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The development of transition metal-based catalytic platforms that promote bioorthogonal reactions inside living cells remains a major challenge in chemical biology. This is particularly true for palladium-based catalysts, which are very powerful in organic synthesis but perform poorly in the cellular environment, mainly due to their rapid deactivation. We now demonstrate that grafting Pd(II) complexes into engineered ß-sheets of a model WW domain results in cell-compatible palladominiproteins that effectively catalyze depropargylation reactions inside HeLa cells. The concave shape of the WW domain ß-sheet proved particularly suitable for accommodating the metal center and protecting it from rapid deactivation in the cellular environment. A thorough NMR and computational study confirmed the formation of the metal-stapled peptides and allowed us to propose a three-dimensional structure for this novel metalloprotein motif.
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Postpartum reproductive infections in cows generate significant economic losses. The use of lactic acid bacteria in animal health is an alternative tool to avoid antibiotic therapy in the prevention/treatment of bovine reproductive infections. In previous studies, 6 lactic bacteria from bovine mammary glands and vagina with beneficial, safe and technological characteristics were selected, and included in probiotic/phytobiotic formulas (combined with Malva and Lapacho extracts). In this work, probiotic and phytobiotic formulations were designed and their long-term viability determined. They were administered intravaginally to 30 females pregnant bovine pre and postpartum. The modification of the native microbiota and permanence/colonization of cultivable bacteria was evaluated, and also the safety of the designed products through the application of nutritional, clinical, hematological and biochemical parameters. The microorganisms maintained their viability up to 9 months at refrigeration temperature. The number of cultivable bacteria showed different pattern: total aerobic mesophylls increased slightly in all experimental groups, while Enterobacteriaceae increased after delivery, except in beneficial acid lactic bacteria + vegetable extract cows. Control and vegetable extract females showed the highest numbers of Enterobacteriaceae at the end of the trial (30 days postpartum). The number of lactic acid bacteria increased significantly in all the groups between 15 days pre and postpartum. The different parameters evaluated demonstrate the safety and harmlessness of the designed formulas, without producing local and systemic adverse effects in the cows.
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Peel and seeds are the main byproducts from tomato (Lycopersicon esculentum P. Mill) processing with high concentrations of polyphenols that have been underexploited. Herein, polyphenolic profiles in tomato peel and seeds were elucidated by untargeted liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS) with an LTQ Orbitrap analyzer. Samples from two Spanish regionsâ"Murcia" and "Almería"âwere analyzed to obtain complementary results. 57 compounds were found, mainly phenolic acids and flavonoids, of which eight were identified for the first time in tomato. Polyphenols were more abundant in byproducts from "Murcia" samples than in those from"Almería" samples, where the abundance of compounds like coutaric, caffeic, neochlorogenic, dicaffeoylquinic and ferulic acids, vanillic acid hexoside, catechin, naringenin, prunin, apigenin-O-hexoside, rutin, and rutin-O-pentoside was even much higher in byproducts than that in whole fruits. These results reveal the wide range of polyphenols found in tomato byproducts, with potential applications in pharmaceutical research, food preservation, and cosmetic development, among others.
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Frutas , Polifenoles , Semillas , Solanum lycopersicum , Espectrometría de Masas en Tándem , Solanum lycopersicum/química , Polifenoles/análisis , Polifenoles/química , Semillas/química , Espectrometría de Masas en Tándem/métodos , Frutas/química , Cromatografía Líquida de Alta Presión/métodos , Extractos Vegetales/química , Flavonoides/análisis , Flavonoides/químicaRESUMEN
Teriparatide is an anabolic peptide drug indicated for the treatment of osteoporosis. Recombinant teriparatide was first approved in 2002 and has since been followed by patent-free alternatives under biosimilar or hybrid regulatory application. The aim of this study is to demonstrate the essential similarity between synthetic teriparatide BGW and the reference medicinal product (RMP), and thus to ensure the development of the first generic teriparatide drug. Hence, an extensive side-by-side comparative exercise, focusing on structural and biological activity, was performed using a wide range of state-of-the-art orthogonal methods. Nuclear magnetic resonance (NMR), ion mobility-mass spectrometry (IM-MS), UV, circular dichroism (CD) and Fourier transform infrared (FTIR) demonstrated the structural similarity between teriparatide BGW and the RMP. Comparative cell-based bioassays showed that the synthetic and recombinant peptides have identical behaviors. Teriparatide BGW, as a generic drug, provides an available treatment option for patients with osteoporosis and offers clinical benefits identical to those provided by the RMP.
