Cytoskeletal ß-tubulin and cysteine cathepsin L deregulation by SARS-CoV-2 spike protein interaction with the neuronal model cell line SH-SY5Y.

SARS-CoV-2 mainly infects the respiratory tract but can also target other organs, including the central nervous system. While it was recently shown that cells of the blood-brain-barrier are permissive to SARS-CoV-2 infection in vitro, it remains debated whether neurons can be infected. In this study, we demonstrate that vesicular stomatitis virus particles pseudotyped with the spike protein of SARS-CoV-2 variants WT, Alpha, Delta and Omicron enter the neuronal model cell line SH-SY5Y. Cell biological analyses of the pseudo-virus treated cultures showed marked alterations in microtubules of SH-SY5Y cells. Because the changes in ß-tubulin occurred in most cells, but only few were infected, we further asked whether interaction of the cells with spike protein might be sufficient to cause molecular and structural changes. For this, SH-SY5Y cells were incubated with trimeric spike proteins for time intervals of up to 24 h. CellProfiler™-based image analyses revealed changes in the intensities of microtubule staining in spike protein-incubated cells. Furthermore, expression of the spike protein-processing protease cathepsin L was found to be up-regulated by wild type, Alpha and Delta spike protein pseudotypes and cathepsin L was found to be secreted from spike protein-treated cells. We conclude that the mere interaction of the SARS-CoV-2 with neuronal cells can affect cellular architecture and proteolytic capacities. The molecular mechanisms underlying SARS-CoV-2 spike protein induced cytoskeletal changes in neuronal cells remain elusive and require future studies.

Myositis Ossificans of the Breast Mimicking Breast Cancer: A Case Report.

Myositis ossificans (MO) is an uncommon tumor characterized by a rapidly growing mass following a history of local trauma. Few cases of MO affecting the breast have been reported, and some were misdiagnosed as primary osteosarcoma of the breast or metaplastic breast carcinoma. The following case report presents a patient with a growing breast lump whose core biopsy result was suspicious for breast cancer. MO was diagnosed after analysis of the mastectomy specimen. This case highlights the importance of MO as a differential diagnosis of a growing soft-tissue mass after trauma to avoid unnecessary overtreatment. Keywords: Myositis Ossificans, Osteosarcoma, Breast Cancer, Mastectomy, Heterotopic Ossification © RSNA, 2023.

Metabolomics characterizes early metabolic changes and markers of tolerant Eucalyptus ssp. clones against drought stress.

EUCALYPTUS: L'Hér. (Myrtaceae) is one of the economically most important and widely cultivated trees for wood crop purposes worldwide. Climatic changes together with the constant need to expand plantations to areas that do not always provide optimal conditions for plant growth highlight the need to assess the impact of abiotic stresses on eucalypt trees. We aimed to unveil the drought effect on the leaf metabolome of commercial clones with differential phenotypic response to this stress. For this, seedlings of 13 clones were grown at well-watered (WW) and water-deficit (WD) conditions and their leaf extracts were subjected to comparative analysis using ultra-high performance liquid chromatography coupled to mass spectrometry (UPLC-MS) and nuclear magnetic resonance spectroscopy (NMR). UPLC-MS and NMR analyses led to the annotation of over 100 molecular features of classes such as cyclitols, phenolics, flavonoids, formylated phloroglucinol compounds (FPCs) and fatty acids. Multivariate data analysis was employed for specimens' classifications and markers identification from both platforms. The results obtained in this work allowed us to classify clones differing in drought tolerance. Classification models were validated using an extra subset of samples. Tolerant plants exposed to water deficit accumulated arginine, gallic acid derivatives, caffeic acid and tannins at higher levels. In contrast, stressed drought-sensitive clones were characterised by a significant reduction in glucose, inositol and shikimic acid levels. These changes in contrasting drought response eucalypt pave ways for differential outcomes of tolerant and susceptible phenotypes. Under optimal growth conditions, all clones were rich in FPCs. These results can be used for early screening of tolerant clones and to improve our understanding of the role of these biomarkers in Eucalyptus tolerance to drought stress.

Quantifying and mapping species threat abatement opportunities to support national target setting.

