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Ganoderma lucidum (a mushroom used in traditional Chinese medicine) compounds may attenuate ageing-related physiological changes and restore normal immunity. However, studies on the physiological effects of Ganoderma lucidum dry extract food supplements are few. Therefore, here, we aimed to investigate the effects of Ganoderma lucidum dry extract food supplement on the lymphocyte function of older women. This was a double-blind clinical trial (n 60) with a final 39 older volunteers, divided into two groups Ganoderma lucidum (n 23) and placebo (n 16). The Ganoderma lucidum group received 2000 mg/d of Ganoderma lucidum dry extract for 8 weeks. We used flow cytometry to determine the lymphocyte profile. CD4+ lymphocyte gene expression was evaluated by real-time polymerase chain reaction. We observed that in the Ganoderma lucidum group, concanavalin A stimulation increased lymphocyte proliferation. Further, we observed an increase in expression of Forkhead box P3, transforming growth factor-beta, IL-10, IL-6, retinoic acid receptor-related orphan receptor gamma, GATA-binding protein 3 and interferon gamma genes in the Ganoderma lucidum group. Furthermore, in the Ganoderma lucidum group, ionomycin and phorbol 12-myristate 13-acetate stimulation led to decrease in Th17+ cells and increase in Th2+ cells. Thus, in older women, Ganoderma lucidum regulates T lymphocyte function leading to a predominant anti-inflammatory action but does not induce T lymphocyte proliferation through CD28 signalling pathway.
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This study critically reevaluates reported Biginelli-like reactions using a Kamlet-Abboud-Taft-based solvent effect model. Surprisingly, structural misassignments were discovered in certain multicomponent reactions, leading to the identification of pseudo three-component derivatives instead of the expected MCR adducts. Attempts to replicate literature conditions failed, prompting reconsideration of the described MCRs and proposed mechanisms. Electrospray ionization (tandem) mass spectrometry, NMR, melting points, elemental analyses and single-crystal X-ray analysis exposed inaccuracies in reported MCRs and allowed for the proposition of a complete catalytic cycle. Biological investigations using both pure and "contaminated" derivatives revealed distinctive features in assessed bioassays. A new cellular action mechanism was unveiled for a one obtained pseudo three-component adduct, suggesting similarity with the known dihydropyrimidinone Monastrol as Eg5 inhibitors, disrupting mitosis by forming monoastral mitotic spindles. Docking studies and RMSD analyses supported this hypothesis. The findings described herein underscore the necessity for a critical reexamination and potential corrections of structural assignments in several reports. This work emphasizes the significance of rigorous characterization and critical evaluation in synthetic chemistry, urging a careful reassessment of reported synthesis and biological activities associated with these compounds.
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Solventes , Solventes/química , Humanos , Cinesinas/antagonistas & inhibidores , Cinesinas/metabolismo , Estructura Molecular , Simulación del Acoplamiento Molecular , Cristalografía por Rayos XRESUMEN
A fossil Mycetophilidae from the Aptian Crato Formation-Cretomanota gondwanica gen. nov., sp. nov.-is described, which is the first mycetophilid from the Crato Formation and corresponds to the oldest known fossil leiine and only the second Gondwanan fossil mycetophilid described so far. Cretomanota gondwanica and both species of Alavamanota Blagoderov and Arillo were added as terminals to the data matrix of a general phylogenetic analysis of the Mycetophilidae, and both fit into the Leiinae. Alavamanota is monophyletic, sister to the clade composed by Cretomanota and the extant genus Manota Williston. The biology of the extant members of this fungivorous family corroborates the reconstruction of the Crato palaeoenvironment as including woodlands with humid habitats and microhabitats. The presence of a Cretaceous member of the tribe Manotini at low latitudes in South America reinforces the hypothesis that the clade with all manotines except Leiella Edwards corresponds to a Lower Cretaceous offshoot from a group in southern Gondwana expanding its distribution to more northern areas into the Gondwana and into Laurasia.
