IPV enhances innate immune activity associated with persistent, pertussis-specific antibody responses.

The Use of Herbal Medicines Among Cancer Patients.

Background and objective The use of herbal medicines has been increasing among cancer patients, as a way to control cancer and treatment-related symptoms; however, many patients are reluctant to disclose this use to their medical practitioners. The fact that oncological treatments have a narrow therapeutic margin, associated with the lack of control and clinical evidence concerning these supplements, makes medication-herbal interactions a reality. These interactions could lead to increased toxicity or a decreased effectiveness of oncological treatment. In light of this, we aimed to assess the prevalence of herbal medicine use in a patient population at a Portuguese central hospital: Centro Hospitalar Lisboa Ocidental. Materials and methods Patients with breast, prostate, or colorectal cancer diagnoses between August 2022 and July 2023 and undergoing oncological treatment were included. Data were collected through a survey during their first appointment, as well as by consulting the patients' clinical files. An interaction evaluation was carried out to assess potential medication-herbal interactions. Finally, a statistical analysis was performed to identify predictive factors for the use of herbal medicines. Results Among the 65 patients included in the study, 52% were females, and the median age of the cohort was 65 years. Breast cancer was the most prevalent diagnosis and the majority of the patients were undergoing palliative treatment. We found that 46% of patients used herbal medicines regularly: to strengthen the immune system, detoxification of the body, and treat insomnia and constipation. A medication-herbal interaction was found in 37% of the cases, the most frequent being doxorubicin-vitamin C, through an antioxidant mechanism. The univariable analysis failed to show any predictive factors associated with the use of herbal medicines. Conclusions This study sheds light on herbal medicine use among cancer patients and the reality of medication-herbal interactions. There is an urgent need for further research and evidence-based medical protocols regarding herbal medicine use, especially in complex cases such as cancer patients, to provide better and safer care.

Are images worth a thousand words? A preliminary study testing a video for owner education in canine atopic dermatitis.

BACKGROUND: Successful management of canine atopic dermatitis (cAD) is challenging and effective pet owner education is crucial to successful outcomes. However, there are limited proven educational strategies in this area. Our goal was to create an effective and engaging educational tool for owners of dogs with cAD. HYPOTHESIS: Video-based education efficacy would be comparable to traditional verbal delivery. Secondary objectives included assessing client perception of the intervention, and determining if there were clinical benefits for the dogs and improved client adherence to treatment. SUBJECTS: Twenty-nine dogs with cAD and their owners were recruited from a teaching hospital of a European veterinary medicine faculty. MATERIALS AND METHODS: In this 8 week, prospective, randomised controlled study, clients in the control group (CG, n = 13) received verbal education and those in the intervention group (IG, n = 16) watched a video. Client knowledge was assessed at Day (D)0 and D56. Treatment adherence and perceived utility and appeal ratings were measured at D56. Clinical progress was assessed at D0 and D56 using CADESI-04 and PVAS10. RESULTS: The differences found in the means of cAD knowledge score, clinical outcomes, utility and appeal ratings and owners' adherence score between groups were not statistically significant. A significant association between the outcome and the intervention group concerning education success [CG, six of 13 (46.15%); IG, 15 of 16 (93.75%)] was found (p = 0.01). CONCLUSIONS AND CLINICAL RELEVANCE: Video-based instructions positively impacted owners' education and demonstrated their potential as a valuable tool. The authors believe that video-based education could be a time-efficient alternative for initial cAD education in veterinary clinics.

Primary Prevention of Canine Atopic Dermatitis: Breaking the Cycle-A Narrative Review.

Canine atopic dermatitis (cAD) is a common and distressing skin condition in dogs, affecting up to 30% of the canine population. It not only impacts their quality of life but also that of their owners. Like human atopic dermatitis (hAD), cAD has a complex pathogenesis, including genetic and environmental factors. Current treatments focus on managing clinical signs, but they can be costly and have limitations. This article emphasizes the importance of preventing cAD from developing in the first place. Understanding the role of the skin's protective barrier is crucial, as its dysfunction plays a vital role in both hAD and cAD. hAD prevention studies have shown promising results in enhancing the skin barrier, but more research is needed to support more robust conclusions. While hAD primary prevention is currently a focal point of intensive investigation in human medicine, research on cAD primary prevention remains under-researched and almost non-existent. Pioneering effective prevention strategies for cAD holds immense potential to enhance the quality of life for both dogs and their owners. Additionally, it bears the promise of a translational impact on human research. Hence, further exploration of this crucial topic is not only relevant but also timely and imperative, warranting support and encouragement.

The Successful Treatment of a Case of Prostate Cancer With Brain Metastasis at Diagnosis.

