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We investigated the effects of obesity on metabolic, inflammatory, and oxidative stress parameters in the adipose tissue of patients with fatal COVID-19. Postmortem biopsies of subcutaneous adipose tissue were obtained from 25 unvaccinated inpatients who passed from COVID-19, stratified as nonobese (N-OB; body mass index [BMI], 26.5 ± 2.3 kg m-2) or obese (OB BMI 34.2 ± 5.1 kg m-2). Univariate and multivariate analyses revealed that body composition was responsible for most of the variations detected in the metabolome, with greater dispersion observed in the OB group. Fifteen metabolites were major segregation factors. Results from the OB group showed higher levels of creatinine, myo-inositol, O-acetylcholine, and succinate, and lower levels of sarcosine. The N-OB group showed lower levels of glutathione peroxidase activity, as well as higher content of IL-6 and adiponectin. We revealed significant changes in the metabolomic profile of the adipose tissue in fatal COVID-19 cases, with high adiposity playing a key role in these observed variations. These findings highlight the potential involvement of metabolic and inflammatory pathways, possibly dependent on hypoxia, shedding light on the impact of obesity on disease pathogenesis and suggesting avenues for further research and possible therapeutic targets.
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Autopsia , COVID-19 , Metaboloma , Obesidade , Humanos , COVID-19/metabolismo , COVID-19/mortalidade , COVID-19/patologia , COVID-19/virologia , Obesidade/metabolismo , Obesidade/patologia , Masculino , Feminino , Pessoa de Meia-Idade , Estudos Retrospectivos , Idoso , SARS-CoV-2/metabolismo , Tecido Adiposo/metabolismo , Tecido Adiposo/patologia , Metabolômica/métodos , Índice de Massa Corporal , Adulto , Estresse Oxidativo , Interleucina-6/metabolismoRESUMO
Liquid-phase exfoliation (LPE) in aqueous solutions provides a simple, scalable, and green approach to produce 2D materials. By combining atomistic simulations with exfoliation experiments, the interaction between a surfactant and a 2D layer at the molecular scale can be better understood. In this work, two different dyes, corresponding to rhodamine B base (Rbb) and to a phenylboronic acid BODIPY (PBA-BODIPY) derivative, are employed as dispersants to exfoliate graphene and hexagonal boron nitride (hBN) through sonication-assisted LPE. The exfoliated 2D sheets, mostly as few-layers, exhibit good quality and high loading of dyes. Using molecular dynamics (MD) simulations, the binding free energies are calculated and the arrangement of both dyes on the layers are predicted. It has been found that the dyes show a higher affinity toward hBN than graphene, which is consistent with the higher yields of exfoliated hBN. Furthermore, it is demonstrated that the adsorption behavior of Rbb molecules on graphene and hBN is quite different compared to PBA-BODIPY.
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AIM: To evaluate the inflammatory reaction and the ability to induce mineralization activity of a new repair material, NeoPUTTY (NPutty; NuSmile, USA), in comparison with Bio-C Repair (BC; Angelus, Brazil) and MTA Repair HP (MTA HP; Angelus, Brazil). METHODOLOGY: Polyethylene tubes were filled with materials or kept empty (control group, CG) and implanted in subcutaneous tissue of rats for 7, 15, 30, and 60 days (n = 6/group). Capsule thickness, number of inflammatory cells (ICs), fibroblasts, collagen content, and von Kossa analysis were performed. Unstained sections were evaluated under polarized light and by immunohistochemistry for osteocalcin (OCN). Data were submitted to two-way anova followed by Tukey's test (p ≤ .05), except for OCN. OCN data were submitted to Kruskal-Wallis and Dunn and Friedman post hoc tests followed by the Nemenyi test at a significance level of 5%. RESULTS: At 7, 15, and 30 days, thick capsules containing numerous ICs were seen around the materials. At 60 days, a moderate inflammatory reaction was observed for NPutty, BC while MTA HP presented thin capsules with moderate inflammatory cells. In all periods, NPutty specimens contained the highest values of ICs (p < .05). From 7 to 60 days, the number of ICs reduced significantly while an increase in the number of fibroblasts and birefringent collagen content was observed. At 7 and 15 days, no significant difference was observed in the immunoexpression of OCN (p > .05). At 30 and 60 days, NPutty showed the lowest values of OCN (p < .05). At 60 days, a similar immunoexpression was observed for BC and MTA HP (p > .05). In all time intervals, capsules around NPutty, BC, and MTA HP showed von Kossa-positive and birefringent structures. CONCLUSIONS: Despite the greater inflammatory reaction promoted by NeoPutty than BC and MTA HP, the reduction in the thickness of capsules, the increase in the number of fibroblasts, and the reduction in the number of ICs indicate that this bioceramic material is biocompatible Furthermore, NeoPutty presents the ability to induce mineralization activity.