The successful implementation of the Convention on Biological Diversity's post-2020 Global Biodiversity Framework will rely on effective translation of targets from global to national level and increased engagement across diverse sectors of society. Species conservation targets require policy support measures that can be applied to a diversity of taxonomic groups, that link action targets to outcome goals, and that can be applied to both global and national data sets to account for national context, which the species threat abatement and restoration (STAR) metric does. To test the flexibility of STAR, we applied the metric to vascular plants listed on national red lists of Brazil, Norway, and South Africa. The STAR metric uses data on species' extinction risk, distributions, and threats, which we obtained from national red lists to quantify the contribution that threat abatement and habitat restoration activities could make to reducing species' extinction risk. Across all 3 countries, the greatest opportunity for reducing plant species' extinction risk was from abating threats from agricultural activities, which could reduce species' extinction risk by 54% in Norway, 36% in South Africa, and 29% in Brazil. Species extinction risk could be reduced by a further 21% in South Africa by abating threats from invasive species and by 21% in Brazil by abating threats from urban expansion. Even with different approaches to red-listing among countries, the STAR metric yielded informative results that identified where the greatest conservation gains could be made for species through threat-abatement and restoration activities. Quantifiably linking local taxonomic coverage and data collection to global processes with STAR would allow national target setting to align with global targets and enable state and nonstate actors to measure and report on their potential contributions to species conservation.

Zinc(II) complexes bearing N,N,S ligands: Synthesis, crystal structure, spectroscopic analysis, molecular docking and biological investigations about its antifungal activity.

In the present work, the synthesis, characterization, antifungal activity, molecular docking study and in silico approach of five thiosemicarbazone derivatives and their corresponding zinc(II) complexes are described. The compounds were characterized by elemental analysis, IR, UV-Vis and NMR spectroscopic measurements, molar conductivity measurements, emission spectra, high-resolution mass spectrometry and X ray study. The antifungal activity of the free ligands and synthesized compounds was preliminarily evaluated against Candida albicans (ATCC 90028), Candida tropicalis (ATCC 13803) and Candida glabrata (ATCC 2001), by the minimum inhibitory concentration (MIC) assay. Two complexes, 4 (MIC = 3.18 to 6.37 µM) and 5 (MIC = 25.95 µM for all) showed promising results, being highly active against all strains evaluated. The X-ray analyses shown that the complex 2 crystallizes in the centrosymmetric space group P21/c of the monoclinic system and the coordination sphere around zinc(II) atom is better described as slightly distorted octahedral. The Hirshfeld surface (HS) analysis showed that non-classical H···H and C···H/H···C contacts contribute with 65.9% while the S···H and N···H (21%) and Cl···H and O···H interactions (12%) complete the HS area. The molecular docking results, performed against CYP51 enzyme (sterol 14α-demethylase) of C. albicans and C. glabrata shows that the complexes 4 (ΔG = -10.75 and - 12.90 kcal/ mol) and 5 (ΔG = -11.12 and - 14.53 kcal/ mol) showed the highest binding free energies of all compounds. The ADME-Tox (absorption, distribution, metabolism, excretion and toxicity) in silico parameters evaluated showed promising results for all compounds.

Predictors of parental satisfaction with the pediatric ophthalmology care in a Brazilian hospital.

The purpose of this cross-sectional study was to assess the satisfaction of parents of visually impaired children with the pediatric ophthalmology care in a Brazilian hospital and to associate parental satisfaction with predictor variables. The study enrolled children with a binocular and monocular visual impairment aged between 4 and 17 years from a single site in Brazil between June 2019 and May 2020. Parents or legal guardians answered the Brazilian version of the Pediatric Quality of Life InventoryTM (PedsQLTM 3.0) Healthcare Satisfaction Generic - Parents. Linear regression analysis was used to investigate the association between the PedsQL scores and predictor variables. The sample included 82 children and their respective parents. The mean PedsQL score was 90.3 ± 6.6 (out of 100). In the univariate model, child female sex (p = 0.056), child older age (p = 0.078), better eye visual acuity (VA; p = 0.016), and worse eye VA (p = 0.008) were significantly associated with higher scores. The multivariate estimates identified the worse eye VA (p = 0.034) as a predictor of parental satisfaction. In summary, parents' satisfaction with this pediatric ophthalmology care was high. Nevertheless, a continuous evaluation is warranted to keep up with parents' expectations and plan for possible improvements in clinical practice.

Benefits of pharmacological and electrical cholinergic stimulation in hypertension and heart failure.

