Allosteric regulation and crystallographic fragment screening of SARS-CoV-2 NSP15 endoribonuclease.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). The NSP15 endoribonuclease enzyme, known as NendoU, is highly conserved and plays a critical role in the ability of the virus to evade the immune system. NendoU is a promising target for the development of new antiviral drugs. However, the complexity of the enzyme's structure and kinetics, along with the broad range of recognition sequences and lack of structural complexes, hampers the development of inhibitors. Here, we performed enzymatic characterization of NendoU in its monomeric and hexameric form, showing that hexamers are allosteric enzymes with a positive cooperative index, and with no influence of manganese on enzymatic activity. Through combining cryo-electron microscopy at different pHs, X-ray crystallography and biochemical and structural analysis, we showed that NendoU can shift between open and closed forms, which probably correspond to active and inactive states, respectively. We also explored the possibility of NendoU assembling into larger supramolecular structures and proposed a mechanism for allosteric regulation. In addition, we conducted a large fragment screening campaign against NendoU and identified several new allosteric sites that could be targeted for the development of new inhibitors. Overall, our findings provide insights into the complex structure and function of NendoU and offer new opportunities for the development of inhibitors.

Contrasting the stability, octahedral distortions, and optoelectronic properties of 3D MABX3 and 2D (BA)2(MA)B2X7 (B = Ge, Sn, Pb; X = Cl, Br, I) perovskites.

Efficient surface passivation and toxic lead (Pb) are known obstacles to the photovoltaic application of perovskite-based solar cells. A possible solution for these problems is to use thin-films of two-dimensional (2D) perovskite-based materials and the replacement of Pb with alternative divalent cations (B); however, our atomistic understanding of the differences between (3D) three-dimensional and 2D perovskite-based materials is far from satisfactory. Herein, we report a systematic theoretical investigation based on ab initio density functional theory (DFT) calculations for both 3D MABX3 and the Ruddlesden-Popper 2D (BA)2(MA)B2X7 (B = Ge, Sn, Pb, and X = Cl, Br, I) compounds to investigate the differences (contrasts) in selected physical-chemical properties, i.e., structural parameters, energetic stability, electronic, and optical properties. We found an increased cation/anion charge separation because of the presence of organic spacers, which results in stronger Coulomb interactions in the inorganic framework, and hence, it enhances the cohesive energy (stability) within the inorganic layer. The inorganic layer constitutes the optically active region that contributes to the superior performance of perovskite-based solar cells. We quantified this effect by comparing the average electronic charges at the X sites in both 2D and 3D perovskites. This comparison is then correlated with variations in BX6-octahedron volumes, resulting in a monotonic relation. Moreover, the electronic structure characterization demonstrates that Ge-based systems present weakly sensitive band gaps to dimensionality due to a compensatory effect between Jahn-Teller distortions and quantum confinement. Lead-free GeI-, SnBr-, and SnI-based perovskites have DFT band gaps closer to the optimal value used in photovoltaic applications. Finally, as expected, the 2D systems absorption coefficients show pronounced anisotropy.

Nutrient intake, productive and metabolic parameters of Nellore bulls feed a forage free diet using cottonseed cake as a fiber source.

We hypothesized that cottonseed cake in confined Nellore young bulls' diet as fiber source, could maintain or improve the nutrient intake, and productive and metabolic parameters. It was evaluated the total replacement of whole plant corn silage (WPCS) by cottonseed cake (CSC) as a source of fiber in the diet. A completely randomized design with two treatments and 12 replications was used. The treatments were two experimental diets containing 300 g/kg of WPCS or CSC as roughage. All animals at 0, 15, 30, 60 and 112 days were weighed to monitor productive performance and nutrient intake. Dry matter intakes, organic matter, neutral detergent fiber, physically effective neutral detergent fiber, crude protein and non-fiber carbohydrates of the WPCS treatment cattle were higher (p < 0.05), and the ethereal extract intake of cattle fed CSC diet was higher. The BW of cattle fed WPCS was higher in the two initial periods of confinement (p < 0.05). Animals from both treatments showed linear growth during the confinement period. There was a fiber source × period interaction for total weight gain (TWG), average daily gain (ADG) and feed efficiency (FE) (p < 0.05), with the lowest in the first 15 days and the highest at 30 days, reflecting a compensatory gain in animals fed with CSC. Bulls fed with CSC showed negative TWG and ADG in the first period (p < 0.05), but recovered in the following periods, surpassing that of the SC treatment in the last two periods. There was negative EF of bulls fed with CSC in the first period (P < 0.05), but in the following periods the CSC diet did not differ from the WPCS diet. The animals fed CSC diet showed a higher concentration of cholesterol and glucose in the blood plasma (p < 0.05). The forage-free diet containing cottonseed cake as a source of fiber replacing corn silage promoted better animal performance.