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Dípteros , Fósiles , Animales , Filogenia , Dípteros/genética , Nematocera , América del SurRESUMEN
Cocaine addiction is a relapsing disorder with loss of control in limiting drug intake. Considering the involvement of acetylcholine in the neurobiology of the disease, our aim was to evaluate whether cocaine induces plastic changes in the hippocampal cholinergic muscarinic system. Male Swiss-Webster mice received saline or cocaine (ip) three times daily (60-min intervals) either acutely or in an escalating-dose binge paradigm for 14 days. Locomotor activity was measured in all treatment days. Dopaminergic and cholinergic muscarinic receptors (D1R, D2R, M1-M5, mAChRs), choline acetyltransferase (ChAT), vesicular acetylcholine transporter (VAChT) and acetylcholinesterase (AChE) were quantified in the hippocampus by immunoblotting one hour after the last injection (on drug) or after 14 days of abstinence (withdrawal). Escalating-dose group showed cocaine-induced locomotor sensitization from day 2. M3 mAChR and ChAT significantly increased after the on-drug acute binge treatment. Escalating-dose on-drug group showed increased ChAT, M1, M5 mAChR and D2R; and decreased D1R. Acute-binge withdrawal group showed increased VAChT, M2 mAChR, D1R, and D2R; and decreased M1 mAChR. Escalating-dose withdrawal group presented increased D1R and VAChT and decreased M1 mAChR and D2R. Locomotor activity was negatively correlated with M1 mAChR and AChE in on-drug group and positively correlated with VAChT in withdrawal group. M1 mAChR was positively correlated with M2 mAChR and ChAT in on-drug group, whereas ChAT was positively correlated with M5 mAChR in withdrawal group. The results indicate that cocaine induced an increase in the hippocampal cholinergic tone in the presence of the drug, whereas withdrawal causes a resetting in the system.
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Cocaína , Acetilcolinesterasa/metabolismo , Animales , Colina O-Acetiltransferasa/metabolismo , Colinérgicos , Cocaína/toxicidad , Hipocampo/metabolismo , Masculino , Ratones , Receptores Muscarínicos/metabolismoRESUMEN
Alternanthera brasiliana (L.) Kuntze is recognized for its healing properties; however, its therapeutic effects remain unclear. Therefore, our study aimed to elucidate the wound healing activities of A. brasiliana using in vitro and in vivo assays. In vitro assays were used to evaluate the antibacterial, anti-inflammatory, and antioxidant effects of A. brasiliana extract. For the in vivo study, two dorsal excisions were established in Wistar rats using a punch (1.5 cm in diameter), which were topically treated daily with 2% carbopol gel (Ctrl group) or 20% hydroalcoholic plant extract with 2% carbopol gel (A. brasiliana-Ab group). After the 2nd, 7th, 14th, and 21st days, inflammation, oxidative damage, antioxidants, angiogenesis, tissue formation, and re-epithelialization were evaluated. In vitro, Ab reduced nitric oxide, anion superoxide, and pro-inflammatory cytokine production. In vivo, Ab presented lower levels of inflammatory infiltrate, although increased levels of IL-1ß and TGF-ß1 were observed. The plant extract controlled oxidative damage by antioxidants, which favored angiogenesis, collagenesis, and wound re-epithelialization. Thus, the topical application of the hydroalcoholic extract of 20% A. brasiliana was distinguished by its important anti-inflammatory and antioxidant activities both in vivo and in vitro. The plant extract also stimulated angiogenesis and tissue formation, accelerating total re-epithelization, which is promising for wound healing.