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Materiais Biocompatíveis , Bismuto , Compostos de Cálcio , Teste de Materiais , Silicatos , Animais , Silicatos/farmacologia , Compostos de Cálcio/farmacologia , Ratos , Materiais Biocompatíveis/farmacologia , Ratos Wistar , Óxidos/farmacologia , Combinação de Medicamentos , Masculino , Compostos de Alumínio/farmacologia , Cimentos Dentários/farmacologia , Fibroblastos/efeitos dos fármacos , Colágeno/metabolismoRESUMO
Carbohydrates and fats constitute our primary energy sources. The importance of each of these energy substrates varies across cell types and physiological conditions. For example, the brain normally relies almost exclusively on glucose oxidation, while skeletal muscle shifts from lipids towards higher carbohydrate oxidation rates as exercise intensity increases. Understanding how carbohydrate are stored in our cells and which tissues contain significant carbohydrate stores is crucial for health professionals, especially given the role of carbohydrate metabolism in various pathophysiological conditions. This laboratory activity uses a simple and low-cost iodine binding method to quantify glycogen in mouse skeletal muscle and liver samples. By integrating the results of this activity with literature data, students can determine overall glycogen storage in the human body. The primary goal of the activity is to enhance students understanding of the importance and limitations of glycogen stores in energy metabolism.
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OBJECTIVE: The cortico-basal ganglia circuit is crucial to understanding locomotor behavior and movement disorders. Spinal cord stimulation modulates that circuit, which is a promising approach to restoring motor functions. However, the effects of electrical spinal cord stimulation in the healthy brain motor circuit in pre- and postgait are poorly understood. Thus, this report aims to evaluate, through electrophysiological analyses, the dynamic spectral features of motor networks underlying locomotor initiation with spinal cord stimulation. MATERIALS AND METHODS: Wistar male rats underwent spinal cord stimulation (current 30-150 µA, frequency 100, 333, and 500 Hz) with the electrophysiological recording of the caudate and putamen nuclei, primary and secondary motor cortices, and primary somatosensory cortex. Video tracking recorded treadmill locomotion and extracted the motor planning and gait initiation. RESULTS: Spectral analysis of segments of gait initiation (pre- and postgait), with stimulation off, showed increased low-frequency activity. Postgait initiation showed increased alpha and beta rhythms and decreased delta rhythm with the stimulation off. Overall, the stimulation frequencies reduced alpha and beta rhythms in all brain areas during movement initiation. Regarding movement planning, such an effect was observed in the sensorimotor area, comprising the delta and alpha rhythms. CONCLUSION: This study showed a short-term effect of spinal cord stimulation on the brain areas of the motor circuit, suggesting possible facilitation of movement planning and starting through neuromodulation. Thus, the electrophysiological characterization of this study may contribute to understanding basal ganglia networks and developing new approaches to treat movement disorders in the gait initiation phase.
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Mites are considered a threat to the commercial poultry industry due to their direct and indirect pathogenic effects. Dermanyssus gallinae is among the mite species that endanger production, as it is hematophagous and poses a serious sanitary risk. Recently, Tyrophagus putrescentiae has also drawn attention as a potential pest in the poultry industry, and its attacks have been related to clinical symptoms such as cutaneous eruptions and bird restlessness. Considering the potential economic, sanitary, and productive losses both species represent to the laying-hen industry, the aim of the present study was to evaluate the influence of D. gallinae and T. putrescentiae on the health and productivity of commercial laying hens by carrying out infestation experiments and by monitoring laying hens for a period of 60 days. Tyrophagus putrescentiae and D. gallinae affected egg quality and zootechnical parameters in laying hens. Hens infested by T. putrescentiae had behavioral changes, weight loss, stress symptoms, and decreased egg laying and quality, as well as diarrhea, gradual feather loss, and itching. Hens infested by D. gallinae also had weight loss, behavioral changes, and stress symptoms, leading to decreased laying and egg quality. The present study proposes an experimental model to understand the impacts caused by T. putrescentiae and D. gallinae on commercial laying production and egg quality parameters.
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Acaridae , Infestações por Ácaros , Ácaros , Doenças das Aves Domésticas , Animais , Feminino , Infestações por Ácaros/veterinária , Galinhas , Redução de PesoRESUMO
Alcohol (ethanol) dependence and related disorders are life-threatening conditions and source of suffering for the user, family members and society. Alcohol withdrawal syndrome (AWS) is a little-known dynamic process associated with a high frequency of relapses. A state of hyperglutamatergic neurotransmission and imbalanced GABAergic function is related to an increased susceptibility to seizures during alcohol withdrawal. Adenosine signaling display an important role in endogenous response to decrease seizure and related damages. Here, an intermittent alcohol exposure regimen (1 h daily of 0.5% ethanol solution) for 16 days or 8 days of the same ethanol exposure regimen followed by 1 or 8 days of ethanol withdrawal was used to assess adenosine signaling in the context of seizure susceptibility using adult zebrafish. In both abstainer groups, a sub-convulsant dose of pentylenetetrazol (2.5 mM) was able to increase the frequency of animals reaching a clonic seizure-like state, while continuous-treated animals had no seizure, as did control animals. The total brain mRNA expression of A1 adenosine receptor was decreased in animals with 1 day of ethanol withdrawal. The agonism of A1 adenosine receptor induced an anticonvulsant effect in animals with 1 day of ethanol withdrawal after the injection of the specific agonist (N6-cyclopentyladenosine, 10 mg.Kg- 1; i.p.). These findings reinforce A1 adenosine receptor as a key target in acute alcohol withdrawal syndrome and zebrafish as an excellent platform to study biological mechanism of AWS.