Genotype-first approach to identify associations between CDH1 germline variants and cancer phenotypes: a multicentre study by the European Reference Network on Genetic Tumour Risk Syndromes.

BACKGROUND: Truncating pathogenic or likely pathogenic variants of CDH1 cause hereditary diffuse gastric cancer (HDGC), a tumour risk syndrome that predisposes carrier individuals to diffuse gastric and lobular breast cancer. Rare CDH1 missense variants are often classified as variants of unknown significance. We conducted a genotype-phenotype analysis in families carrying rare CDH1 variants, comparing cancer spectrum in carriers of pathogenic or likely pathogenic variants (PV/LPV; analysed jointly) or missense variants of unknown significance, assessing the frequency of families with lobular breast cancer among PV/LPV carrier families, and testing the performance of lobular breast cancer-expanded criteria for CDH1 testing. METHODS: This genotype-first study used retrospective diagnostic and clinical data from 854 carriers of 398 rare CDH1 variants and 1021 relatives, irrespective of HDGC clinical criteria, from 29 institutions in ten member-countries of the European Reference Network on Tumour Risk Syndromes (ERN GENTURIS). Data were collected from Oct 1, 2018, to Sept 20, 2022. Variants were classified by molecular type and clinical actionability with the American College of Medical Genetics and Association for Molecular Pathology CDH1 guidelines (version 2). Families were categorised by whether they fulfilled the 2015 and 2020 HDGC clinical criteria. Genotype-phenotype associations were analysed by Student's t test, Kruskal-Wallis, χ2, and multivariable logistic regression models. Performance of HDGC clinical criteria sets were assessed with an equivalence test and Youden index, and the areas under the receiver operating characteristic curves were compared by Z test. FINDINGS: From 1971 phenotypes (contributed by 854 probands and 1021 relatives aged 1-93 years), 460 had gastric and breast cancer histology available. CDH1 truncating PV/LPVs occurred in 176 (21%) of 854 families and missense variants of unknown significance in 169 (20%) families. Multivariable logistic regression comparing phenotypes occurring in families carrying PV/LPVs or missense variants of unknown significance showed that lobular breast cancer had the greatest positive association with the presence of PV/LPVs (odds ratio 12·39 [95% CI 2·66-57·74], p=0·0014), followed by diffuse gastric cancer (8·00 [2·18-29·39], p=0·0017) and gastric cancer (7·81 [2·03-29·96], p=0·0027). 136 (77%) of 176 families carrying PV/LPVs fulfilled the 2015 HDGC criteria. Of the remaining 40 (23%) families, who did not fulfil the 2015 criteria, 11 fulfilled the 2020 HDGC criteria, and 18 had lobular breast cancer only or lobular breast cancer and gastric cancer, but did not meet the 2020 criteria. No specific CDH1 variant was found to predispose individuals specifically to lobular breast cancer, although 12 (7%) of 176 PV/LPV carrier families had lobular breast cancer only. Addition of three new lobular breast cancer-centred criteria improved testing sensitivity while retaining high specificity. The probability of finding CDH1 PV/LPVs in patients fulfilling the lobular breast cancer-expanded criteria, compared with the 2020 criteria, increased significantly (AUC 0·92 vs 0·88; Z score 3·54; p=0·0004). INTERPRETATION: CDH1 PV/LPVs were positively associated with HDGC-related phenotypes (lobular breast cancer, diffuse gastric cancer, and gastric cancer), and no evidence for a positive association with these phenotypes was found for CDH1 missense variants of unknown significance. CDH1 PV/LPVs occurred often in families with lobular breast cancer who did not fulfil the 2020 HDGC criteria, supporting the expansion of lobular breast cancer-centred criteria. FUNDING: European Reference Network on Genetic Tumour Risk Syndromes, European Regional Development Fund, Fundação para a Ciência e a Tecnologia (Portugal), Cancer Research UK, and European Union's Horizon 2020 research and innovation programme.