Brain metastasis in prostate cancer is quite a rare entity, especially when it manifests at diagnosis. The symptoms are usually non-focal and vary based on the location affected. It is almost always associated with a poor prognosis, with an overall survival of less than a year. The ideal management modality for these patients is not well established but a combination of surgery, radiation, and chemotherapy may be possible options based on the extent and systemic involvement. Brain screening is not done systematically in prostate cancer and more research is needed to understand the outcome this decision would lead to. We report a case of a patient diagnosed with prostate cancer with single metastasis to the brain that manifested as headache and vomiting. The patient was treated with surgery, adjuvant irradiation of the surgical bed, and androgen deprivation therapy. He later underwent intensity-modulated radiation therapy (IMRT) to the prostate and has been remarkably relapse-free for four years.

Anxiety and Depression Symptoms During the COVID-19 Pandemic: A Cluster Analysis of Individuals Living in Portugal.

INTRODUCTION: The COVID-19 pandemic is an illustration of how a physical illness can damage people's minds. In this regard, the goal of this study was to see how different sociodemographic and behavioral factors were linked to anxiety and depression symptoms in a group of individuals living in Portugal. METHODS: Between November 2020 and February 2021, a cross-sectional, snowball online study was conducted. The study's target population was adults over the age of 18, residents of the country. For the statistical analysis, the clustering technique - K-means algorithm was applied. The chi-squared test was used to determine the relationships between clusters and sociodemographic and behavioral characteristics. Statistical analyses were conducted in R language, with a significance level of 0.05. A total of 453 participants were included. RESULTS: The majority were female (69.8%), under the age of 40 (60.8%), with a higher education degree (75.3%), and not married (54.4%). Furthermore, the majority were from the country's north region (66%). Cluster 1 (n = 194) was characterized by low or nonexistent levels of anxiety and depression symptoms, which means normal; cluster 2 by severe symptoms (n = 82), meaning case; and cluster 3 by mild symptoms (n = 177), which means borderline. Younger participants (p-value 0.024), female (p-value 0.041), with drinking habits (p-value 0.002), food insecurity (p-value < 0.001), food affordability exacerbation (p-value < 0.001), comorbidity (p-value < 0.001), use of anxiolytics (p-value < 0.001), insufficient household income (p-value 0.017) and income change (p-value < 0.001) were significantly associated with the anxiety-depression clusters. From the three clusters, cluster 2 was mainly represented by younger participants, with more persons stating that their household income was insufficient and that their income has changed as a result of COVID-19 and that they had the highest probability of food insecurity. CONCLUSION: The impacts of a crisis on mental health extend longer than the event itself. We were able to observe that younger women with insufficient household income who suffered a change in income due to COVID-19 and were classified as food insecure presented higher levels of anxiety and depression symptoms. These results highlight the presence of a social gradient where we saw that people who were less advantaged in terms of socioeconomic position presented worse mental health outcomes, stressing, in this sense, the need to bring the best public health responses for these specific groups of the population.

Using a Dual CRISPR/Cas9 Approach to Gain Insight into the Role of LRP1B in Glioblastoma.

LRP1B remains one of the most altered genes in cancer, although its relevance in cancer biology is still unclear. Recent advances in gene editing techniques, particularly CRISPR/Cas9 systems, offer new opportunities to evaluate the function of large genes, such as LRP1B. Using a dual sgRNA CRISPR/Cas9 gene editing approach, this study aimed to assess the impact of disrupting LRP1B in glioblastoma cell biology. Four sgRNAs were designed for the dual targeting of two LRP1B exons (1 and 85). The U87 glioblastoma (GB) cell line was transfected with CRISPR/Cas9 PX459 vectors. To assess LRP1B-gene-induced alterations and expression, PCR, Sanger DNA sequencing, and qRT-PCR were carried out. Three clones (clones B9, E6, and H7) were further evaluated. All clones presented altered cellular morphology, increased cellular and nuclear size, and changes in ploidy. Two clones (E6 and H7) showed a significant decrease in cell growth, both in vitro and in the in vivo CAM assay. Proteomic analysis of the clones' secretome identified differentially expressed proteins that had not been previously associated with LRP1B alterations. This study demonstrates that the dual sgRNA CRISPR/Cas9 strategy can effectively edit LRP1B in GB cells, providing new insights into the impact of LRP1B deletions in GBM biology.

A deep learning approach to estimate x-ray scatter in digital breast tomosynthesis: From phantom models to clinical applications.