Systemic arterial hypertension and heart failure are cardiovascular diseases that affect millions of individuals worldwide. They are characterized by a change in the autonomic nervous system balance, highlighted by an increase in sympathetic activity associated with a decrease in parasympathetic activity. Most therapeutic approaches seek to treat these diseases by medications that attenuate sympathetic activity. However, there is a growing number of studies demonstrating that the improvement of parasympathetic function, by means of pharmacological or electrical stimulation, can be an effective tool for the treatment of these cardiovascular diseases. Therefore, this review aims to describe the advances reported by experimental and clinical studies that addressed the potential of cholinergic stimulation to prevent autonomic and cardiovascular imbalance in hypertension and heart failure. Overall, the published data reviewed demonstrate that the use of central or peripheral acetylcholinesterase inhibitors is efficient to improve the autonomic imbalance and hemodynamic changes observed in heart failure and hypertension. Of note, the baroreflex and the vagus nerve activation have been shown to be safe and effective approaches to be used as an alternative treatment for these cardiovascular diseases. In conclusion, pharmacological and electrical stimulation of the parasympathetic nervous system has the potential to be used as a therapeutic tool for the treatment of hypertension and heart failure, deserving to be more explored in the clinical setting.

Wireworm (Coleoptera: Elateridae) Species Composition and Management in Sweet Potato Grown in North Florida Using Chemical Insecticides and Entomopathogenic Nematodes.

Wireworms are immature stages of click beetles (Coleoptera: Elateridae) and are considered a serious threat to sweet potato production in the southern United States. The major wireworm species collected in North Florida sweet potato fields in 2017 and 2018 were Conoderus scissus, C. rudis, C. amplicollis, and C. falli. These species vary in their behavior and biology. During a 2-yr study period, we conducted two insecticide field trials using eleven insecticides belonging to organophosphates, neonicotinoids, pyrethroids, and botanical classes, and three field trials using entomopathogenic nematode (EPN) species to control wireworms. In 2017, all insecticide treatments significantly reduced new feeding holes and total holes (old + new + other) as compared to the untreated control. In 2018, the result was similar with a few variations. In both years, all insecticides showed a percentage reduction in wireworm damage holes (2017: 34.88-96.19%; 2018: 12.38-97.02%) with the highest by Regent. In the EPN field study, one application of EPN near planting significantly reduced soil insects. In a laboratory study conducted at the Tropical Research and Education Center, UF-IFAS, chlorpyrifos caused higher percentage mortality of C. rudis (55.5%) than C. scissus (22.2%). At the present experiment rates, none of the insecticides caused the mortality of C. amplicollis. Heterorhabditids strain 'FL-2122' was more susceptible to chlorpyrifos than other strains of EPN.

Prognostic and predictive value of Pleckstrin homology-like domain, family A family members in breast cancer.

Aim: The PHLDA (pleckstrin homology like domain, family A) gene family encodes proteins capable of inhibiting AKT (serine/threonine kinase) signaling through phosphoinositol binding competition. Results & methodology: Using in silico analysis, we found that Luminal A and B patients' short relapse-free survival was associated with low PHLDA1 or PHLDA3 and high PHLDA2 expression. In a cohort of 393 patients with luminal breast cancer evaluated by immunohistochemistry on tissue microarrays, we found a direct association of PHLDA3 expression with hormonal therapy response (p = 0.013). Conclusion: Our findings provide new information on the role played by the PHLDA family members as prognostic markers in breast cancer, and more importantly, we provide evidence that they might also predict a response to endocrine therapy.

Variability - Surface intensity distribution of large-area reference sources.

In this paper we present the impact of variability, a surface source parameter, on the efficiency evaluation of surface contamination monitors. This study was based on two source uniformity correction methodologies and data from real surface source distributions. Surface source intensity distribution has been changed by rearranging the cells (portions of the active area of each LARS) while keeping the same source uniformity value. Instrument efficiencies have been calculated for different sets of uniformities and variabilities. This study led to emphasize the importance of variability, a differential source intensity distribution parameter, over the uniformity, an integral source intensity distribution parameter, and reinforced the importance of the source uniformity correction procedure on the course of surface contamination monitor calibration.

Correction factors for non-uniform large-area reference sources.

Based on uniformity measurements of large-area reference sources used in calibration procedures of surface contamination monitors, an investigation was carried out to obtain a method that estimates the bias originated from surface source intensity distribution deviation from the ideal uniform distribution and corrects it. It relies on correcting the estimated instrument efficiency by applying correction factors driven from the uniformity distribution profiles of the sources used in calibration procedure. Simulations of the monitor calibration procedure are run for 2 distinct surface source distributions: the real and the ideally uniform distributions. Correction factors are driven from counting rate estimates obtained from each source representation. In order to evaluate adequacy of this proposition it was validated against a method proposed by the NPL in the Good Practices Guide No.14.