SPECT Imaging of Lysyl Oxidase-like 2 in a Model of Idiopathic Pulmonary Fibrosis.

Noninvasive imaging of idiopathic pulmonary fibrosis (IPF) remains a challenge. The aim of this study was to develop an antibody-based radiotracer targeting Lysyl Oxidase-like 2 (LOXL2), an enzyme involved in the fibrogenesis process, for SPECT/CT imaging of pulmonary fibrosis. The bifunctional chelator DOTAGA-PEG4-NH2 was chemoenzymatically conjugated to the murine antibody AB0023 using microbial transglutaminase, resulting in a degree of labeling (number of chelators per antibody) of 2.3. Biolayer interferometry confirmed that the binding affinity of DOTAGA-AB0023 to LOXL2 was preserved with a dissociation constant of 2.45 ± 0.04 nM. DOTAGA-AB0023 was then labeled with 111In and in vivo experiments were carried out in a mice model of progressive pulmonary fibrosis induced by intratracheal administration of bleomycin. [111In]In-DOTAGA-AB0023 was injected in three groups of mice (control, fibrotic, and treated with nintedanib). SPECT/CT images were recorded over 4 days p.i. and an ex vivo biodistribution study was performed by gamma counting. A significant accumulation of the tracer in the lungs of the fibrotic mice was observed at D18 post-bleomycin. Interestingly, the tracer uptake was found selectively upregulated in fibrotic lesions observed on CT scans. Images of mice that received the antifibrotic drug nintedanib from D8 up to D18 showed a decrease in [111In]In-DOTAGA-AB0023 lung uptake associated with a decrease in pulmonary fibrosis measured by CT scan. In conclusion, we report the first radioimmunotracer targeting the protein LOXL2 for nuclear imaging of IPF. The tracer showed promising results in a preclinical model of bleomycin-induced pulmonary fibrosis, with high lung uptake in fibrotic areas, and accounted for the antifibrotic activity of nintedanib.

Tolerability of endometriosis medical treatment: a comparison between combined hormonal contraceptives and progestins.

Endometriosis is a chronic inflammatory disease that occurs in women of reproductive age. Much of the treatment involves hormone therapy that suppresses the proliferation of endometriosis lesions.Objective To compare discontinuation rates of pharmacological treatment with estrogen-progestins and progestins medications. The secondary objective is to evaluate the main side effects of these drugs in patients with endometriosis.Methods This retrospective study analyzed data from 330 patients who attended the Hospital of the State Public Servant of São Paulo from August 1999 to September 2020 and received pharmacological treatment for endometriosis. The data were obtained by review of the files of medical appointments with specialized staff.Results The median treatment time was 18 months, ranging from 1 to 168 months, and 177 patients interrupted the proposed treatment. The combined contraceptives with estrogens and progestins were significantly linked to treatment interruption, with a relative risk of 1,99 (p = 0,005). The most important side effects that resulted in treatment interruption were pain persistence (p = 0,043), weight gain (p = 0,017) and spotting (p < 0,001).

Preanalytical stability of plasma/serum brain-derived tau.