BACKGROUND: Digital breast tomosynthesis (DBT) has gained popularity as breast imaging modality due to its pseudo-3D reconstruction and improved accuracy compared to digital mammography. However, DBT faces challenges in image quality and quantitative accuracy due to scatter radiation. Recent advancements in deep learning (DL) have shown promise in using fast convolutional neural networks for scatter correction, achieving comparable results to Monte Carlo (MC) simulations. PURPOSE: To predict the scatter radiation signal in DBT projections within clinically-acceptable times and using only clinically-available data, such as compressed breast thickness and acquisition angle. METHODS: MC simulations to obtain scatter estimates were generated from two types of digital breast phantoms. One set consisted of 600 realistically-shaped homogeneous breast phantoms for initial DL training. The other set was composed of 80 anthropomorphic phantoms, containing realistic internal tissue texture, aimed at fine tuning the DL model for clinical applications. The MC simulations generated scatter and primary maps per projection angle for a wide-angle DBT system. Both datasets were used to train (using 7680 projections from homogeneous phantoms), validate (using 960 and 192 projections from the homogeneous and anthropomorphic phantoms, respectively), and test (using 960 and 48 projections from the homogeneous and anthropomorphic phantoms, respectively) the DL model. The DL output was compared to the corresponding MC ground truth using both quantitative and qualitative metrics, such as mean relative and mean absolute relative differences (MRD and MARD), and to previously-published scatter-to-primary (SPR) ratios for similar breast phantoms. The scatter corrected DBT reconstructions were evaluated by analyzing the obtained linear attenuation values and by visual assessment of corrected projections in a clinical dataset. The time required for training and prediction per projection, as well as the time it takes to produce scatter-corrected projection images, were also tracked. RESULTS: The quantitative comparison between DL scatter predictions and MC simulations showed a median MRD of 0.05% (interquartile range (IQR), -0.04% to 0.13%) and a median MARD of 1.32% (IQR, 0.98% to 1.85%) for homogeneous phantom projections and a median MRD of -0.21% (IQR, -0.35% to -0.07%) and a median MARD of 1.43% (IQR, 1.32% to 1.66%) for the anthropomorphic phantoms. The SPRs for different breast thicknesses and at different projection angles were within ± 15% of the previously-published ranges. The visual assessment showed good prediction capabilities of the DL model with a close match between MC and DL scatter estimates, as well as between DL-based scatter corrected and anti-scatter grid corrected cases. The scatter correction improved the accuracy of the reconstructed linear attenuation of adipose tissue, reducing the error from -16% and -11% to -2.3% and 4.4% for an anthropomorphic digital phantom and clinical case with similar breast thickness, respectively. The DL model training took 40 min and prediction of a single projection took less than 0.01 s. Generating scatter corrected images took 0.03 s per projection for clinical exams and 0.16 s for one entire projection set. CONCLUSIONS: This DL-based method for estimating the scatter signal in DBT projections is fast and accurate, paving the way for future quantitative applications.

Case Report: Patient with deficiency of ADA2 presenting leukocytoclastic vasculitis and pericarditis during infliximab treatment.

Deficiency of adenosine deaminase 2 (DADA2), first reported in 2014, is a disease with great phenotypic variability, which has been increasingly reported. Therapeutic response depends on the phenotype. We present a case of an adolescent with recurrent fever, oral aphthous ulcers, and lymphadenopathy from 8 to 12 years of age and subsequently presented with symptomatic neutropenia. After the diagnosis of DADA2, therapy with infliximab was started, but after the second dose, she developed leukocytoclastic vasculitis and showed symptoms of myopericarditis. Infliximab was switched to etanercept, with no relapses. Despite the safety of tumor necrosis factor alpha inhibitors (TNFi), paradoxical adverse effects have been increasingly reported. The differential diagnosis between disease new-onset manifestations of DADA2 and side effects of TNFi can be challenging and warrants further clarification.

One Health Approach to Tackle Microbial Contamination on Poultries-A Systematic Review.

This study reports the search of available data published regarding microbial occupational exposure assessment in poultries, following the PRISMA methodology. Air collection through filtration was the most frequently used. The most commonly used passive sampling method was material collection such as dust, cages, soils, sediment, and wastewater. Regarding assays applied, the majority of studies comprised culture-based methods, but molecular tools were also frequently used. Screening for antimicrobial susceptibility was performed only for bacteria; cytotoxicity, virological and serological assays were also performed. Most of the selected studies focused on bacteria, although fungi, endotoxins, and ß-glucans were also assessed. The only study concerning fungi and mycotoxins reported the carcinogenic mycotoxin AFB1. This study gives a comprehensive overview of microbial contamination in the poultry industry, emphasizing this setting as a potential reservoir of microbial pathogens threatening human, animal, and environmental health. Additionally, this research helps to provide a sampling and analysis protocol proposal to evaluate the microbiological contamination in these facilities. Few articles were found reporting fungal contamination in poultry farms worldwide. In addition, information concerning fungal resistance profile and mycotoxin contamination remain scarce. Overall, a One Health approach should be incorporated in exposure assessments and the knowledge gaps identified in this paper should be addressed in further research.

