Miracle-dz/UniFAR-FTUs huggingface dataset:
Legal Frameworks and Controls for the Protection of Research Animals: A Focus on the Animal Welfare Body with a French Case Study

Simple Summary Worldwide, the conditions governing the use of laboratory animals are based on culture and traditions. Applications of the principle of the three Rs—replacement, reduction, and refinement—aim to foster humane and responsible science and minimize animal harm. In this paper, the European system regulating laboratory animal welfare is examined since it is one of the strictest such systems in the world. Growing public concern over the use of animals for research animal use and the importance of high research animal welfare to the outcomes of studies have pushed institutions to strengthen their protocols for research activities involving animals. The animal welfare body (AWB) is a type of regulatory structure responsible for protecting animal well-being in each stage of experimental research programs. It is a local structure that fulfills oversight and advisory functions on animal housing conditions and care and research procedures involving animals while providing support for the implementation of legal requirements. Since this structure is an innovation and examples of its functioning are scarce, a French case study is described here. Information on AWB members’ functions, tasks and goals is discussed by examining the body’s activity in a French research institute. The purpose of this work is to add knowledge in this field and share information on the scientific community’s commitment to welfare progress for laboratory animals. Abstract In recent years, efforts have been devoted to improving the welfare of laboratory animals. Scientific progress and growing concerns over animal harm have pushed institutions to strengthen their laws to make science more humane and responsible. European Directive 2010/63/EU makes it mandatory for breeders, suppliers and users of laboratory animals to have an animal welfare body (AWB) to prioritize animal welfare and harmonize experimental standards while reassuring the public that research is being carried out appropriately. Based on application of the three Rs (refinement, reduction and replacement), these bodies provide staff with oversight and advisory functions to support compliance with the legal requirements on both animal housing and project realization. This review aims to present the legal measures protecting research animals, with a focus on European AWBs. The review explains how the mission of AWBs includes development of environmental enrichment programs and how animal training generates benefits not only for animal welfare but also for the research work environment and research quality. A French case study is conducted to provide the scientific community with an example of an AWB’s functioning and activities, share its achievements and propose some perspectives for the future.
Health, Psycho-Social Factors, and Ageism in Older Adults in Spain during the COVID-19 Pandemic

Older adults are a population at risk for COVID-19. This study has two independent objectives: (1) to report the impact of COVID-19, as well as psycho-social responses during lockdown, in a sample of older adults in Spain, and (2) to explore through the review of published surveys what extent ageism has increased at the population level. The first objective was to search through an online questionnaire collecting information about self-reported health, lifestyles, psycho-social conditions, and a diversity of concerns. This questionnaire was administered to a volunteer sample of 315 older Spanish men and women (301 COVID-19-free and 14 diagnosed with COVID-19). All individuals reported that they had information about the COVID-19 pandemic. Their self-perception of health was also quite good, most maintaining healthy lifestyles and few reporting unhealthy behaviors. They reported few changes in family and interpersonal relations during lockdown. Those diagnosed with COVID-19 reported higher levels of anxiety, irritation, and fear, in comparison with the COVID-19-free group. Interestingly, instead of being concerned about health, the greatest concern in both groups (COVID-free and those diagnosed with COVID) were politics and the future. Our second objective, to explore ageism during the first wave of the COVID-19 pandemic, was examined across various surveys conducted in several populations by several authors. Results showed an increase in ageism in Spain. Although some new information about health perception, psychosocial responses, and concerns during this unknown stressful situation was obtained, much more research with representative samples is required in order to arrive at more accurate conclusions.
Dry Formulation of Virus-Like Particles in Electrospun Nanofibers