INTRODUCTION: We investigated the effects of matrix type and reagent batch changes on diagnostic performances and longitudinal trajectories of brain-derived tau (BD-tau). METHODS: We evaluated (i) Cohort 1: paired EDTA plasma and serum from Alzheimer biomarker-positive older adults versus controls (n = 26); and (ii) Cohort 2: n = 79 acute ischemic stroke patients with 265 longitudinal samples across four time points. RESULTS: In Cohort 1, plasma and serum BD-tau were strongly correlated (rho = 0.96, p < 0.0001) with similar diagnostic performances (AUCs >99%) and correlations with CSF total-tau (rho = 0.93-0.94, p < 0.0001). However, absolute concentrations were ∼40% higher in plasma versus serum. In Cohort 2, first and repeated BD-tau measurements showed a near-perfect correlation (rho = 0.96, p < 0.0001), with no significant between-batch concentration differences. In longitudinal analyses, substituting ∼10% of the first-run concentrations for the remeasured values showed overlapping estimated trajectories without significant differences at any time point. DISCUSSION: BD-tau has equivalent diagnostic accuracies, but non-interchangeable absolute concentrations, in plasma versus serum. Furthermore, the analytical robustness is unaffected by batch-to-batch reagent variations. HIGHLIGHTS: Brain-derived tau (BD-tau) is a novel blood-based biomarker that quantifies tau protein of CNS origin. Effects of preanalytical handling procedures on the quality and reproducibility of BD-tau measures are unknown. In two cohorts of n = 105 participants, we compared BD-tau concentrations and diagnostic performances in paired plasma and serum samples, and evaluated impacts of batch-to-batch reagent variations. Paired plasma and serum showed equivalent diagnostic performances to separate amyloid-positive AD from amyloid-negative controls, indicating both can be used independently. Repeated measurements and longitudinal trajectories of plasma BD-tau were unaffected by batch-to-batch reagent variation.

Oleanolic Acid Glycosides from Scabiosa caucasica and Scabiosa ochroleuca: Structural Analysis and Cytotoxicity.

In the field of research on medicinal plants from the Armenian flora, the phytochemical study of two Scabiosa L. species, S. caucasica M. Bieb. and S. ochroleuca L. (Caprifoliaceae), has led to the isolation of five previously undescribed oleanolic acid glycosides from an aqueous-ethanolic extract of the roots: 3-O-α-L-rhamnopyranosyl-(1→3)-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid, 3-O-ß-D-xylopyranosyl-(1→2)-[α-L-rhamnopyranosyl-(1→4)]-ß-D-xylopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester, 3-O-α-L-rhamnopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-glucopyranosyl-(1→4)-ß-D-xylopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyloleanolic acid 28-O-ß-D-glucopyranosyl-(1→6)-ß-D-glucopyranosyl ester. Their full structural elucidation required extensive 1D and 2D NMR experiments, as well as mass spectrometry analysis. For the biological activity of the bidesmosidic saponins and the monodesmosidic saponin, their cytotoxicity on a mouse colon cancer cell line (MC-38) was evaluated.

Prediction models of intake and productive performance of non-castrated Nellore cattle finished in the feedlot system under tropical conditions.

The objective of this study was to understand and predict the intake and performance of Nellore cattle finished in the feedlot. Individual data from 144 non-castrated male Nellore steers finished in the feedlot between the years 2016 and 2020 were used. Descriptive statistical analyses and Pearson's correlation were performed. The outliers were tested by evaluating the studentized residuals in relation to the values predicted by the equations. Residues that were outside the range of - 2.5 to 2.5 were removed. The goodness of fit of the developed equations was evaluated by the coefficients of determination (r2) and root mean square error (RMSE). The mean dry matter intake (DMI) was 10.2 kg/day, neutral detergent fiber intake (NDFI) was 3.4 kg/day, corresponding to 33.3% of DMI, crude protein intake (CPI) was 1.6 kg/day, and total digestible nutrient intake (TDNI) was 7.1 kg/day. The CPI to ADG ratio was 1.3 kg CPI/kg ADG and the TDNI to CPI ratio was 4.5 kg TDNI/kg CPI. The averages of productive performance were 1.3 kg/day for average daily gain (ADG), 152.6 kg for total weight gain (TWG), and 497.8 kg for final body weight (FBW) in average days in the confinement of 115.7 days. The intake measures correlated significantly with the performance measures, except for carcass yield and days in the feedlot. TWG had a high positive correlation with ADG (r = 0.84), while FBW had a positive correlation (r = 0.86) with hot carcass weight (HCW). Measures of intake, performance, and days in the feedlot can be used as predictors of DMI, FBW, HCW, TWG, and ADG. The prediction equations had satisfactory precision and accuracy for non-castrated Nellore cattle finished in feedlot systems under tropical conditions.