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Alcoolismo , Síndrome de Abstinência a Substâncias , Animais , Síndrome de Abstinência a Substâncias/tratamento farmacológico , Alcoolismo/tratamento farmacológico , Adenosina/farmacologia , Peixe-Zebra/metabolismo , Anticonvulsivantes/uso terapêutico , Etanol/toxicidade , Convulsões/induzido quimicamente , Convulsões/tratamento farmacológico , Receptores Purinérgicos P1RESUMO
Maple syrup urine disease (MSUD) is an inherited metabolic disorder caused by a deficiency in branched-chain alpha-ketoacid dehydrogenase complex (BCKAC). The treatment is a standard therapy based on a protein-restricted diet with low branched-chain amino acids (BCAA) content to reduce plasma levels and, consequently, the effects of accumulating their metabolites, mainly in the central nervous system. Although the benefits of dietary therapy for MSUD are undeniable, natural protein restriction may increase the risk of nutritional deficiencies, resulting in a low total antioxidant status that can predispose and contribute to oxidative stress. As MSUD is related to redox and energy imbalance, melatonin can be an important adjuvant treatment. Melatonin directly scavenges the hydroxy radical, peroxyl radical, nitrite anion, and singlet oxygen and indirectly induces antioxidant enzyme production. Therefore, this study assesses the role of melatonin treatment on oxidative stress in brain tissue and behavior parameters of zebrafish (Danio rerio) exposed to two concentrations of leucine-induced MSUD: leucine 2 mM and 5mM; and treated with 100 nM of melatonin. Oxidative stress was assessed through oxidative damage (TBARS, DCF, and sulfhydryl content) and antioxidant enzyme activity (SOD and CAT). Melatonin treatment improved redox imbalance with reduced TBARS levels, increased SOD activity, and normalized CAT activity to baseline. Behavior was analyzed with novel object recognition test. Animals exposed to leucine improved object recognition due to melatonin treatment. With the above, we can suggest that melatonin supplementation can protect neurologic oxidative stress, protecting leucine-induced behavior alterations such as memory impairment.
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Doença da Urina de Xarope de Bordo , Melatonina , Animais , Leucina/efeitos adversos , Leucina/metabolismo , Doença da Urina de Xarope de Bordo/metabolismo , Peixe-Zebra/metabolismo , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Melatonina/farmacologia , Melatonina/uso terapêutico , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Estresse Oxidativo , Aminoácidos de Cadeia Ramificada/metabolismo , Superóxido Dismutase/metabolismoRESUMO
Lactation diets dependent on rumen undegradable protein (RUP) sources derived from soybean meal (SBM) products are generally high in Lys and poor in Met. We conducted an experiment to evaluate the effects of increasing dietary RUP and altering digestible AA supply by inclusion of heat-treated soybean meal (HTSBM) or high-protein corn dried distillers grains with soluble (DDGS) on performance in mid-lactation dairy cows. Twenty-four Holstein cows (200 ± 40 d in milk and 30.0 ± 3.92 kg/d of milk yield) blocked according to parity, milk yield, and days in milk were used in a 3 × 3 Latin square design experiment with 21-d periods. Treatments were (1) control (CON), a diet with 6.0% RUP containing 15.9% SBM as the main protein source; (2) HTSBM, a diet with 6.7% RUP containing 4.4% HTSBM partially replacing SBM; and (3) high-protein DDGS (FP; FlexyPro, SJC Bioenergia), a diet with 6.9% RUP containing 5.34% FP partially replacing SBM and ground corn. Diets had similar crude protein (16.9%) and net energy of lactation. Data were submitted to ANOVA using the mixed procedure of SAS software (SAS Institute Inc.). Treatment differences were evaluated using orthogonal contrasts: (1) increasing RUP (SBM vs. HTSBM + FP) and (2) altering digestible AA supply (HTSBM vs. FP). Cows fed HTSBM and FP had greater intake (values in parentheses represent treatment means of CON, HTSBM, and FP, respectively) of neutral detergent fiber (7.14, 7.35, and 7.69 kg/d), crude protein (4.27, 4.37, and 4.51 kg/d), and ether extract (0.942, 0.968, and 1.04 kg/d) compared with cows fed CON. Feeding FP resulted in greater intake of neutral detergent fiber and ether extract compared with HTSBM. Cows fed HTSBM and FP had lower sorting index for feed particles <4 mm than cows fed CON (1.029, 1.008, and 1.022). Feeding FP resulted in greater intake of feed particles <4 mm compared with HTSBM. Treatments containing HTSBM or FP tended to decrease organic matter digestibility (72.4, 71.2, and 71.1%), but no other effects were detected in digestibility of neutral detergent fiber, crude protein, or ether extract. No evidence for differences among treatments was detected in excretion of purine derivatives in milk and urine. Milk yield was greater in cows fed HTSBM or FP than in cows fed CON (28.0, 28.9, and 28.8 kg/d, respectively). Cows fed HTSBM or FP tended to have greater energy-corrected milk and protein yield compared with those fed CON. Milk protein concentration was greater in DDGS cows than those in the HTSBM group (3.45 and 3.40%, respectively). No differences were detected in milk fat yield and concentration, milk urea nitrogen, feed efficiency, or serum concentrations of urea and glucose. Overall, increasing dietary RUP by feeding HTSBM or FP improved intake of nutrients and milk yield without affecting feed efficiency. Altering digestible AA supply while maintaining similar dietary RUP had negligible effects on performance of cows.