Biologics can be combined with liquid polymer materials and electrospun to produce a dry nanofibrous scaffold. Unlike spray-drying and freeze-drying, electrospinning minimizes the physiological stress on sensitive materials, and nanofiber mat properties such as hydrophobicity, solubility, and melting temperature can be tuned based on the polymer composition. In this study, we explored the dry formulation of a virus-like particle (VLP) vaccine by electrospinning VLP derived from rabbit hemorrhagic disease virus modified to carry the MHC-I gp100 tumor-associated antigen epitope. VLP were added to a polyvinylpyrrolidone (PVP) solution (15% w/v) followed by electrospinning at 24 kV. Formation of a nanofibrous mat was confirmed by scanning electron microscopy, and the presence of VLP was confirmed by transmission electron microscopy and Western blot. VLP from the nanofibers induced T-cell activation and interferon- (IFN-) γ production in vitro. To confirm in vivo cytotoxicity, Pmel mice treated by injection with gp100 VLP from nanofibers induced a gp100 specific immune response, lysing approximately 65% of gp100-pulsed target cells, comparable to mice vaccinated with gp100 VLP in PBS. VLP from nanofibers also induced an antibody response. This work shows that electrospinning can be used to dry-formulate VLP, preserving both humoral and cell-mediated immunity.
Single-Nucleotide Polymorphisms in Oxidative Stress-Related Genes and the Risk of a Stroke in a Polish Population—A Preliminary Study

The present preliminary case-control study was undertaken to detect the potential association of six single nucleotide polymorphisms (SNPs) in oxidative stress-related genes: SOD2 (c.47T > C; rs4880), CAT (c.-89A > T; rs7943316), GPX4 (c.660T > A; rs713041), NOS1 (g.117803515C > T; rs1879417) and NOS2 (c.1823C > T; rs2297518 and c.-227G > C; rs10459953) and the occurrence of a stroke. The SNPs were determined using the TaqMan® Allelic Discrimination Assay in 107 patients with strokes and 107 age- and sex-matched individuals who had not experienced cerebrovascular accidents. The T alleles of the rs4880 were positively correlated with a stroke (bootstrap OR 1.31; 1.07–1.59 95% CI). In the case of the rs713041, an association with the T allele was found (bootstrap OR 1.36; 1.12–1.67). In addition, the occurrence of a stroke was associated with the presence of the C allele of the rs1879417 (bootstrap OR 1.32; 1.09–1.61). We also found that the C/C genotype and C allele of the rs2297518 increased the risk of a stroke (bootstrap ORs 7.00; 4.34–11.29 and 4.96; 3.88–6.34, respectively). Moreover, the C allele of the rs10459953 was associated with an increased occurrence of this disease (bootstrap OR 1.31; 1.08–1.60). These results indicated that genetics variants in the SOD2, GPX4, NOS1 and NOS2 might be associated with susceptibility to strokes in the Polish population.
Impact of Long-Term Irrigation with Sewage on Heavy Metals in Soils, Crops, and Groundwater - a Case Study in Beijing

The effect of sewage irrigation on the accumulation of heavy metals (HMs) in soil profiles, crops, and, groundwater was investigated by monitoring zones with wastewater for various lengths of time (20, 30, and 40 years) in the southeastern suburbs of Beijing. The non-sewage-irrigated region served as the control. Results show that long-term sewage irrigation increased the soil organic matter content. The zone irrigated for 40 years exhibited the greatest accumulations of Hg, Pb, and Cu in the topsoils (0-30 cm). The Cd, Cu, and Zn enrichment was evident in deeper soil layers (40-70 cm) near the sewage waterway. The transfer factors of the various examined HMs in the crop-soil system ranged from 0.002 to 0.491. The HMs of crop grains and kernels in sewage-irrigated zones did not exceed established limits. Long-term sewage irrigation does not constitute HM pollution in soil and shadow groundwater. However, the monitoring of Hg, Pb, and Cu concentrations should be emphasized in areas that engage in treated sewage irrigation to prevent these HMs from entering the food chain and posing health risks.
Nucleoredoxin Knockdown in SH-SY5Y Cells Promotes Cell Renewal

Nucleoredoxin (NXN) is a redox regulator of Disheveled and thereby of WNT signaling. Deficiency in mice leads to cranial dysmorphisms and defects of heart, brain, and bone, suggesting defects of cell fate determination. We used shRNA-mediated knockdown of NXN in SH-SY5Y neuroblastoma cells to study its impact on neuronal cells. We expected that shNXN cells would easily succumb to redox stress, but there were no differences in viability on stimulation with hydrogen peroxide. Instead, the proliferation of naïve shNXN cells was increased with a higher rate of mitotic cells in cell cycle analyses. In addition, basal respiratory rates were higher, whereas the relative change in oxygen consumption upon mitochondrial stressors was similar to control cells. shNXN cells had an increased expression of redox-sensitive heat shock proteins, Hsc70/HSPA8 and HSP90, and autophagy markers suggested an increase in autophagosome formation upon stimulation with bafilomycin and higher flux under low dose rapamycin. A high rate of self-renewal, autophagy, and upregulation of redox-sensitive chaperones appears to be an attractive anti-aging combination if it were to occur in neurons in vivo for which SH-SY5Y cells are a model.
Generation of Photopolymerized Microparticles Based on PEGDA Using Microfluidic Devices. Part 1. Initial Gelation Time and Mechanical Properties of the Material