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One of the most used methods to measure antioxidant capacity in food is the ferric reducing antioxidant power (FRAP) test, which is simple, sensitive, and economical, nevertheless has long analysis times, causing measurement errors due to the instability of the FRAP reagent due to its precipitation sequential injection analysis (SIA) is a flow technique that can correct these disadvantages because it is more quickly. So, a novel FRAP-SIA method was developed to evaluate the antioxidant capacity. The system was optimized using a central composite design for hydrodynamic and chemical factors, resulting in a flow rate of 35 µL s-1, and aspirate volumes of 33 µL-38 µL-33 µL for the sequence (FRAP-Antioxidant-FRAP). FRAP reagent was prepared with an HCl solution at 0.005 mol L-1, improving its stability 24 times, concerning when it is in acetate buffer at pH 3.6. The method showed excellent accuracy (RSD <3%) with a LOD of 1.0 µmol L-1 of Trolox for a linear range of 5-120 µmol L-1. The reaction time was diminished by 96% concerning the FRAP-microplate assay (from 30 min to 1.2 min). The method was applied in beverages and extracts, obtaining recovery values ranging from 91.24 to 114.22%.
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Antioxidantes , Alimentos , Antioxidantes/análisis , Oxidación-Reducción , Indicadores y Reactivos , HierroRESUMEN
A full understanding of protein structure is key to unraveling how these systems work, how mutations affect their function, and discovering new hotspots for drug discovery. Research tackling this field began with the analysis of globular proteins. In recent years, as technology has improved, research efforts have broadened their focus to include the study of multidomain proteins and the analysis of conformational variability, flexibility, and dynamic systems. Here, we have selected five recent examples that integrate complementary structural methods to provide insight into the behavior of modular, flexible, and transient contacts. We also describe the structural application of domains derived from single-chain antibodies, which are instrumental in overcoming the size limitation of cryogenic electron microscopy (cryoEM) studies. As these methods are continuously developed, they will lead to the interrogation of more complex systems, revealing how large signaling and transcriptional machines are assembled in the context of health and disease.
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Biología , Conformación MolecularRESUMEN
BACKGROUND: The use of indocyanine green fluorescence has been shown to be a safe and effective method for improving lymph node retrieval in patients with gastric cancer. However, previous studies have focused on early-stage tumors and/or the Asian population and excluded patients who received neoadjuvant treatment. METHODS: In this study, 142 patients with gastric adenocarcinoma underwent laparoscopic gastrectomy at a Spanish hospital between January 2017 and December 2022. Of these, 42 patients received preoperative indocyanine green injection to guide lymphadenectomy. Their outcomes were compared to a retrospective cohort of 42 patients after 1:1 propensity score matching. RESULTS: The feasibility of indocyanine green lymphatic mapping was 95.5%. No complications associated with indocyanine green injection were observed. The indocyanine green group had a significantly higher number of retrieved lymph nodes than the non-indocyanine green group (32.67 vs 25.14; P = .013). This statistically significant difference was maintained across subgroups of neoadjuvant treatment, non-obese patients, pT0 to 2 stage, and pN0 stage. In 47.6% of patients from the indocyanine green group, lymphadenectomy was extended outside the standard D2 dissection area based on indocyanine green uptake, but none of the retrieved lymph nodes were metastatic. There were no differences in postoperative complications and length of hospital stay between the 2 groups. CONCLUSION: Indocyanine green-guided lymphadenectomy is safe and feasible and increases the number of retrieved lymph nodes compared to conventional lymphatic dissection, as well as in patients receiving neoadjuvant chemotherapy. The use of indocyanine green should be routine if available for guiding lymph node dissection in gastric cancer, regardless of tumor stage or previous neoadjuvant treatment. However, further studies are needed to determine the impact of this technique on disease-free and overall survival.