Effect of physically effective neutral detergent fiber on nutrient intake and digestibility, ruminal and blood parameters, and ingestive behavior of confined beef cattle.

We aimed to evaluate the effect of levels of physically effective neutral detergent fiber from forage (NDFfor) on confined cattle's metabolism and ingestive behavior. Four crossbred steers (514.0 ± 4.54 kg BW) rumen-cannulated were used. The animals were randomly distributed in a 4×4 Latin square design, and the treatments were diets with 9.5%, 5.5%, 2.5%, and 0.0% NDF from whole plant corn silage. The trial was divided into 4 periods of 21 days. The dry matter, organic matter (OM), crude protein, neutral detergent fiber (NDF), physically effective NDF 8mm (peNDF8mm) and NDF1.18mm intakes, and the digestibility of OM and NDF showed a quadratic behavior. The rumen pH values showed a decreasing linear, and the time at pH below 5.8 increased linearly in the diets with lower NDFfor. The production of volatile fatty acids and the proportions of propionate and butyrate showed an increasing quadratic behavior. On the other hand, the proportion of acetate fitted a decreasing quadratic equation. With the lower participation of forage in the diets, the time spent on rumination activity decreased in a quadratic way, and the time spent in idleness increased in a quadratic way. The corn silage can be reduced to 135 g/kg DM in the diet to obtain a minimum of 5.5% of NDF from roughage.

Sequential use of nutritional additives in diets for finishing Nellore steers in confinement.

The objective was to evaluate the effect of using prebiotics (Saccharomyces cerevisiae boulardii) or Monensin in the confinement initial phase and replacing monensin with probiotics (Bacillus toyonensis) in the final phase. Forty-eight Nellore steers were used, with an initial mean body weight of 356.2 ± 17.98 kg, distributed in a completely randomized design. Two animals per pen were confined in 80 m2 pens. The experiment was divided into two stages. The first phase lasted from day 1 to the 30th day, during which the animals were divided into two groups of 24 animals each. The treatments were the nutritional additives added to the diet: monensin or prebiotics (Saccharomyces cerevisiae boulardii). In the second phase, each group was subdivided into 12 animals by treatment, which received monensin or probiotics (Bacillus toyonensis). Dry matter intake (DMI), animal performance, and economic evaluation of the use of additives were evaluated. There was no additive effect on DMI, average daily gain, and total weight gain of the animals in the first experimental stage (0-30th day). Likewise, in the second stage (31st-100th day), there was no treatment effect for the variables of intake and performance. There was no effect of the use of different nutritional additives on carcass characteristics. The use of prebiotics sequentially to probiotics promoted gross and net yield that was superior to that of the animals that consumed monensin. Yeasts and bacteria respectively in the first and second phases of confinement can replace monensin in confinement diets.

Prediction models of carcass characteristics from non­castrated Nellore cattle finished in the feedlot system under tropical conditions.

Our objective was to use measures of intake and productive performance to adjust prediction models for the carcass traits of non-castrated Nellore cattle finished in a feedlot. Individual data from 168 non-castrated male Nellore steers finished in feedlot between the years 2016-2021 were used. Descriptive statistical analyzes and Pearson correlation coefficients were performed. The outliers were tested by evaluating the studentized residuals in relation to the values predicted by the equations. Residues that were outside the range of -2.5 to 2.5 were removed. The goodness of fit of the developed equations was evaluated by the coefficients of determination (R2) and root mean square error (RMSE). Models for carcass yield, subcutaneous fat thickness, ribeye area, and shear force were adjusted. Means of 53.5% carcass yield, 4.8 mm subcutaneous fat thickness, 73 cm2 loin eye area, and 8.1 kg shear force were observed. The observed average intakes were 9.9 kg/day of dry matter, 3.3 kg/day of neutral detergent fiber content, 1.5 kg/day of crude protein, and 7.1 kg/day of total digestible nutrients. The average confinement time was 113 days, the average total weight gain was 152.2 kg and the average daily gain was 1.35 kg/day. Intake measures significantly correlated with shear force and subcutaneous fat thickness and ribeye area. Carcass yield was significantly correlated with total weight gain, feedlot time, and hot carcass weight. Measures of nutrient intake, performance, and confinement time can be used as predictors of carcass yield, ribeye area, fat thickness, and shear force of non-castrated Nellore cattle finished in a feedlot. The prediction equations for ribeye area, carcass yield, subcutaneous fat thickness, and shear force showed sufficient precision and accuracy for non-castrated Nellore cattle finished in confinement systems under tropical conditions. All equations can be used with caution to estimate carcass traits of cattle finished in a feedlot using measures of intake and productive performance.