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Amaranthaceae/química , Antiinflamatorios/farmacología , Antioxidantes/farmacología , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/aislamiento & purificación , Antioxidantes/aislamiento & purificación , Línea Celular , Inflamación/tratamiento farmacológico , Inflamación/patología , Masculino , Ratones , Estrés Oxidativo/efectos de los fármacos , Células RAW 264.7 , Ratas , Ratas Wistar , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Recent studies have indicated a prominent role of intestinal microbiota in regulation of several physiological aspects of the host including development and activation of the immune system and control of metabolism. In this review, we focused our discussion on bacterial metabolites produced from dietary fiber fermentation called short-chain fatty acids, which act as a link between the microbiota and host cells. Specifically, we described how modifications in their intestinal levels are associated with development of age-related pathologies including metabolic diseases and type 2 diabetes, hypertension, cardiovascular and neurodegenerative diseases. We also highlight their impact on the development of cancer.
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Envejecimiento/metabolismo , Envejecimiento/patología , Enfermedad , Ácidos Grasos Volátiles/metabolismo , Microbioma Gastrointestinal/fisiología , Humanos , Intestinos/microbiologíaRESUMEN
BACKGROUND: The cytogenomic study of repetitive regions is fundamental for the understanding of morphofunctional mechanisms and genome evolution. Passiflora edulis a species of relevant agronomic value, this work had its genome sequenced by next generation sequencing and bioinformatics analysis performed by RepeatExplorer pipeline. The clusters allowed the identification and characterization of repetitive elements (predominant contributors to most plant genomes). The aim of this study was to identify, characterize and map the repetitive DNA of P. edulis, providing important cytogenomic markers, especially sequences associated with the centromere. RESULTS: Three clusters of satellite DNAs (69, 118 and 207) and seven clusters of Long Terminal Repeat (LTR) retrotransposons of the superfamilies Ty1/Copy and Ty3/Gypsy and families Angela, Athila, Chromovirus and Maximus-Sire (6, 11, 36, 43, 86, 94 and 135) were characterized and analyzed. The chromosome mapping of satellite DNAs showed two hybridization sites co-located in the 5S rDNA region (PeSat_1), subterminal hybridizations (PeSat_3) and hybridization in four sites, co-located in the 45S rDNA region (PeSat_2). Most of the retroelements hybridizations showed signals scattered in the chromosomes, diverging in abundance, and only the cluster 6 presented pericentromeric regions marking. No satellite DNAs and retroelement associated with centromere was observed. CONCLUSION: P. edulis has a highly repetitive genome, with the predominance of Ty3/Gypsy LTR retrotransposon. The satellite DNAs and LTR retrotransposon characterized are promising markers for investigation of the evolutionary patterns and genetic distinction of species and hybrids of Passiflora.
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ADN Satélite/genética , Passiflora/genética , Retroelementos/genética , Mapeo Cromosómico , Cromosomas de las Plantas , ADN de Plantas/genética , ADN de Plantas/metabolismo , ADN Satélite/clasificación , Secuenciación de Nucleótidos de Alto Rendimiento , Hibridación Fluorescente in Situ , Filogenia , ARN Ribosómico/genética , ARN Ribosómico 5S/genética , Análisis de Secuencia de ADNRESUMEN
This work describes a simple approach to overcome challenges in emergency toxicological analysis, using liquid-liquid extraction and high-performance liquid chromatography coupled with a diode-array detector (HPLC-DAD). A rapid procedure has been developed, for the extraction and detection of 19 analytes from the following drug classes: analgesics, benzodiazepines, antidepressants, anticonvulsants and drugs of abuse. These substances are relevant in the context of emergency toxicology in Brazil. The method has been validated according to international guidelines by establishing parameters such as lower limit of quantification, sensitivity, linearity, accuracy and precision. The intra and inter-day precision values, at the lowest concentration levels, have always been less than 20% considering its relative standard deviation. As for accuracy values, these have also been satisfactory (above 81.3%). This method was successfully applied in 201 blood samples from patients with suspected poisoning of the Poison Control Center of São Paulo (PCC-SP), Brazil. Finally, the developed method has shown to be relevant for emergency toxicology due to its high sensitivity and it could be also very useful in both fields of clinical and forensic toxicology.