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Laparoscopía , Neoplasias Gástricas , Humanos , Neoplasias Gástricas/tratamiento farmacológico , Neoplasias Gástricas/cirugía , Terapia Neoadyuvante , Estudios Retrospectivos , Verde de Indocianina , Puntaje de Propensión , Escisión del Ganglio Linfático/métodos , Ganglios Linfáticos/cirugía , Ganglios Linfáticos/patología , Laparoscopía/métodos , Gastrectomía/métodosRESUMEN
Many functional aspects of the protein kinase p38α have been illustrated by more than three hundred structures determined in the presence of reducing agents. These structures correspond to free forms and complexes with activators, substrates, and inhibitors. Here we report the conformation of an oxidized state with an intramolecular disulfide bond between Cys119 and Cys162 that is conserved in vertebrates. The structure of the oxidized state does not affect the conformation of the catalytic site, but alters the docking groove by partially unwinding and displacing the short αD helix due to the movement of Cys119 towards Cys162. The transition between oxidized and reduced conformations provides a mechanism for fine-tuning p38α activity as a function of redox conditions, beyond its activation loop phosphorylation. Moreover, the conformational equilibrium between these redox forms reveals an unexplored cleft for p38α inhibitor design that we describe in detail.
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Proteína Quinasa 14 Activada por Mitógenos , Animales , Conformación Proteica , Proteína Quinasa 14 Activada por Mitógenos/metabolismo , Fosforilación/fisiología , Dominio Catalítico , Oxidación-ReducciónRESUMEN
Graft-versus-host disease (GvHD) is a common and severe complication following allogeneic hematopoietic stem cell transplantation (HSCT). Its prevention and treatment is a major challenge. Ferulic acid (FA) has anti-inflammatory and antioxidant properties that could be attractive in this setting. Our aim was to evaluate a bioactive ingredient derived from wheat bran (WB), selected for its high concentration of FA, in a murine model of GvHD. The ingredient was obtained via a bioprocess involving hydrolysis and spray-drying. GvHD was induced via HSCT between MHC-mismatched mouse strains. FA treatment was administered orally. Survival and disease scores (weight loss, hunching, activity, fur texture, and skin integrity, each scored between 0 and 2 depending on disease severity) were recorded daily, histological evaluation was performed at the end of the experiment, and serum inflammatory cytokines were analyzed on days 9 and 28. Treatment with FA did not protect GvHD mice from death, nor did it diminish GvHD scores. However, histological analysis showed that ulcers with large areas of inflammatory cells, vessels, and keratin were less common in skin samples from FA-treated mice. Areas of intense inflammatory response were also seen in fewer small intestine samples from treated mice. In addition, a slight decrease in INF-γ and TNF-α expression was observed in the serum of treated mice on day 28. The results showed some local effect of the ingredient intervention, but that the dose used may not be sufficient to control or reduce the inflammatory response at the systemic level in mice with GvHD. Higher dosages of FA may have an impact when evaluating the immunomodulatory capabilities of the hydrolyzed WB ingredient. Thus, further experiments and the use of technological strategies that enrich the ingredients in soluble ferulic acid to improve its efficacy in this setting are warranted.