Photopolymerized microparticles are made of biocompatible hydrogels like Polyethylene Glycol Diacrylate (PEGDA) by using microfluidic devices are a good option for encapsulation, transport and retention of biological or toxic agents. Due to the different applications of these microparticles, it is important to investigate the formulation and the mechanical properties of the material of which they are made of. Therefore, in the present study, mechanical tests were carried out to determine the swelling, drying, soluble fraction, compression, cross-linking density (Mc) and mesh size (ξ) properties of different hydrogel formulations. Tests provided sufficient data to select the best formulation for the future generation of microparticles using microfluidic devices. The initial gelation times of the hydrogels formulations were estimated for their use in the photopolymerization process inside a microfluidic device. Obtained results showed a close relationship between the amount of PEGDA used in the hydrogel and its mechanical properties as well as its initial gelation time. Consequently, it is of considerable importance to know the mechanical properties of the hydrogels made in this research for their proper manipulation and application. On the other hand, the initial gelation time is crucial in photopolymerizable hydrogels and their use in continuous systems such as microfluidic devices.
Early Defoliation Techniques Enhance Yield Components, Grape and Wine Composition of cv. Trnjak (Vitis vinifera L.) in Dalmatian Hinterland Wine Region

Defoliation and cluster thinning are of practical importance in a control of the grapevine source-sink balance, cluster architecture, microclimate and berry composition. Nevertheless, their effectiveness on wine composition is unexplored. In this work, the impacts of preflowering (T1), after berry set (T2), and veraison defoliation (T3) and cluster thinning (T4), on yield components, grape and wine composition of cv. Trnjak are given. Implemented techniques significantly reduced yield and affected grape and wine components in comparison to untreated control (C). Despite lowest number of clusters ensured by cluster thinning, defoliation at veraison had lowest yield. Defoliations improved cluster architecture parameters. Highest berry per se was in preflowering T1 and lowest at veraison T3 defoliation. Berries of T1 had lowest sugar content (19.47 °Brix) while T3 had highest (22.3 °Brix), and the reverse is seen in total acidity highest in T1 (6.12 g/L) and lowest in T3 (5.01 g/L). Wines of early defoliations (T1 and T2) had lowest alcohol and highest anthocyanin concentration. Both techniques applied at veraison produced wines with lower anthocyanins and flavonols than those obtained without any intervention (C). In conclusion, the early defoliations (T1 and T2) improve yield and wine composition of cv. Trnjak in the Mediterranean region of Croatia.
Nanosized Particles Assembled by a Recombinant Virus Protein Are Able to Encapsulate Negatively Charged Molecules and Structured RNA

RNA-based molecules have recently become hot candidates to be developed into therapeutic agents. However, successful applications of RNA-based therapeutics might require suitable carriers to protect the RNA from enzymatic degradation by ubiquitous RNases in vivo. Because of their better biocompatibility and biodegradability, protein-based nanoparticles are considered to be alternatives to their synthetic polymer-based counterparts for drug delivery. Hepatitis C virus (HCV) core protein has been suggested to be able to self-assemble into nucleocapsid-like particles in vitro. In this study, the genomic RNA-binding domain of HCV core protein consisting of 116 amino acids (p116) was overexpressed with E. coli for investigation. The recombinant p116 was able to assemble into particles with an average diameter of approximately 27 nm, as visualized by electron microscopy and atomic force microscopy. Measurements with fluorescence spectroscopy, flow cytometry, and fluorescence quenching indicated that the p116-assembled nanoparticles were able to encapsulate small anionic molecules and structured RNA. This study demonstrates methods that exploit the self-assembly nature of a virus-derived protein for nanoparticle production. This study also suggests that the virus-derived protein-assembled particles could possibly be developed into potential carriers for anionic molecular drugs and structured RNA-based therapeutics.
Genital Brucella suis Biovar 2 Infection of Wild Boar (Sus scrofa) Hunted in Tuscany (Italy)