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Ração Animal , Zea mays , Gravidez , Feminino , Bovinos , Animais , Zea mays/metabolismo , Ração Animal/análise , Temperatura Alta , Detergentes/metabolismo , Farinha , Lactação , Rúmen/metabolismo , Dieta/veterinária , Glycine max/metabolismo , Proteínas Alimentares/metabolismo , Nutrientes , Ureia/metabolismo , Éteres/metabolismo , Extratos Vegetais/metabolismoRESUMO
Exogenous enzymes are added to diets to improve nutrient utilization and feed efficiency. A study was conducted to evaluate the effects of dietary exogenous enzyme products with amylolytic (Amaize, Alltech) and proteolytic (Vegpro, Alltech) activity on performance, excretion of purine derivatives, and ruminal fermentation of dairy cows. A total of 24 Holstein cows, 4 of which were ruminally cannulated (161 ± 88 d in milk, 681 ± 96 body weight, and 35.2 ± 5.2 kg/d of milk yield), were blocked by milk yield, days in milk, and body weight, and then distributed in a replicated 4 × 4 Latin square design. Experimental periods lasted 21 d, of which the first 14 d were allowed for treatment adaptation and the last 7 d were used for data collection. Treatments were as follows: (1) control (CON) with no feed additives, (2) amylolytic enzyme product added at 0.5 g/kg diet dry matter (DM; AML), (3) amylolytic enzyme product at 0.5 g/kg of diet DM and proteolytic enzyme product at 0.2 g/kg of diet DM (low level; APL), and (4) amylolytic enzyme products added at 0.5 g/kg diet DM and proteolytic enzyme product at 0.4 g/kg of diet DM (high level; APH). Data were analyzed using the mixed procedure of SAS (version 9.4; SAS Institute Inc.). Differences between treatments were analyzed by orthogonal contrasts: CON versus all enzyme groups (ENZ); AML versus APL+APH; and APL versus APH. Dry matter intake was not affected by treatments. Sorting index for feed particles with size <4 mm was lower for ENZ group than for CON. Total-tract apparent digestibility of DM and nutrients (organic matter, starch, neutral detergent fiber, crude protein, and ether extract) were similar between CON and ENZ. Starch digestibility was greater in cows fed APL and APH treatments (86.3%) compared with those in the AML group (83.6%). Neutral detergent fiber digestibility was greater in APH cows compared with those in the APL group (58.1 and 55.2%, respectively). Ruminal pH and NH3-N concentration were not affected by treatments. Molar percentage of propionate tended to be greater in cows fed ENZ treatments than in those fed CON. Molar percentage of propionate was greater in cows fed AML than those fed the blends of amylase and protease (19.2 and 18.5%, respectively). Purine derivative excretions in urine and milk were similar in cows fed ENZ and CON. Uric acid excretion tended to be greater in cows consuming APL and APH than in those in the AML group. Serum urea N concentration tended to be greater in cows fed ENZ than in those fed CON. Milk yield was greater in cows fed ENZ treatments compared with CON (32.0, 33.1, 33.1, and 33.3 kg/d for CON, AML, APL, and APH, respectively). Fat-corrected milk and lactose yields were higher when feeding ENZ. Feed efficiency tended to be greater in cows fed ENZ than in those fed CON. Feeding ENZ benefited cows' performance, whereas the effects on nutrient digestibility were more pronounced when the combination of amylase and protease was fed at the highest dose.