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Enfermedad Injerto contra Huésped , Trasplante de Células Madre Hematopoyéticas , Ratones , Animales , Fibras de la Dieta/farmacología , Fibras de la Dieta/uso terapéutico , Modelos Animales de Enfermedad , Enfermedad Injerto contra Huésped/tratamiento farmacológico , Enfermedad Injerto contra Huésped/prevención & control , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Suplementos DietéticosRESUMEN
(1) Background: COVID-19 has evolved during seven epidemic waves in Spain. Our objective was to describe changes in mortality and severity in our hospitalized patients. (2) Method: This study employed a descriptive, retrospective approach for COVID-19 patients admitted to the Hospital de Fuenlabrada (Madrid, Spain) until 31 December 2022. (3) Results: A total of 5510 admissions for COVID-19 were recorded. The first wave accounted for 1823 (33%) admissions and exhibited the highest proportion of severe patients: 65% with bilateral pneumonia and 83% with oxygen saturation under 94% during admission and elevated levels of CRP, IL-6, and D-dimer. In contrast, the seventh wave had the highest median age (79 years) and comorbidity (Charlson: 2.7), while only 3% of patients had bilateral pneumonia and 3% required intubation. The overall mortality rate was 10.3%. The first wave represented 39% of the total. The variables related to mortality were age (OR: 1.08, 1.07-1.09), cancer (OR: 1.99, 1.53-2.60), dementia (OR: 1.82, 1.20-2.75), the Charlson index (1.38, 1.31-1.47), the need for high-flow oxygen (OR: 6.10, 4.94-7.52), mechanical ventilation (OR: 11.554, 6.996-19.080), and CRP (OR: 1.04, 1.03-1.06). (4) Conclusions: The variables associated with mortality included age, comorbidity, respiratory failure, and inflammation. Differences in the baseline characteristics of admitted patients explained the differences in mortality in each wave. Differences observed between patients admitted in the latest wave and the earlier ones suggest that COVID-19 has evolved into a distinct disease, requiring a distinct approach.
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COVID-19 , Epidemias , Humanos , Anciano , COVID-19/epidemiología , Estudios Retrospectivos , Hospitales , HospitalizaciónRESUMEN
Objective and animals: Acute diarrhea is among the most common causes of veterinary consultations for dogs. A double-blind, placebocontrolled intervention trial was done with 120 puppies with gastroenteritis. These dogs were 1 to 4 mo old, male and female, of various breeds and sizes. Procedure: Dogs were randomly allocated into 2 groups: Those in the treated group (TG) received a multi-strain probiotic with Lactobacillus johnsonii CRL1693, Ligilactobacillus murinus CRL1695, Limosilactobacillus mucosae CRL1696, and Ligilactobacillus salivarius CRL1702 (1 × 109 CFU/mL) daily for 7 d, whereas those in the control group (CG) received a placebo. All puppies received intravenous fluids, an antiparasitic, amoxicillin PO, and enrofloxacin SC. Results: At the start of the trial, the 2 groups were similar. Probiotic administration for 7 d normalized fecal consistency, with 69, 50, and 80% of small, medium, and large puppies in the TG achieving a fecal score of 1 (separate hard lumps) at 7 d, significantly better than puppies in the CG. After 7 d of treatment, most puppies (70%) in the TG had an excellent recovery, whereas in the CG, recoveries were 35.7% "bad" and 30.4% "fair." Therefore, treatment with probiotics hastened recovery (P < 0.0001). At the end of the trial, there was a significant increase of cultivable lactobacilli in the feces of TG puppies, but no significant differences between the 2 groups in numbers of total mesophylls, enterobacteria, or Gram-positive cocci. Total mortality was 5.8%, including 4 puppies from the CG and 3 from the TG. Conclusion: In a randomized, double-blind, placebo-controlled study, puppies with gastroenteritis symptoms receiving a multi-strain probiotic had rapid improvement, implying beneficial effects on the microbiota and its functionality.