Hepatocyte gene expression and DNA methylation as ancestry-dependent mechanisms in African Americans.

African Americans (AAs) are an admixed population with widely varying proportion of West African ancestry (WAA). Here we report the correlation of WAA to gene expression and DNA methylation in AA-derived hepatocytes, a cell type important in disease and drug response. We perform mediation analysis to test whether methylation is a mediator of the effect of ancestry on expression. GTEx samples and a second cohort are used as validation. One hundred and thirty-one genes are associated with WAA (FDR < 0.10), 28 of which replicate and represent 220 GWAS phenotypes. Among PharmGKB pharmacogenes, VDR, PTGIS, ALDH1A1, CYP2C19, and P2RY1 nominally associate with WAA (p < 0.05). We find 1037 WAA-associated, differentially methylated regions (FDR < 0.05), with hypomethylated genes enriched in drug-response pathways. In conclusion, WAA contributes to variability in hepatocyte expression and DNA methylation with identified genes previously implicated for diseases disproportionately affecting AAs, including cardiovascular (PTGIS, PLAT) and renal (APOL1) disease, and drug response (CYP2C19).

Influence of microhabitat on the richness of anuran species: a case study of different landscapes in the Atlantic Forest of southern Brazil.

Environmental heterogeneity is a factor which can help explain the higher local species richness. The objective of this study was to test if richness and composition of anurans species are related to available microhabitats and landscape type of sampled sites. We assume that a higher number of microhabitats increase environmental heterogeneity and this, in turn, affects species richness of amphibians. We performed the study in the Mesophytic Semideciduous Forest, a vegetation type within Atlantic Forest Domain. Between October 2010 and February 2011, we sampled 23 water bodies located in the agricultural, forest, and urban landscapes. The species richness was determined using survey at breeding sites methodology, and the availability of microhabitats was estimated visually. Thirty-four anuran species belonging to 12 families were recorded. The species richness in water bodies ranged from two to 13 species. The highest species richness was recorded in environments with a higher number of microhabitats, while the species composition in water bodies was partially grouped according to the predominant landscape type that is agricultural, forest, forest edge or urban. Our results suggest that species use specific environments (e.g. landscapes, habitat and microhabitat) for their reproductive activities.

Can acetylcysteine ameliorate cisplatin-induced toxicities and oxidative stress without decreasing antitumor efficacy? A randomized, double-blind, placebo-controlled trial involving patients with head and neck cancer.

The protective antioxidant activity of acetylcysteine (NAC) against toxicity due to cisplatin has been reported in experimental models; however, its efficacy in patients has not been elucidated. The aim of this study was to investigate the possible protective effect of NAC on cisplatin-induced toxicity and the effect of NAC on clinical response and oxidative stress in patients treated for head and neck cancer. This was a randomized, double-blind, placebo-controlled trial conducted in patients receiving high-dose cisplatin chemotherapy concomitant to radiotherapy. Patients were randomly assigned to groups and received: (a) 600 mg NAC syrup, orally once daily at night for 7 consecutive days or (b) placebo, administered similarly to NAC. Nephro-, oto-, hepato-, myelo-, and gastrointestinal toxicities, clinical responses, and plasma and cellular markers of oxidative stress were evaluated. Fifty-seven patients were included (n = 28, NAC arm; and n = 29, placebo arm). A high prevalence of most types of toxicities was observed after cisplatin chemotherapy; however, the parameters were similar between the two groups. There was a predominance of partial response to treatment. In the cellular and plasmatic oxidative stress analyses, minor differences were observed. Overall, there was no statistically significant difference between the groups for all outcomes. These findings show that low-dose oral NAC does not protect patients with head and neck cancer from cisplatin-induced toxicities and oxidative stress. The antitumor efficacy of cisplatin was apparently not impaired by NAC.

Characterization of Dendritic Cell-Derived Extracellular Vesicles During Dengue Virus Infection.