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Doenças dos Bovinos , Leucemia Mieloide Aguda , Feminino , Bovinos , Animais , Lactação , Peptídeo Hidrolases/metabolismo , Propionatos/metabolismo , Fermentação , Detergentes/metabolismo , Digestão , Leite/metabolismo , Dieta/veterinária , Nutrientes , Amido/metabolismo , Peso Corporal , Amilases/metabolismo , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/veterinária , Rúmen/metabolismo , Ração Animal/análise , Zea mays/metabolismo , Doenças dos Bovinos/metabolismoRESUMO
An experiment was conducted to evaluate the effects of inorganic trace minerals (TM) and reduced levels of TM by using proteinate forms of Co, Zn, Mn, and Cu, and Se-yeast in diets of transition cows on performance, TM concentrations in colostrum, plasma, and liver, blood metabolites, antioxidant status, peripheral neutrophil activity, and oocyte quality. Thirty-two Holstein cows (22 multiparous and 10 primiparous cows) were enrolled in this study from 30 d before the expected calving date to 56 DIM. Cows were blocked according to body condition score, parity, and previous milk yield and randomly assigned to one of the following treatments: control (CON), with TM (Zn, Cu, Mn, and Co) supplied in form of sulfate and Se as sodium selenite to meet or exceed requirement estimates of the National Research Council; and proteinate trace minerals (PTM), with TM supplied bound with AA and peptides at 50% of CON levels and inorganic Se replaced with Se-yeast at 100% of CON level. Treatments were supplied until 56 DIM. Eight cows were removed from the study because of early calving (n = 3) or health issues (n = 5); thus, data of 24 cows (16 multiparous and 8 primiparous cows) were used in the statistical analysis. No differences between treatments were detected on nutrient intake or digestibility. Total excretion of purine derivatives was decreased when feeding PTM during the prepartum period. Feeding reduced levels of TM in proteinate form resulted in greater yield of milk (27.7 and 30.9 kg/d for CON and PTM, respectively) and protein (0.890 and 0.976 kg/d) between wk 5 and 8 of lactation. No treatment differences were detected for feed efficiency, milk somatic cell count, and milk urea nitrogen. Cows fed PTM had lower milk fat concentration during the 56 d of evaluation (4.08 and 3.74% for CON and PTM, respectively). Selenium concentration was greater in colostrum of cows fed PTM compared with CON (48.5 and 71.3 µg/L for CON and PTM, respectively), whereas Zn, Cu, and Mn concentrations were not different. Cows fed PTM showed lower liver Cu concentration compared with CON (51.4 and 73.8, respectively). Plasma concentrations of Mn and Zn were lower, but plasma Se concentration tended to be higher with PTM treatment. Feeding PTM resulted in greater blood concentrations of urea-N (16.6 and 18.2 mg/dL for CON and PTM, respectively) and ß-hydroxybutyrate (0.739 and 0.940 mmol/L). Counts of lymphocytes were higher with PTM but counts of monocytes were lower in complete blood cell count. No differences were observed in serum concentrations of superoxide dismutase and glutathione peroxidase. No differences were detected in phagocytosis and oxidative burst potential of neutrophils after incubation with bacteria. Cows fed PTM had fewer viable oocytes per ovum pick-up in comparison with CON (8.00 and 11.6). Feeding PTM to transition cows may sustain performance without altering neutrophil activity despite some alterations in blood TM concentrations. More studies should be performed to evaluate production and fertility measurements when reducing TM dietary levels by using proteinate forms and Se-yeast with larger number of animals.
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Selênio , Oligoelementos , Gravidez , Feminino , Bovinos , Animais , Oligoelementos/metabolismo , Antioxidantes/metabolismo , Selênio/metabolismo , Saccharomyces cerevisiae/metabolismo , Neutrófilos/metabolismo , Ração Animal/análise , Lactação , Leite/química , Dieta/veterinária , Oócitos , Ureia/metabolismo , Período Pós-Parto , Suplementos NutricionaisRESUMO
OBJECTIVES: This study is to evaluate biocompatibility, bioactive potential, porosity, and dentin/material interface of Bio-C Repair (BIOC-R), MTA Repair HP (MTAHP), and Intermediate Restorative Material (IRM). MATERIALS AND METHODS: Dentin tubes were implanted into subcutaneous of rats for 7, 15, 30, and 60 days. Thickness of capsules, number of inflammatory cells (ICs), interleukin-6 (IL-6), osteocalcin (OCN), and von Kossa were evaluated. Porosity and material/dentin interface voids were also analyzed. Data were submitted to ANOVA and Tukey's tests (p < 0.05). RESULTS: IRM capsules were thicker and contained greater ICs and IL-6-immunopositive cells at 7 and 15 days. BIOC-R capsules exhibited higher thickness and ICs at 7 days and greater IL-6 at 7 and 15 days than MTAHP (p < 0.05). At 30 and 60 days, no significant difference was observed among the groups. OCN-immunopositive cells, von Kossa-positive, and birefringent structures were observed in BIOC-R and MTAHP. MTAHP exhibited higher porosity and interface voids (p < 0.05). CONCLUSIONS: BIOC-R, MTAHP, and IRM are biocompatible. Bioceramics materials demonstrate bioactive potential. MTAHP presented the highest porosity and presence of voids. CLINICAL RELEVANCE: BIOC-R and MTAHP have adequate biological properties. BIOC-R demonstrated lower porosity and presence of voids, which may represent better sealing for its clinical applications.