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Cromatografía Líquida de Alta Presión/métodos , Toxicología Forense/métodos , Preparaciones Farmacéuticas/análisis , Centros de Control de Intoxicaciones , Intoxicación/diagnóstico , Adulto , Brasil , Femenino , Humanos , Límite de Detección , Modelos Lineales , Masculino , Reproducibilidad de los ResultadosRESUMEN
AIM: The purpose of this in vitro study was to evaluate the antimicrobial activity of 2% chlorhexidine gel (CHX) as auxiliary chemical substance and intracanal medications on Candida albicans, Enterococcus faecalis, Escherichia coli, and their endotoxins in the root canals. MATERIALS AND METHODS: The study was conducted on 48 single-rooted human teeth divided into four groups (n = 12), according to intracanal medications used: (1) Calcium hydroxide + apyrogenic saline solution (Ca(OH)2 + SS), (2) 20% ginger glycolic extract (GEN), (3) calcium hydroxide + 20% ginger glycolic extract (Ca(OH)2 + GEN), (4) apyrogenic SS (control). Collections were made from the root canal content before preparation (baseline-S1), immediately after instrumentation (S2), 7 days after instrumentation (S3), after 14 days the action of intracanal medication (S4), and 7 days after removal of the intracanal medication (S5). The antimicrobial activity and endotoxin content were analyzed for all collections. The results were statistically analyzed by the Kruskal-Wallis and Dunn tests at a significance level of 5%. RESULTS: After instrumentation with CHX, there was complete elimination of E. coli and C. albicans, except for E. faecalis, which was significantly reduced and then completely eliminated after intracanal medication. There was significant reduction of endotoxin after instrumentation. Comparison of collection after instrumentation and intracanal medication revealed reduction of endotoxins in all groups; this reduction was greater in group Ca(OH)2 followed by the group GEN. CONCLUSION: It was concluded that the instrumentation using CHX and intracanal medication used were able to eliminate the microorganisms from the root canal; the endotoxins were reduced, yet not completely eliminated. CLINICAL SIGNIFICANCE: This study is important and relevant for searching alternatives during endodontic therapy, since it aims to study the effect of Zingiber officinale on microorganisms and endotoxins present in root canals.
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Bacterias/efectos de los fármacos , Hidróxido de Calcio/farmacología , Clorhexidina/farmacología , Cavidad Pulpar/microbiología , Irrigantes del Conducto Radicular/farmacología , Zingiber officinale , Antibacterianos/farmacología , Candida albicans/efectos de los fármacos , Cavidad Pulpar/química , Cavidad Pulpar/efectos de los fármacos , Endotoxinas , Enterococcus faecalis/efectos de los fármacos , Escherichia coli/efectos de los fármacos , Humanos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Preparación del Conducto Radicular/métodosRESUMEN
Sugarcane is the main source of the world's sugar and is becoming increasingly important as a source of biofuel. The highly polyploid and heterozygous nature of the sugarcane genome has meant that characterization of the genome has lagged behind that of other important crops. Here we developed a method using a combination of quantitative PCR with a transposable marker system to score the relative number of alleles with a transposable element (TE) present at a particular locus. We screened two genera closely related to Saccharum (Miscanthus and Erianthus), wild Saccharum, traditional cultivars, and 127 modern cultivars from Brazilian and Australian breeding programmes. We showed how this method could be used in various ways. First, we showed that the method could be extended to be used as part of a genotyping system. Secondly, the history of insertion and timing of the three TEs examined supports our current understanding of the evolution of the Saccharum complex. Thirdly, all three TEs were found in only one of the two main lineages leading to the modern sugarcane cultivars and are therefore the first TEs identified that could potentially be used as markers for Saccharum spontaneum.