The dengue virus (DENV), transmitted by Aedes spp. mosquitoes, is one of the most important arboviral infections in the world. Dengue begins as a febrile condition, and in certain patients, it can evolve severe clinical outcomes, such as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). The reasons why certain patients develop DHF or DSS have not been thoroughly elucidated to date, and both patient and viral factors have been implicated. Previous work has shown that a severe immune dysfunction involving dendritic cells and T cells plays a key role in increasing the disease severity, especially in secondary heterologous infections. Extracellular vesicles (EVs) are membranous particles that are secreted by several cell types involved in homeostatic and pathological processes. Secretion of EVs by infected cells can enhance immune responses or favor viral evasion. In this study, we compare the molecular content of EVs that are secreted by human primary dendritic cells under different conditions: uninfected or infected with DENV3 strains isolated from patients with different infection phenotypes (a severe case involving DSS and a mild case). Human monocyte-derived dendritic cells (mdDCs) were infected with the dengue virus strains DENV3 5532 (severe) or DENV3 290 (mild), and the EVs were isolated. The presence of cup-shaped EVs was confirmed by electron microscopy and immunostaining with CD9, CD81, and CD83. The RNA content from the mdDC-infected cells contained several mRNAs and miRNAs related to immune responses compared to the EVs from mock-infected mdDCs. A number of these RNAs were detected exclusively during infection with DENV3 290 or DENV3 5532. This result suggests that the differential immune modulation of mdDCs by dengue strains can be achieved through the EV pathway. Additionally, we observed an association of EVs with DENV-infectious particles that seem to be protected from antibodies targeting the DENV envelope protein. We also showed that EVs derived from cells treated with IFN alpha have a protective effect against DENV infection in other cells. These results suggested that during DENV infection, the EV pathway could be exploited to favor viral viability, although immune mechanisms to counteract viral infection can also involve DC-derived EVs.

Fenofibrate reverses changes induced by high-fat diet on metabolism in mice muscle and visceral adipocytes.

The effect of fenofibrate on the metabolism of skeletal muscle and visceral white adipose tissue of diet-induced obese (DIO) mice was investigated. C57BL/6J male mice were fed either a control or high-fat diet for 8 weeks. Fenofibrate (50 mg/Kg BW, daily) was administered by oral gavage during the last two weeks of the experimental period. Insulin-stimulated glucose metabolism in soleus muscles, glucose tolerance test, insulin tolerance test, indirect calorimetry, lipolysis of visceral white adipose tissue, expression of miR-103-3p in adipose tissue, and miR-1a, miR-133a/b, miR-206, let7b-5p, miR-23b-3p, miR-29-3p, miR-143-3p in soleus muscle, genes related to glucose and fatty acid metabolism in adipose tissue and soleus muscle, and proteins (phospho-AMPKα2, Pgc1α, Cpt1b), intramuscular lipid staining, and activities of fatty acid oxidation enzymes in skeletal muscle were investigated. In DIO mice, fenofibrate prevented weight gain induced by HFD feeding by increasing energy expenditure; improved whole body glucose homeostasis, and in skeletal muscle, increased insulin dependent glucose uptake, miR-1a levels, reduced intramuscular lipid accumulation, and phospho-AMPKα2 levels. In visceral adipose tissue of obese mice, fenofibrate decreased basal lipolysis rate and visceral adipocytes hypertrophy, and induced the expression of Glut-4, Irs1, and Cav-1 mRNA and miR-103-3p suggesting a higher insulin sensitivity of the adipocytes. The evidence is presented herein that beneficial effects of fenofibrate on body weight, glucose homeostasis, and muscle metabolism might be related to its action in adipose tissue. Moreover, fenofibrate regulates miR-1a-3p in soleus and miR-103-3p in adipose tissue, suggesting these microRNAs might contribute to fenofibrate beneficial effects on metabolism.

A simple questionnaire to detect chronic kidney disease patients from Long An province screening data in Vietnam.

BACKGROUND: The prevalence of chronic kidney disease (CKD) in rural Vietnam is unknown. We wished to determine the prevalence of CKD and determine whether a simple questionnaire was able to detect individuals at high risk of CKD before expensive confirmatory laboratory testing. METHODS: A cross sectional study was performed. We recruited 2037 participants from 13 communes of Long An province, Vietnam, for CKD screening with urine albumin/creatinine ratio (ACR) measured by immunoturbidimetric method and serum creatinine to estimate glomerular filtration rate (eGFR). CKD was defined as either ACR ≥ 30 mg/g or eGFR MDRD < 60 ml/min/1.73 m2. A two page questionnaire with 23 variables was administered to each participant with queries postulated to be correlated with risk of CKD. RESULTS: Of the 2037 participants, 260 (12.76%) were found to have CKD. Five questionnaire variables (age more than 50, measured hypertension, history of diabetes, history of hypertension, and history of a low salt diet) were correlated with CKD, and used to construct a risk score for CKD. CONCLUSIONS: CKD is common in Vietnam. Our questionnaire and risk score tool can be used to detect individuals with a higher likelihood of CKD, and who can then be more economically screened with routine laboratory confirmatory tests.