Effects of extruded urea levels on the productive performance and carcass and meat characteristics of Nellore cattle.

Because the non-protein nitrogen sources can be an option for a ruminant diet to replace true-protein sources, we hypothesized that using extruded urea in increasing levels by reducing soybean meal in the diet can maintain or improve the productive performance of beef cattle in a feedlot. It was aimed to evaluate the effects of extruded urea levels on the productive performance and carcass and meat characteristics of Nellore steers in feedlot. Twenty-four Nellore steers, with an average age of 22 months and an average initial weight of 333.5 kg, were used. A completely randomized design was used with four treatments: 50, 60, 70, and 80 g/100 kg BW of extruded urea. There was no effect of extruded urea levels on nutrient intake. The final weight and average daily gain were adjusted by a quadratic equation, with maximum estimated values of 479.18 kg and 1.02 kg/day with the inclusion of 59.71, 54.14, and 54.16 g/100 kg BW of extruded urea, respectively. There was no effect (P > 0.05) of extruded urea levels on hot carcass weight (233.3 kg), carcass yield (53.2%), ribeye area (72.7 cm2), and subcutaneous fat thickness (5.3 mm). Likewise, there was no effect (P > 0.05) of extruded urea levels on meat chemical composition, shear force (8.9 KgF), cooking losses (28.2%), pH (5.6), and meat color parameters. We recommended the use of 70 g/100 kg BW of extruded urea in the diet for finishing Nellore steers in confinement.

Mathematical models to predict the lamb weight at birth and at weaning reared in the tropical warm-climate pasture.

Our objective was to evaluate the productive parameters of ewes and their lambs in relation to ewe age and to develop models for predicting lamb weight at birth and weaning in a tropical warm-climate pasture. Individual data were collected from 273 ewes and 273 lambs, between 2013 and 2021. During this period all animals were kept on pastures of Brachiaria brizantha cv. Marandu All lambs in the experiments were supplemented in creep-feeding. A descriptive statistical analysis was performed using the PROC SUMMARY procedure in SAS (SAS University Edition, SAS Institute Inc. Cary, CA, USA). Pearson correlation coefficients between variables were estimated using the PROC CORR procedure in SAS (SAS University Edition, SAS Institute Inc. Cary, CA, USA). Model adjustments and variable selection were performed using PROC REG in SAS (SAS University Edition, SAS Institute Inc. Cary, CA, USA). The STEPWISE option and Mallow's C(p) were used to select the variables included in the equations. Outliers were identified by evaluating the studentized residuals based on the predicted values from the equations. Residual analysis was predicted by regression minus observed values and those that fell outside the range of -2.5 to 2.5 were removed. Several statistics were used to assess the predictability of the equations, including the coefficients of determination (r2) and mean standard error (RMSE). The average ewe age at lambing was 3.4 ± 1.7 years, with an average weight of 56.9 ± 8,9 kg and average body condition score (BCS) of 2,4 ± 0.8 points. The average ewe age at weaning was 51.1 ± 7.9, with average BCS of 1.8 ± 0.8 points. The average lamb at birthing was 3.9 ± 0.9 kg. The average lamb at weaning was 21.0 ± 4.9, with daily gain of 0.2 ± 0.1 kg/day and total gain of 17.1 ± 4.7 kg birth to weaning. The lamb produced by ewe at lambing was 5.3 ± 1.7 kg/ewe. The lamb weaned by ewe at weaning was 28.7 ± 10.8 kg/ewe. The ratios of lamb produced per ewe at birth and at weaning were 0.1 ± 0.03 and 0.6 ± 0.2, respectively. The lamb's birth weight showed a positive linear relationship with the age of the ewe, increasing by 115 g per year of age. The regression equations adjusted for ewe age had maximum points ranging from 4.2 to 5.2, occurring at average age of 4,7 years. The other characteristics showed a quadratic tendency. The results suggest the culling of ewes at five years of age to generate lambs with ideal weight at birth and at weaning raised in warm tropical pastures.