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Materiais Restauradores do Canal Radicular , Ratos , Animais , Materiais Restauradores do Canal Radicular/química , Porosidade , Cápsulas , Interleucina-6 , Teste de Materiais , Compostos de Cálcio/farmacologia , Compostos de Cálcio/química , Silicatos/farmacologia , Silicatos/química , Dentina , Óxidos/química , Combinação de Medicamentos , Compostos de Alumínio/farmacologia , Compostos de Alumínio/químicaRESUMO
Mites are among the major sources of domestic and occupational allergens worldwide, and continuous exposure to these allergens leads to chronic airway inflammation. One of the most allergenic species is the storage mite Tyrophagus putrescentiae (Schrank). Protein extracts are produced from this mite for tests that help the clinical diagnosis (via prick test), treatment, and monitoring of disease progression in patients who had positive results for allergic reactions. Therefore, the aim of the present study was to evaluate the cell viability of RAW 264.7 and L929 cells when exposed to in-house raw protein extracts of T. putrescentiae compared to a commercial product, as well as quantify TNF-α secretion by RAW 264.7. Additionally, this study quantified the effect of these extracts in IgE secretion in total blood of people affected by this mite. The study found similarity between the in-house extract and the commercial extract as they had equivalent TNF-α secretion. Additionally, viabilities of RAW 264.7 and L929 exposed to the in-house extract were compatible with viabilities of cells exposed to the commercial extract, with no cytotoxicity at the concentrations tested. Results corroborated the hypothesis that the extract produced in-house would be equivalent to the commercial extract in allergic patients when the IgE was quantified. This study is the first to show the cytotoxicity of T. putrescentiae extracts, and to provide a quantitative analysis of TNF-α and IgE.
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Acaridae , Hipersensibilidade , Ácaros , Humanos , Animais , Camundongos , Fator de Necrose Tumoral alfa , Imunoglobulina E , Reações Cruzadas , Alérgenos , Ácaros/metabolismoRESUMO
BACKGROUND/AIMS: Pre-mRNA splicing is an essential step in eukaryotic gene expression regulation. Genes are composed of exons that remain in the mature mRNAs and intervening sequences named introns. Splicing is the removal of introns and ligation of exons in a mature transcript. Splice site or spliceosome component mutations can lead to different diseases, including neurodegenerative diseases and several cancer types. HuR is an RNA-binding protein that preferentially binds to U- and AU-rich elements, usually found at the 3' UTRs of some mRNAs. We previously observed HuR specifically associated with spliceosomes assembled on introns containing miR-18a and miR-19a. miR-18a and miR-19a are components of the intronic miR-17-92 cluster, along with other five miRNAs. This cluster has been reported to regulate proliferation, migration, and angiogenesis in cells. In this context, we reasoned HuR could be controlling the splicing and processing of these miRNAs, leading to altered cellular phenotypes. METHODS: We induced HuR overexpression in BCPAP and HEK-293T and analyzed the expression of miRNAs using qPCR, as well as the phenotypic effects in those cells. Cell counting to analyze cell growth was performed after trypan blue staining. Migration and invasion assays were performed using transwell filters and cells were counted after staining with crystal violet. We knocked down HuR using a specific siRNA and analyzed expression of miRNAs by qPCR, as well as cellular kinetics. RESULTS: Our results revealed HuR is associated with miR-19a in BCPAP and HEK-293T cells. Conversely, silencing HuR led to reduced miR-17-5p and miR-19a in BCPAP cells. Our data support that HuR stimulates the expression of miR-19, which is further processed and capable of finding its target sequence in a reporter plasmid. Cells overexpressing HuR showed increased cellular proliferation, migration, and invasion rates. Notably, under the presence of antimiR-19a, BCPAP-HuR cells showed reduced cell growth. Taken together, these results indicate the molecular alterations observed are associated with upregulation of miR-19a, leading to cellular processes involved in cancer development. CONCLUSION: Our findings propose a connection between HuR, miRNA biogenesis and cellular modifications. HuR stimulates miR-19a and miR-19b expression, which leads to up-regulation of cell proliferation, migration and invasion, promoting cancer development.
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MicroRNAs , Neoplasias da Glândula Tireoide , Proteína Semelhante a ELAV 1/genética , Proteína Semelhante a ELAV 1/metabolismo , Humanos , Cinética , MicroRNAs/metabolismo , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide/genéticaRESUMO
Maple Syrup Urine Disease (MSUD) is a metabolic disorder characterized by high levels in blood and urine of branched-chain amino acids leucine, isoleucine, and valine and their alpha-ketoacids, by a partial or total blockade in the activity of branched-chain complex alpha-keto acids dehydrogenase. The main symptoms in MSUD occur in the central nervous system, including cognitive deficits, locomotor, poor feeding, seizures, psychomotor delay, and mental retardation, but the mechanisms of neurotoxicity and behavior alteration due to this disease are poorly understood, thus this study aimed at showing the effects of leucine exposure on glutamate levels and behavior in zebrafish. For this, we analyzed the behavior using the social preference test and novel object recognition test, moreover, we analyse the glutamate levels and uptake using scintillation and high-performance liquid chromatography methods. Our results demonstrated a decrease in glutamate levels and uptake, accompanied by memory and social impairment. In conclusion, these results suggest that alterations in glutamate levels can be associated with behavior impairment, however, more studies are necessary to understand the mechanisms for brain damage in MSUD.