Un probiotique multi-souches a favorisé la guérison des chiots de la gastro-entérite dans une étude randomisée, en double aveugle et vérifiée par placebo. Objectif et animaux: La diarrhée aiguë fait partie des causes les plus fréquentes de consultations vétérinaires pour les chiens. Un essai d'intervention en double aveugle et vérifié par placebo a été réalisé avec 120 chiots atteints de gastro-entérite. Ces chiens étaient âgés de 1 à 4 mois, mâles et femelles, de différentes races et tailles. Procédure: Les chiens ont été répartis au hasard en 2 groupes : ceux du groupe traité (TG) ont reçu un probiotique multisouches contenant Lactobacillus johnsonii CRL1693, Ligilactobacillus murinus CRL1695, Limosilactobacillus mucosae CRL1696 et Ligilactobacillus salivarius CRL1702 (1 × 109 UFC/mL) quotidiennement pendant 7 j, tandis que ceux du groupe témoin (CG) ont reçu un placebo. Tous les chiots ont reçu des liquides intraveineux, un antiparasitaire, de l'amoxicilline PO et de l'enrofloxacine SC. Résultats: Au début de l'essai, les 2 groupes étaient similaires. L'administration de probiotiques pour une durée de 7 j a normalisé la consistance fécale, avec 69, 50 et 80 % des chiots petits, moyens et grands dans le TG obtenant un score fécal de 1 (morceaux durs séparés) à 7 jours, ce qui était significativement meilleur que les chiots dans le CG. Après 7 jours de traitement, la plupart des chiots (70 %) dans le TG ont eu une excellente récupération, alors que dans le CG, les récupérations étaient de 35,7 % « mauvaises ¼ et 30,4 % « passables ¼. Par conséquent, le traitement avec des probiotiques a accéléré la récupération (P < 0,0001). À la fin de l'essai, il y avait une augmentation significative des lactobacilles cultivables dans les fèces des chiots TG, mais aucune différence significative entre les 2 groupes en nombre de mésophylles totaux, d'entérobactéries ou de coques à Gram positif. La mortalité totale était de 5,8 %, dont 4 chiots du CG et 3 du TG. Conclusion: Dans une étude randomisée, en double aveugle et vérifiée par placebo, des chiots présentant des symptômes de gastro-entérite recevant un probiotique multi-souches ont présenté une amélioration rapide, impliquant des effets bénéfiques sur le microbiote et sa fonctionnalité.(Traduit par Dr Serge Messier).
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Enfermedades de los Perros , Gastroenteritis , Probióticos , Animales , Masculino , Perros , Femenino , Diarrea/terapia , Diarrea/veterinaria , Heces , Gastroenteritis/terapia , Gastroenteritis/veterinaria , Método Doble Ciego , Probióticos/uso terapéutico , Enfermedades de los Perros/tratamiento farmacológicoRESUMEN
BACKGROUND: The aim of this study was to evaluate the impact of a multifaceted intervention to implement an adapted guideline for the management of depression in primary health care. METHODS: A hybrid trial was carried out to determine the effect of a multicomponent provider-centred intervention to improve the detection and diagnosis of depression in primary care, as part of the guideline implementation process, and to collect information about barriers and facilitators in a real-world context. Before the multicomponent intervention, a descriptive cross-sectional study was performed to assess the population prevalence of depression in the participating health centres and to detect possible differences. Subsequently, a quasi-experimental two-phase study was carried out with a concurrent control group to assess the impact of the multicomponent intervention on the main outcomes (detection of depression, evaluation of its severity and the use of structured methods to support the diagnosis). RESULTS: Nine-hundred seventy-four patients took part in the first phase. According to their clinical records, the prevalence of depression ranged from 7.2% to 7.9%, and there were no significant differences between the health centres scheduled to receive the intervention and those in the control group. In the experimental phase, 797 randomly selected participants received the multicomponent intervention. Adjusted multivariable analysis performed before the implementation revealed no significant differences in depression between the experimental and control groups. However, after the intervention, modest but significant differences were observed, which persisted at 1 year after the intervention. CONCLUSIONS: A multicomponent intervention for the implementation of a clinical guideline for the management of depression in primary care produced improvements in the identification of depression and in the degree of severity recorded.
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Depresión , Atención Primaria de Salud , Humanos , Depresión/diagnóstico , Depresión/terapia , Estudios TransversalesRESUMEN
p38α is a versatile protein kinase that can control numerous processes and plays important roles in the cellular responses to stress. Dysregulation of p38α signaling has been linked to several diseases including inflammation, immune disorders and cancer, suggesting that targeting p38α could be therapeutically beneficial. Over the last two decades, numerous p38α inhibitors have been developed, which showed promising effects in pre-clinical studies but results from clinical trials have been disappointing, fueling the interest in the generation of alternative mechanisms of p38α modulation. Here, we report the in silico identification of compounds that we refer to as non-canonical p38α inhibitors (NC-p38i). By combining biochemical and structural analyses, we show that NC-p38i efficiently inhibit p38α autophosphorylation but weakly affect the activity of the canonical pathway. Our results demonstrate how the structural plasticity of p38α can be leveraged to develop therapeutic opportunities targeting a subset of the functions regulated by this pathway.