MEX3A regulates Lgr5+ stem cell maintenance in the developing intestinal epithelium.

Intestinal stem cells (ISCs) fuel the lifelong self-renewal of the intestinal tract and are paramount for epithelial repair. In this context, the Wnt pathway component LGR5 is the most consensual ISC marker to date. Still, the effort to better understand ISC identity and regulation remains a challenge. We have generated a Mex3a knockout mouse model and show that this RNA-binding protein is crucial for the maintenance of the Lgr5+ ISC pool, as its absence disrupts epithelial turnover during postnatal development and stereotypical organoid maturation ex vivo. Transcriptomic profiling of intestinal crypts reveals that Mex3a deletion induces the peroxisome proliferator-activated receptor (PPAR) pathway, along with a decrease in Wnt signalling and loss of the Lgr5+ stem cell signature. Furthermore, we identify PPARγ activity as a molecular intermediate of MEX3A-mediated regulation. We also show that high PPARγ signalling impairs Lgr5+ ISC function, thus uncovering a new layer of post-transcriptional regulation that critically contributes to intestinal homeostasis.

Fluid biomarkers in stroke: From animal models to clinical care.

Stroke is a leading cause of death and disability worldwide. Stroke prevention, early diagnosis, and efficient acute treatment are priorities to successfully impact stroke death and disability. Fluid biomarkers may improve stroke differential diagnostic, patient stratification for acute treatment, and post-stroke individualized rehabilitation. In the present work, we characterized the use of stroke animal models in fluid biomarker research through a systematic review of PubMed and Scopus databases, followed by a literature review on the translation to the human stroke care setting and future perspectives in the field. We found increasing numbers of publications but with limited translation to the clinic. Animal studies are very heterogeneous, do not account for several human features present in stroke, and, importantly, only a minority of such studies used human cohorts to validate biomarker findings. Clinical studies have found appealing candidates, both protein and circulating nucleic acids, to contribute to a more personalized stroke care pathway. Still, brain tissue complexity and the fact that different brain pathologies share lesion biomarkers make this task challenging due to biomarker low specificity. Moreover, the study design and lack of validation cohorts may have precluded a formal integration of biomarkers in different steps of stroke diagnosis and treatment. To overcome such issues, recent pivotal studies on biomarker dynamics in individual patients are providing added value to diagnosis and anticipating patients' early prognosis. Presently, the most consistent protein biomarkers for stroke diagnosis and short- and long-term prognosis are associated with tissue damage at neuronal (TAU), axonal (NFL), or astroglial (GFAP and S100ß) levels. Most promising nucleic acids are microRNAs (miR), due to their stability in plasma and ease of access. Still, clinical validation and standardized quantitation place them a step behind compared protein as stroke biomarkers. Ultimately, the definition of clinically relevant biomarker panels and optimization of fast and sensitive biomarker measurements in the blood, together with their combination with clinical and neuroimaging data, will pave the way toward personalized stroke care.

IL-1RAP, a Key Therapeutic Target in Cancer.

Cancer is a major cause of death worldwide and especially in high- and upper-middle-income countries. Despite recent progress in cancer therapies, such as chimeric antigen receptor T (CAR-T) cells or antibody-drug conjugate (ADC), new targets expressed by the tumor cells need to be identified in order to selectively drive these innovative therapies to tumors. In this context, IL-1RAP recently showed great potential to become one of these new targets for cancer therapy. IL-1RAP is highly involved in the inflammation process through the interleukins 1, 33, and 36 (IL-1, IL-33, IL-36) signaling pathways. Inflammation is now recognized as a hallmark of carcinogenesis, suggesting that IL-1RAP could play a role in cancer development and progression. Furthermore, IL-1RAP was found overexpressed on tumor cells from several hematological and solid cancers, thus confirming its potential involvement in carcinogenesis. This review will first describe the structure and genetics of IL-1RAP as well as its role in tumor development. Finally, a focus will be made on the therapies based on IL-1RAP targeting, which are now under preclinical or clinical development.