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Marcadores Genéticos , Mutagénesis Insercional , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo Genético , Saccharum/genéticaRESUMEN
Morvan syndrome is a rare condition distinguished by hyperactivity within the central, autonomic, and peripheral nervous systems. Due to the limited number of cases, this presents clinical challenges stemming from the scarcity of published literature. We present a successful anesthetic approach for a patient diagnosed with Morvan syndrome scheduled for elective major intra-thoracic surgery to remove metastases from a thymoma. The patient had previously undergone thymectomy, with the syndrome being diagnosed only one year after the surgery. Additionally, we conducted a literature review on the anesthetic management of this condition.
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The species Passiflora alata, P. cincinnata, and P. edulis have great economic value due to the use of their fruits for human consumption. In this study, we compared the repetitive genome fractions of these three species. The compositions of the repetitive DNA of these three species' genomes were analyzed using clustering and identification of the repetitive sequences with RepeatExplorer. It was found that repetitive DNA content represents 74.70%, 66.86%, and 62.24% of the genome of P. alata, P. edulis, and P. cincinnata, respectively. LTR Ty3/Gypsy retrotransposons represent the highest genome proportions in P. alata and P. edulis, while Ty1/Copia comprises the largest proportion of P. cincinnata genome. Chromosomal mapping by Fluorescent In Situ Hybridization (FISH) showed that LTR retrotransposons have a dispersed distribution along chromosomes. The subtelomeric region of chromosomes is where 145 bp satellite DNA is located, suggesting that these elements may play important roles in genome structure and organization in these species. In this work, we obtained the first global characterization of the composition of repetitive DNA in Passiflora, showing that an increase in genome size is related to an increase in repetitive DNA, which represents an important evolutionary route for these species.
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ADN Satélite , Genoma de Planta , Passiflora , Retroelementos , Passiflora/genética , ADN Satélite/genética , Retroelementos/genética , Cromosomas de las Plantas/genética , Elementos Transponibles de ADN/genética , ADN de Plantas/genética , Hibridación Fluorescente in Situ , Mapeo CromosómicoRESUMEN
Hyperbaric oxygen (HBO) therapy is a well-established method for improving tissue oxygenation and is typically used for the treatment of various inflammatory conditions, including infectious diseases. However, its effect on the intestinal mucosa, a microenvironment known to be physiologically hypoxic, remains unclear. Here, we demonstrated that daily treatment with hyperbaric oxygen affects gut microbiome composition, worsening antibiotic-induced dysbiosis. Accordingly, HBO-treated mice were more susceptible to Clostridioides difficile infection (CDI), an enteric pathogen highly associated with antibiotic-induced colitis. These observations were closely linked with a decline in the level of microbiota-derived short-chain fatty acids (SCFAs). Butyrate, a SCFA produced primarily by anaerobic microbial species, mitigated HBO-induced susceptibility to CDI and increased epithelial barrier integrity by improving group 3 innate lymphoid cell (ILC3) responses. Mice displaying tissue-specific deletion of HIF-1 in RORγt-positive cells exhibited no protective effect of butyrate during CDI. In contrast, the reinforcement of HIF-1 signaling in RORγt-positive cells through the conditional deletion of VHL mitigated disease outcome, even after HBO therapy. Taken together, we conclude that HBO induces intestinal dysbiosis and impairs the production of SCFAs affecting the HIF-1α-IL-22 axis in ILC3 and worsening the response of mice to subsequent C. difficile infection.