Brucellosis is a zoonosis caused by different Brucella species. Wild boar (Sus scrofa) could be infected by some species and represents an important reservoir, especially for B. suis biovar 2. This study aimed to investigate the prevalence of Brucella spp. by serological and molecular assays in wild boar hunted in Tuscany (Italy) during two hunting seasons. From 287 animals, sera, lymph nodes, livers, spleens, and reproductive system organs were collected. Within sera, 16 (5.74%) were positive to both rose bengal test (RBT) and complement fixation test (CFT), with titres ranging from 1:4 to 1:16 (corresponding to 20 and 80 ICFTU/mL, respectively). Brucella spp. DNA was detected in four lymph nodes (1.40%), five epididymides (1.74%), and one fetus pool (2.22%). All positive PCR samples belonged to Brucella suis biovar 2. The results of this investigation confirmed that wild boar represents a host for B. suis biovar. 2 and plays an important role in the epidemiology of brucellosis in central Italy. Additionally, epididymis localization confirms the possible venereal transmission.
Role of Heme-Oxygenase-1 in Biology of Cardiomyocytes Derived from Human Induced Pluripotent Stem Cells

Heme oxygenase-1 (HO-1, encoded by HMOX1) is a cytoprotective enzyme degrading heme into CO, Fe2+, and biliverdin. HO-1 was demonstrated to affect cardiac differentiation of murine pluripotent stem cells (PSCs), regulate the metabolism of murine adult cardiomyocytes, and influence regeneration of infarcted myocardium in mice. However, the enzyme’s effect on human cardiogenesis and human cardiomyocytes’ electromechanical properties has not been described so far. Thus, this study aimed to investigate the role of HO-1 in the differentiation of human induced pluripotent stem cells (hiPSCs) into hiPSC-derived cardiomyocytes (hiPSC-CMs). hiPSCs were generated from human fibroblasts and peripheral blood mononuclear cells using Sendai vectors and subjected to CRISPR/Cas9-mediated HMOX1 knock-out. After confirming lack of HO-1 expression on the protein level, isogenic control and HO-1-deficient hiPSCs were differentiated into hiPSC-CMs. No differences in differentiation efficiency and hiPSC-CMs metabolism were observed in both cell types. The global transcriptomic analysis revealed, on the other hand, alterations in electrophysiological pathways in hiPSC-CMs devoid of HO-1, which also demonstrated increased size. Functional consequences in changes in expression of ion channels genes were then confirmed by patch-clamp analysis. To the best of our knowledge, this is the first report demonstrating the link between HO-1 and electrophysiology in human cardiomyocytes.
Temporal and Spatial Epidemiological Analysis of Peste Des Petits Ruminants Outbreaks from the Past 25 Years in Sheep and Goats and Its Control in India

This study was aimed to understand the temporal and spatial epidemiology of peste des petits ruminants (PPR) in India using national surveillance data available in the National Animal Diseases Referral Expert System (NADRES) along with its control plan undertaken. On analysis of the outbreaks/cases reports in sheep and goats in NADRES database from 1995 to 2019, it was observed that PPR features among the top ten diseases and stands first among viral diseases, and among reported deaths, PPR accounts for 36% of mortality in sheep and goats. PPR outbreaks occur round the year in all the seasons but are encountered most frequently during the lean period especially, in the winter season (January to February) in different regions/zones. The reported outbreaks have been progressively declined in most of the states in India due to the implementation of a mass vaccination strategic program since 2011. On state-wise analysis, the PPR risk-areas showed wide variations with different levels of endemicity. Andhra Pradesh, West Bengal, and Karnataka were the top three outbreaks reported states during 1995–2010, whereas Jharkhand and West Bengal states reported more outbreaks during 2011–2015 and 2016–2019 periods. The temporal and spatial distribution of PPR in India provides valuable information on the hotspot areas/zones to take appropriate policy decisions towards its prevention and control in different regions/zones of India. The study also identifies when and where intensive surveillance and vaccination along with biosecurity measures need to be implemented for the control and eradication of the disease from India in consonance with the PPR Global Control and Eradication Strategy.
Arginine Metabolites as Biomarkers of Myocardial Ischaemia, Assessed with Cardiac Magnetic Resonance Imaging in Chronic Kidney Disease