Characterization and in vitro evaluations of Baru pulp from almond production waste ensiled with different additives.

The silage process is an efficient way of storing nutrients for animal nutrition. Our hypothesis was that the Baru pulp can be preserved in the form of silage, regardless of the use of additives to aid the process. Silages of Baru pulp containing different additives were evaluated through in vitro analysis, fermentation parameters, and composition and thermal analysis including differential scanning calorimetry and thermogravimetry. The treatments consisted of (1) silage with Baru pulp (BP) in natura without additive; (2) BP in natura with acetic acid; (3) BP in natura with formic acids; and (4) BP in natura added with microbial inoculums in a randomized experimental design with three replications per treatment and analyzed in duplicate. The ensiled material was kept in anaerobic conditions for a period of 30 days. BP before and after the silage process presented averages of 67.31 and 66.24% for in vitro digestibility of DM (IVDMD). Microbial inoculant additive was the most effective in reducing pH, followed by acetic acid and formic acid. There were effects of additives on silages for all degradation parameters in ruminal liquid in vitro. It was observed that BP before ensiling had the highest A fraction (7.9 mL gas/100 mg DM), without differing from the silage treated with formic acid (7.1 mL gas/100 mg DM). Similar effects were observed on mass loss (TG) and heat flux (DSC) between the silages. Our findings suggest that Baru pulp with formic acid is more efficient in the conservation and preservation of fermentable carbohydrates as well as in silage production.

Ruminal kinetics and nutritive value of Zuri grass silage harvested at different ages and added with powder molasses.

This study aimed to evaluate the effect of regrowth ages and the use of powdered molasses on nutritional characteristics of Zuri grass (Megathyrsus maximus) silages. A completely randomized design was used, in a 3 × 3 factorial scheme: three regrowth ages (65, 80, and 95 days) and three inclusion levels of powdered molasses of sugarcane (0, 20, and 40 g), with four replications. The chemical composition, fermentation parameters, in vitro digestibility, and in vitro degradation kinetics of the silages were evaluated. Greater dry matter (DM) contents were observed in silages with 95 days with 40 g of molasses. The 65 days of regrowth silages with 40 g of molasses had greater in vitro digestibility of dry matter compared to other treatments. However, lower pH values, N-NH3, DM losses, gas losses, and density were observed in silages with 95 days of regrowth with 40 g of molasses. It is recommended to cut the Zuri grass in the range of 80 to 95 days of regrowth with the addition of 40 g/kg of molasses in powders for the production of silages.

Cotton cake as an economically viable alternative fibre source of forage in a high-concentrate diet for finishing beef cattle in feedlots.

Cotton cake can be a sustainable and economically viable alternative to maize silage as an effective fibre source in the finishing diet of feedlot beef cattle. Thus, the objective was to evaluate the economic viability and the productive performance of Nellore steers finished in feedlots using cotton cake as a source of fibre, replacing maize silage in the diet. Twenty-four non-castrated Nellore steers, with an average weight of 377.8 kg and an age of approximately 24 months, kept in individual pens, were used. The diet was 30% roughage, in the form of maize silage, and a diet without forage containing 30% cotton cake as an effective fibre source, based on dry matter. The design used was completely randomised with two treatments and 12 replications. Effects (p < 0.05) of the dietary fibre source were observed for final body weight (BW), total weight gain, hot carcass weight (HCW) and carcass yield (CY). Revenue and net margin were higher for the treatment with cotton cake. The corn silage diet promoted higher total costs per animal (U$ 408.41 vs. U$ 336.06) and daily costs (U$ 3.65 vs. U$ 3.00). Greater differences were observed between fibre sources for the concentrate cost, and the maize silage diet had a cost U$ 98.29 higher than the treatment with cotton cake (U$ 314.04 vs. U$ 215.75). The cotton cake treatment had a higher cost of U$106.98/animal for the net margin. Cotton cake is a viable alternative and can be used as a source of fibre in forage-free diets without affecting production performance, promoting benefits in economic performance for finishing beef steers in feedlots.