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Doença da Urina de Xarope de Bordo , Peixe-Zebra , Animais , Leucina , Ácido Glutâmico , Doença da Urina de Xarope de Bordo/metabolismo , Aminoácidos de Cadeia Ramificada/farmacologiaRESUMO
Dry malt extract (DME) has been used in animal nutrition as an alternative source of rapidly fermentable carbohydrate. An experiment was conducted to evaluate the partial replacement of ground corn with DME in diets of dairy cows on apparent digestibility, ruminal fermentation, predicted rumen microbial protein supply, N excretion, serum urea-N concentration, and milk yield and composition. Twenty-eight Holstein cows (35.3 ± 5.88 kg/d milk yield and 148 ± 78 d in milk), 4 of which were rumen cannulated, were blocked according to the presence of rumen cannulas, parity, milk yield, and days in milk and enrolled into a crossover design experiment. Experimental periods lasted 21 d, of which the first 14 d were allowed for treatment adaptation and 7 d were used for data collection and sampling. Treatment sequences were composed of control (CON) or DME from barley (Liotécnica Tecnologia em Alimentos) replacing ground corn at 7.62% diet dry matter (~2 kg/d). Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc.) modeling the fixed effects of treatment, period, and their interaction, in addition to the random effect of animal. Ruminal fermentation data were analyzed as repeated measures including time and its interaction with treatment in the previous model as fixed effects. Treatments did not affect nutrient intake or feed sorting. Dry malt extract increased apparent digestibility of CP. Feeding DME decreased ruminal pH and molar percentage of butyrate and increased molar percentage of acetate. No treatment effects were detected for predicted rumen microbial protein supply or N excretion. Cows fed DME had lower serum urea-N concentration than CON cows. Dry malt extract increased yields of actual milk, 3.5% fat-corrected milk, fat, and protein, and improved feed efficiency (fat-corrected milk ÷ dry matter intake). Cows fed DME had lower milk urea nitrogen content in comparison with CON cows. Dry malt extract can partially replace ground corn in the diet while improving milk yield and feed efficiency.
Assuntos
Hordeum , Leite , Ração Animal/análise , Animais , Bovinos , Dieta/veterinária , Digestão , Feminino , Fermentação , Lactação , Leite/química , Nutrientes , Extratos Vegetais/farmacologia , Rúmen/metabolismo , Ureia/metabolismo , Zea mays/químicaRESUMO
Mastitis affects a high proportion of dairy cows and is still one of the greatest challenges faced by the dairy industry. Staphylococcal bacteria remain the most important cause of mastitis worldwide. We investigated how distinct staphylococcal species evade some critical host defense mechanisms, which may dictate the establishment, severity, and persistence of infection and the outcome of possible therapeutic and prevention interventions. Thus, the present study investigated variations among distinct bovine-associated staphylococci in their capability to resist phagocytosis and to trigger respiratory burst activity of blood and milk polymorphonuclear neutrophil leukocytes (PMNL) in dairy cows. To do so, PMNL of 6 primiparous and 6 multiparous dairy cows were used. A collection of 38 non-aureus staphylococci (NAS) and 12 Staphylococcus aureus were included. The phagocytosis and intracellular reactive oxygen species (ROS) production by blood and milk PMNL were analyzed by flow cytometry. Phagocytosis, by both blood and milk PMNL, did not differ between S. aureus and NAS as a group, although within-NAS species differences were observed. Staphylococcus chromogenes (a so-called milk-adapted NAS species) better resisted phagocytosis by blood PMNL than the so-called environmental (i.e., Staphylococcus fleurettii) and opportunistic (i.e., Staphylococcus haemolyticus) NAS species. Otherwise, S. haemolyticus was better phagocytosed by blood PMNL than S. aureus, S. fleurettii, and S. chromogenes. No influence of the origin of the isolates within the staphylococci species in the resistance to phagocytosis by blood and milk PMNL was found. Overall, both S. aureus and NAS did not inhibit intracellular ROS production in blood and milk PMNL. Non-aureus staphylococci induced fewer ROS by milk PMNL than S. aureus, which was not true for blood PMNL, although species-specific differences in the intensity of ROS production were observed. Staphylococcus chromogenes induced more blood PMNL ROS than S. fleurettii and S. haemolyticus, and as much as S. aureus. Conversely, S. chromogenes induced fewer milk PMNL ROS than S. aureus. The origin of the isolates within the staphylococci species did not affect the ROS production by blood and milk PMNL. In conclusion, our study showed differences in staphylococci species in evading phagocytosis and triggering ROS production, which may explain the ability of some staphylococci species (i.e., S. aureus and S. chromogenes) to cause persistent infection and induce inflammation.