Helicobacter pylori infection induces abnormal expression of pro-angiogenic gene ANGPT2 and miR-203a in AGS gastric cell line.

Helicobacter pylori colonizes the stomach and induces an inflammatory response that can develop into gastric pathologies including cancer. The infection can alter the gastric vasculature by the deregulation of angiogenic factors and microRNAs. In this study, we investigate the expression level of pro-angiogenic genes (ANGPT2, ANGPT1, receptor TEK), and microRNAs (miR-135a, miR-200a, miR-203a) predicted to regulate those genes, using H. pylori co-cultures with gastric cancer cell lines. In vitro infections of different gastric cancer cell lines with H. pylori strains were performed, and the expression of ANGPT1, ANGPT2, and TEK genes, and miR-135a, miR-200a, and miR-203a, was quantified after 24 h of infection (h.p.i.). We performed a time course experiment of H. pylori 26695 infections in AGS cells at 6 different time points (3, 6, 12, 28, 24, and 36 h.p.i.). The angiogenic response induced by supernatants of non-infected and infected cells at 24 h.p.i. was evaluated in vivo, using the chicken chorioallantoic membrane (CAM) assay. In response to infection, ANGPT2 mRNA was upregulated at 24 h.p.i, and miR-203a was downregulated in AGS cells co-cultured with different H. pylori strains. The time course of H. pylori 26695 infection in AGS cells showed a gradual decrease of miR-203a expression concomitant with an increase of ANGPT2 mRNA and protein expression. Expression of ANGPT1 and TEK mRNA or protein could not be detected in any of the infected or non-infected cells. CAM assays showed that the supernatants of AGS-infected cells with 26695 strain induced a significantly higher angiogenic and inflammatory response. Our results suggest that H. pylori could contribute to the process of carcinogenesis by downregulating miR-203a, which further promotes angiogenesis in gastric mucosa by increasing ANGPT2 expression. Further investigation is needed to elucidate the underlying molecular mechanisms.

Assessing the Antitumor Potential of Variants of the Extracellular Carbohydrate Polymer from Synechocystis ΔsigF Mutant.

Cancer is a leading cause of death worldwide with a huge societal and economic impact. Clinically effective and less expensive anticancer agents derived from natural sources can help to overcome limitations and negative side effects of chemotherapy and radiotherapy. Previously, we showed that the extracellular carbohydrate polymer of a Synechocystis ΔsigF overproducing mutant displayed a strong antitumor activity towards several human tumor cell lines, by inducing high levels of apoptosis through p53 and caspase-3 activation. Here, the ΔsigF polymer was manipulated to obtain variants that were tested in a human melanoma (Mewo) cell line. Our results demonstrated that high molecular mass fractions were important for the polymer bioactivity, and that the reduction of the peptide content generated a variant with enhanced in vitro antitumor activity. This variant, and the original ΔsigF polymer, were further tested in vivo using the chick chorioallantoic membrane (CAM) assay. Both polymers significantly decreased xenografted CAM tumor growth and affected tumor morphology, by promoting less compact tumors, validating their antitumor potential in vivo. This work contributes with strategies for the design and testing tailored cyanobacterial extracellular polymers and further strengths the relevance of evaluating this type of polymers for biotechnological/biomedical applications.

Her2 Status Discrepancy Between Core Needle Biopsy and Surgically Resected Mastectomy Specimen: A Clinical Case.

The biomarker concordance between core needle biopsy (CNB) and surgical specimen (SS), in breast cancer, has long been a matter of discussion because of its influence on oncologic treatment choice. Particularly, human epidermal growth factor receptor 2 (Her2) status is quite important, because of the impact on breast cancer classification and target therapy. Many factors could influence the difference in biomarker status between samples, such as the technic itself, sample procedures and intratumoral heterogeneity. Neoadjuvant chemotherapy (NAC) can also contribute to this variation and should be taken into consideration. We report a clinical case of a 33-year-old man who was diagnosed with right breast cancer, initially Her2 negative in the CNB. Therefore, the NAC was completed without anti-Her2 treatment. Later, after the mastectomy, revision of the SS was found to be positive for Her2, after the silver in situ hybridization (SISH) technique. Consequently, the patient lost the chance of doing anti-Her2 therapy in the neoadjuvant setting, reducing his possibility to achieve a complete pathologic response and later jeopardizing his clinical outcome.