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Inflamación , Transducción de Señal , Humanos , FosforilaciónRESUMEN
As a first step for the use of probiotics in a formula for cattle, it is required to have available low-cost culture medium(s) and efficient production conditions for the growth of probiotic bacteria and high production of cell biomass. De Man-Rogosa-Sharpe medium, used frequently for lactic acid bacteria (LAB) contains adequate ingredients for their growth but is very expensive for industrial application. The nutrients required for LAB growth are strain-dependent. In this work, traditional culture media were evaluated omitting and/or modifying ingredients in their composition, as carbon or nitrogen source, on the basis of their low-cost industrial waste, to select those supporting the most efficient growth. The results showed that the formulation of culture media containing fructose (0.5%) and molasses (1.0%) was better for the growth and production of cell biomass for all the strains assayed, except Lactobacillus gasseri CRL1421 growing in 1.5% corn syrup. FM902 yeast extract at concentrations between 1.5% and 2.5% was the most adequate for most of the strains. The LAB grown in the designed media maintained the beneficial properties for which they selected. The use of the culture media designed to produce biomass decrease production costs, which is an important step for the feasible industrial production of probiotic pharmaceuticals.
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Lactobacillales , Probióticos , Bovinos , Animales , Biomasa , Melaza , Medios de Cultivo , FermentaciónRESUMEN
Lactic acid bacteria (LAB) were isolated, identified, and characterized from pig feces at various growth stages and feed rations in order to be used as probiotic feed additives. Lactic acid bacteria numbers ranged from 7.10 ± 1.50 to 9.40 log CFUs/g for growing and lactating pigs, respectively. Isolates (n = 230) were identified by (GTG)5-polymerase chain reaction and partial sequence analysis of 16S rRNA. Major LAB populations were Limosilactobacillus reuteri (49.2%), Pediococcus pentosaceus (20%), Lactobacillus amylovorus (11.4%), and L. johnsonii (8.7%). In-vitro assays were performed, including surface characterization and tolerance to acid and bile salts. Several lactobacilli exhibited hydrophobic and aggregative characteristics and were able to withstand gastrointestinal tract conditions. In addition, lactobacilli showed starch- and phytate-degrading ability, as well as antagonistic activity against Gram-negative pathogens and the production of bacteriocin-like inhibitory substances. When resistance or susceptibility to antibiotics was evaluated, high phenotypic resistance to ampicillin, gentamicin, kanamycin, streptomycin, and tetracycline and susceptibility towards clindamycin and chloramphenicol was observed in the assayed LAB. Genotypic characterization showed that 5 out of 15 resistance genes were identified in lactobacilli; their presence did not correlate with phenotypic traits. Genes erm(B), strA, strB, and aadE conferring resistance to erythromycin and streptomycin were reported among all lactobacilli, whereas tet(M) gene was harbored by L. reuteri and L. amylovorus strains. Based on these results, 6 probiotic LAB strains (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T, and L. johnsonii S92R) can be selected to explore their potential as direct feed additives to promote swine health and replace antibiotics.
Des bactéries lactiques (LAB) ont été isolées, identifiées et caractérisées à partir de matières fécales de porc à différents stades de croissance et de rations alimentaires afin d'être utilisées comme additifs alimentaires probiotiques. Le nombre de bactéries lactiques variait de 7,10 ± 1,50 à 9,40 log UFC/g pour les porcs en croissance et en lactation, respectivement. Les isolats (n = 230) ont été identifiés par réaction d'amplification en chaîne par la (GTG)5-polymérase et analyse partielle de la séquence de l'ARNr 16S. Les principales populations de LAB étaient Limosilactobacillus reuteri (49,2 %), Pediococcus pentosaceus (20 %), Lactobacillus amylovorus (11,4 %) et L. johnsonii (8,7 %). Des tests in vitro ont été effectués, y compris la caractérisation de surface et la tolérance aux acides et aux sels biliaires. Plusieurs lactobacilles présentaient des caractéristiques hydrophobes et agrégatives et étaient capables de résister aux conditions du tractus gastro-intestinal. De plus, les lactobacilles ont montré une capacité de dégradation de l'amidon et des phytates, ainsi qu'une activité antagoniste contre les agents pathogènes à Gram négatif et la production de substances inhibitrices de type bactériocine. Lorsque la résistance ou la sensibilité aux antibiotiques a été évaluée, une résistance phénotypique élevée à l'ampicilline, à la gentamicine, à la kanamycine, à la streptomycine et à la tétracycline et une sensibilité à la clindamycine et au chloramphénicol ont été observées dans les LAB testés. La caractérisation génotypique a montré que cinq gènes de résistance sur 15 ont été identifiés dans les lactobacilles; leur présence n'était pas corrélée aux traits phénotypiques. Les gènes erm(B), strA, strB et aadE conférant une résistance à l'érythromycine et à la streptomycine ont été signalés parmi tous les lactobacilles, tandis que le gène tet(M) était hébergé par les souches L. reuteri et L. amylovorus. Sur la base de ces résultats, six souches probiotiques LAB (L. reuteri F207R/G9R/B66R, L. amylovorus G636T/S244T et L. johnsonii S92R) peuvent être sélectionnées pour explorer leur potentiel en tant qu'additifs alimentaires directs pour promouvoir la santé des porcs et remplacer les antibiotiques.(Traduit par Docteur Serge Messier).