Assuntos
Doenças dos Bovinos , Mastite Bovina , Infecções Estafilocócicas , Animais , Bovinos , Feminino , Glândulas Mamárias Animais , Leite , Neutrófilos , Infecção Persistente/veterinária , Fagocitose , Explosão Respiratória , Infecções Estafilocócicas/veterinária , Staphylococcus aureusRESUMO
The inflammasome complex is a key part of chronic diseases and acute infections, being responsible for cytokine release and cell death mechanism regulation. The SARS-CoV-2 infection is characterized by a dysregulated cytokine release. In this context, the inflammasome complex analysis within SARS-CoV-2 infection may prove beneficial to understand the disease's mechanisms. Post-mortem minimally invasive autopsies were performed in patients who died from COVID-19 (n = 24), and lung samples were compared to a patient control group (n = 11) and an Influenza A virus H1N1 subtype group from the 2009 pandemics (n = 10). Histological analysis was performed using hematoxylin-eosin staining. Immunohistochemical (IHC) staining was performed using monoclonal antibodies against targets: ACE2, TLR4, NF-κB, NLRP-3 (or NALP), IL-1ß, IL-18, ASC, CASP1, CASP9, GSDMD, NOX4, TNF-α. Data obtained from digital analysis underwent appropriate statistical tests. IHC analysis showed biomarkers that indicate inflammasome activation (ACE2; NF-κB; NOX4; ASC) were significantly increased in the COVID-19 group (p < 0.05 for all) and biomarkers that indicate cell pyroptosis and inflammasome derived cytokines such as IL-18 (p < 0.005) and CASP1 were greatly increased (p < 0.0001) even when compared to the H1N1 group. We propose that the SARS-CoV-2 pathogenesis is connected to the inflammasome complex activation. Further studies are still warranted to elucidate the pathophysiology of the disease.
Assuntos
COVID-19 , Vírus da Influenza A Subtipo H1N1 , Humanos , Inflamassomos/metabolismo , SARS-CoV-2 , Interleucina-18 , NF-kappa B/metabolismo , Enzima de Conversão de Angiotensina 2 , Autopsia , Vírus da Influenza A Subtipo H1N1/metabolismo , Caspase 1/metabolismo , Pulmão/metabolismo , Citocinas/metabolismo , Biópsia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismoRESUMO
The hematophagous mite Dermanyssus gallinae poses a serious sanitary problem in the Brazilian laying poultry industry. Its control is typically performed with acaricides, either in powder or liquid form. However, the intensive use of these products has caused populations of this species to develop tolerance and even resistance. The aim of the present study is to evaluate the response of eggs and adults of D. gallinae to products in aqueous suspension according to commercial indication and as per the recommendations of the Brazilian Ministry of Agriculture, Livestock and Supply. The study used four acaricide products (product 1: cypermethrin, chlorpyrifos, and piperonyl butoxide; product 2: alkyl-benzyl-dimethyl ammonium chloride, glutaraldehyde, deltamethrin; product 3: dichlorvos; product 4: fluralaner) tested in vitro using the contact method. Distilled water was used in the control group. The effectiveness of each of the products differed significantly between eggs and adults. Products 2, 3, and 4 caused 100% of adult mortality up to day 5 after start of treatment, product 1 97.5%. The corrected mortality (non-viability) of eggs was 21.4% (product 1) 39.4% (product 2), 47.8% (product 3), and 14.4% (product 4). Although the products evaluated were effective against adults of D. gallinae, their effectiveness against eggs was lower under the same conditions. This finding might be directly related to frequent D. gallinae reinfestations in poultry houses.
Assuntos
Infestações por Ácaros , Ácaros , Doenças das Aves Domésticas , Animais , Galinhas , Infestações por Ácaros/prevenção & controle , Infestações por Ácaros/veterinária , Ácaros/fisiologia , ÁguaRESUMO
BACKGROUND: Inflammatory back pain (IBP) is a major criterion in identifying axial spondyloarthritis. Whether socioeconomic issues impact prevalence of IBP assessed using standardized questionnaires has not been assessed. We determined IBP prevalence and performance of IBP questionnaires in a low-income, low-literacy population. METHODS: Individuals were interviewed in Fortaleza, Brazil, for the prevalence of IBP using Calin's, Berlin, and ASAS IBP questionnaires; monthly family income (US dollars), literacy (>/≤8 school years [SYs]), and smoking habit (present/absent) were registered. RESULTS: Two hundred nineteen individuals were included (mean age, 38.2 ± 12.9 years), 110 (50.2%) men, 58 (26.4%) White, and 38 (17.3%) smokers. Overall, 152 (69.4%) declared Assuntos
Dor nas Costas
, Espondilartrite
, Adulto
, Dor nas Costas/epidemiologia
, Brasil/epidemiologia
, Feminino
, Humanos
, Masculino
, Pessoa de Meia-Idade
, Pobreza
, Prevalência
, Espondilartrite/epidemiologia
, Inquéritos e Questionários