(1) Background: Cardiovascular disease (CVD) is the major cause of morbidity and mortality in patients with chronic kidney disease (CKD). Myocardial oxygenation and perfusion response to stress, using oxygen-sensitive cardiovascular magnetic resonance (OS-CMR) and stress T1 mapping respectively, are impaired in CKD patients with and without known coronary artery disease (CAD). Endothelial dysfunction, assessed by circulating levels of asymmetric dimethylarginine (ADMA) and homoarginine (HMA), promotes atherosclerosis. We hypothesized that in CKD patients, worsening endothelial dysfunction is associated with worsening myocardial oxygenation and perfusion as assessed by change in OS-CMR signal intensity (Δ OS-CMR SI) and stress T1 (ΔT1) values. (2) Methods: 38 patients with advanced CKD underwent cardiovascular magnetic resonance (CMR) scanning at 3 Tesla. OS-CMR and T1 mapping images were acquired both at rest and after adenosine stress and analyzed semi-quantitatively. Serum ADMA and HMA concentrations were assessed using mass spectrometry. (3) Results: There was no significant correlation between Δ OS-CMR SI and ADMA or HMA. Interestingly, there was a significant negative correlation seen between Δ T1 and ADMA (r = −0.419, p = 0.037, n = 30) but not between Δ T1 and HMA. (4) Conclusions: Stress T1 response is impaired in CKD patients and is independently associated with higher circulating ADMA concentrations.
Emergency Lung Transplantation after COVID-19: Immunopathological Insights on Two Affected Patients

We herein characterize the immunopathological features of two Italian COVID-19 patients who underwent bilateral lung transplantation (bLTx). Removed lungs underwent histopathological evaluation. Gene expression profiling (GEP) for immune-related signatures was performed on lung specimens and SARS-CoV-2-stimulated peripheral blood mononuclear cells (PBMCs). Cytokine levels were measured on lungs, bronchoalveolar lavage fluids and in culture supernatants. Pathological assessment showed extensive lung damage with the pattern of proliferative to fibrotic phases, with diffuse alveolar damage mimicking usual interstitial pneumonia (UIP). Lungs’ GEP revealed overexpression of pathogen recognition receptors, effector cytokines and chemokines, immune activation receptors and of the inflammasome components. Multiplex cytokine analysis confirmed a proinflammatory state, with high levels of monocyte/macrophage chemotactic and activating factors and of IL-6 and TNF-α. A similar profile was observed in SARS-CoV-2-stimulated PBMCs collected 7 days after transplant. The pattern of tissue damage observed in the lungs suggests that this may represent the output of protracted disease, resembling a diffuse UIP-like picture. The molecular immune profiling supports the paradigm of a persistent proinflammatory state and sustained humoral immunity, conditions that are maintained despite the iatrogenic immunosuppression.
EPS364, a Novel Deep-Sea Bacterial Exopolysaccharide, Inhibits Liver Cancer Cell Growth and Adhesion

The prognosis of liver cancer was inferior among tumors. New medicine treatments are urgently needed. In this study, a novel exopolysaccharide EPS364 was purified from Vibrio alginolyticus 364, which was isolated from a deep-sea cold seep of the South China Sea. Further research showed that EPS364 consisted of mannose, glucosamine, gluconic acid, galactosamine and arabinose with a molar ratio of 5:9:3.4:0.5:0.8. The relative molecular weight of EPS364 was 14.8 kDa. Our results further revealed that EPS364 was a β-linked and phosphorylated polysaccharide. Notably, EPS364 exhibited a significant antitumor activity, with inducing apoptosis, dissipation of the mitochondrial membrane potential (MMP) and generation of reactive oxygen species (ROS) in Huh7.5 liver cancer cells. Proteomic and quantitative real-time PCR analyses indicated that EPS364 inhibited cancer cell growth and adhesion via targeting the FGF19-FGFR4 signaling pathway. These findings suggest that EPS364 is a promising antitumor agent for pharmacotherapy.