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Lactobacillales , Probióticos , Animales , Porcinos , Femenino , Lactobacillales/genética , ARN Ribosómico 16S/genética , Lactancia , Antibacterianos/farmacología , Lactobacillus/genética , Heces/microbiología , Probióticos/farmacología , EstreptomicinaRESUMEN
The curing process (CP) of Vanilla planifolia pods, which is a long and tedious process, is necessary to obtain the natural vanilla extract. This research evaluated the application of microwave (M) and ultrasound (U) during the "killing" stage of the CP and its effect on vanillin content and ß-glucosidase activity. The pods were immersed in a container with water or with moistened samples for the M treatments. In U treatments, the pods were immersed in an ultrasonic bath. After this stage, the samples were subjected to an additional U treatment. The results show that the application of these technologies significantly improves vanillin yield (p < 0.05) and the curing time is reduced to 20 days. U treatments subjected to additional sonication at 38 °C obtain more than double the yield of vanillin regarding control. The effect of M and U on cell structure damage increases with additional sonication, but at 15 min, ß-glucosidase inactivation decreases the final yield. Disposition of samples in M also affects the final vanillin content. There is no significant correlation between ß-glucosidase and vanillin in the different treatments. The application of M and U with the appropriate parameters reduces the CP time without affecting the compounds of interest.
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Lactic acid bacteria (LAB) selected on the basis of probiotic characteristics were administered to beef feedlot catlle and the effect on body condition/growth and nutritional-metabolic status as well as on E. coli O157:H7 fecal shedding, were investigated. A feeding trials involving 126 steers were used to evaluate the effects of Lactobacillus acidophilus CRL2074, Limosilactobacillus fermentum CRL2085 and Limosilactobacillus mucosae CRL2069 and their combinations (5 different probiotic groups and control) when 107-108 CFU/animal of each probiotic group were in-feed supplemented. Cattle were fed a high energy corn-based diet (16 to 88%) and samples from each animal were taken at 0, 40, 104 and 163 days. In general, animals body condition and sensorium state showed optimal muscle-skeletal development and behavioral adaption to confinement; no nasal/eye discharges and diarrheic feces were observed. The nutritional performance of the steers revealed a steady increase of biometric parameters and weight. Animals supplied with L. mucosae CRL2069 for 104 days reached the maximum mean live weight (343.2 kg), whereas the greatest weight daily gain (1.27 ± 0.16 Kg/day) was obtained when CRL2069 and its combination with L. fermentum CRL2085 (1.26 ± 0.11 kg/day) were administered during the complete fattening cycle. With several exceptions, bovine cattle blood and serum parameters showed values within referential ranges. As a preharvest strategy to reduce Escherichia coli O157:H7 in cattle feces, CRL2085 administered during 40 days decreased pathogen shedding with a reduction of 43% during the feeding period. L. fermentum CRL2085 and L. mucosae CRL2069 show promise for feedlot cattle feeding supplementation to improve metabolic-nutritional status, overall productive performance and to reduce E. coli O157:H7 shedding, thus decreasing contamination chances of meat food products.
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Enfermedades de los Bovinos , Infecciones por Escherichia coli , Probióticos , Bovinos , Animales , Escherichia coli , Alimentación Animal/análisis , Probióticos/farmacología , Suplementos Dietéticos , Heces/microbiología , Enfermedades de los Bovinos/prevención & control , Enfermedades de los Bovinos/microbiología , Recuento de Colonia Microbiana/veterinaria , Infecciones por Escherichia coli/veterinariaRESUMEN
The current article provides information that facilitates early identification of a new form of checking obsessive-compulsive disorder (OCD) detected in physicians during the COVID-19 pandemic. This article describes three cases of professional checking OCD in physicians. Physicians with checking OCD are obsessively concerned about making a mistake that will result in fatal consequences. The most frequent strategies of neutralization include avoiding direct contact with patients by taking sick or vacation leaves; compulsively studying for many hours daily (neglecting other aspects of life); checking the status of their patients by arranging additional follow-up consultations or making phone calls even out of working hours; repeatedly checking the medical history of their patients, and persistently recalling the last appointment. Physicians with check OCD often seek reassurance from their colleagues and consult the scientific literature for information about issues they used to be competent in. These patients may also experience anticipatory anxiety and mental blocks. However, egodystony is milder than in other forms of OCD. The COVID pandemic may have exacerbated these neutralization behaviors, since it has forced physicians to adapt to a new work environment. The recommended treatments (Exposure with Response Prevention Therapy or/and SSRI) provide beneficial effects in a short time.
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A µPAD to determine the total polyphenol content (TPC) by Folin-Ciocalteau (FC) method, and antioxidant capacity by ABTS method, coupling an in-situ pretreatment zone with Carrez reagents is proposed. Conditions of FC, K2S2O8, ABTS reagents concentrations, pH, and incubation were established using 2k factorial designs. The proposed µPAD showed linearities upper than 0.99 in ranges 0.150-2.000 mg L-1 and 0.100-1.000 mg L-1 for FC and ABTS methods, respectively, detection limits were lower than 0.135 mg L-1, precision lower than 13.47 %, and recoveries between 82.27 % and 114.60 %. Ten different food samples were analyzed to demonstrate the applicability of the µPAD, and the results were compared with those obtained by the batch methods, showing no significant differences (p > 0.05). The µPAD in-situ sample treatment allowed removing interferences before the TPC and ABTS detection avoiding overestimation of the results.
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Antioxidantes , Polifenoles , Proyectos de Investigación , BenzotiazolesRESUMEN
AIMS: Upper tract urothelial carcinoma (UTUC) is a rare malignancy with a poor prognosis which occurs sporadically or in few cases results from a genetic disorder called Lynch syndrome. Recently, examination of microsatellite instability (MSI) has gained importance as a biomarker: MSI tumours are associated with a better response to immunomodulative therapies. Limited data are known about the prevalence of MSI in UTUC. New detection methods using the fully automated Idylla MSI Assay facilitate analysis of increased patient numbers. METHODS: We investigated the frequency of MSI in a multi-institutional cohort of 243 consecutively collected UTUC samples using standard methodology (Bethesda panel), along with immunohistochemistry of mismatch repair (MMR) proteins. The same tumour cohort was retested using the Idylla MSI Assay by Biocartis. RESULTS: Using standard methodology, 230/243 tumours were detected as microsatellite stable (MSS), 4/243 tumours as MSI and 9/243 samples as invalid. In comparison, the Idylla MSI Assay identified four additional tumours as MSS, equalling 234/243 tumours; 4/243 were classified as MSI and only 5/243 cases as invalid. At the immunohistochemical level, MSI results were supported in all available cases with a loss in MMR proteins. The overall concordance between the standard and the Idylla MSI Assay was 98.35%. Time to result differed between 3 hours for Idylla MSI Assay and 2 days with the standard methodology. CONCLUSION: Our data indicate a low incidence rate of MSI tumours in patients with UTUC. Furthermore, our findings highlight that Idylla MSI Assay can be applied as an alternative method of MSI analysis for UTUC.