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Clostridioides difficile , Infecciones por Clostridium , Microbioma Gastrointestinal , Oxigenoterapia Hiperbárica , Ratones , Animales , Miembro 3 del Grupo F de la Subfamilia 1 de Receptores Nucleares , Inmunidad Innata , Oxigenoterapia Hiperbárica/efectos adversos , Interleucina-22 , Disbiosis/terapia , Linfocitos , Butiratos/farmacología , Ácidos Grasos Volátiles/farmacología , Antibacterianos/farmacologíaRESUMEN
Gut dysbiosis is linked to type 1 diabetes mellitus (T1D). Inulin (INU), a prebiotic, modulates the gut microbiota, promoting beneficial bacteria that produce essential short-chain fatty acids for immune regulation. However, how INU affects T1D remains uncertain. Using a streptozotocin-induced (STZ) mouse model, we studied INU's protective effects. Remarkably, STZ + INU mice resisted T1D, with none developing the disease. They had lower blood glucose, reduced pancreatic inflammation, and normalized serum insulin compared with STZ + SD mice. STZ + INU mice also had enhanced mucus production, abundant Bifidobacterium, Clostridium cluster IV, Akkermansia muciniphila, and increased fecal butyrate. In cecal lymph nodes, we observed fewer CD4+Foxp3+ regulatory T cells expressing CCR4 and more Foxp3+CCR4+ cells in pancreatic islets, with higher CCL17 expression. This phenotype was absent in CCR4-deficient mice on INU. INU supplementation effectively protects against experimental T1D by recruiting CCR4+ regulatory T cells via CCL17 into the pancreas and altering the butyrate-producing microbiota.
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Diabetes Mellitus Tipo 1 , Microbioma Gastrointestinal , Islotes Pancreáticos , Ratones , Animales , Inulina/farmacología , Prebióticos , Modelos Animales de Enfermedad , Linfocitos T Reguladores , Butiratos/farmacología , Factores de Transcripción ForkheadRESUMEN
The primary types, secondary types, and some other identified specimens of 407 Neotropical species of Mycetophilidae at the Natural History Museum, London were examined. Notes were made on the condition of the primary types, their labels, and presence of other specimens in the type series. Additional comments are made about types, secondary types and some few other cases worth of note mistakenly determined to be at the NHM. Lectotypes are designated for syntypes of 17 species: Epicypta insipiens (Williston), Epicypta dolosa (Williston), Leia amabilis Williston, Leia concinna(Williston), Leia nitens (Williston), Megalopelma cellularis Edwards, Megalopelma fraudulenta (Williston), Megalopelma platyura Edwards, Mycetophila borgmeieri Edwards, Mycomya meridionalis Johannsen, Monoclona digitata Edwards, Mycomya peruviana Edwards, Neoempheria maculipennis Williston, Procycloneura paranensisEdwards, Stenophragma nigricauda Edwards, Tetragoneura simplex Edwards, Trizygia nitens Edwards. Three speciesLeia biamputata Edwards, Leia fuscicornis Edwards, and Neallodia flavida Edwardspreviously considered subjective junior synonyms in the literature were revalidated. Mycetophila rufoides nom. nov. is proposed for Mycetophila rufaLane (preocc. Macquart 1826). Mycetophila dolosa Williston is transferred to Epicypta, without being assigned to a particular subgenus. The type of Sceptonia paiaguensis Freeman is formally considered lost. Photographs taken of holotypes and lectotypes are included, helping taxonomic documentation and in some extent, species identification. After the nomenclatural acts in this paper, the Natural History Museum, London, now holds holotypes of 292 Neotropical species of the Mycetophilidae, 23 lectotypes, syntypes of 3 species that have syntypes in other collections, paratypes of 81 species that have holotypes in other collections, identified specimens of 5 species with types lost and specimens of three species which fit in other cases.
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Dípteros/clasificación , Estructuras Animales/anatomía & histología , Animales , Dípteros/anatomía & histología , Femenino , Londres , Masculino , MuseosRESUMEN
Here, in an estuarine canal in southeast Brazil, we evaluated the potential for trace metal accumulation of the acanthocephalan parasite Floridosentis mugilis, which infects the fish host Mugil curema. The quantities of the trace metals were quantified using Inductively Coupled Plasma Mass Spectrometry (ICP-MS), which was used to analyze samples of the fish's muscle, intestine, and liver as well as the parasites. The parasites and the host's tissues had significantly different trace metal concentrations, according to our data. Furthermore, some metals have quite high bioconcentration factors, including Al, Cr, Ni, and Cd. We also found that the trace metal concentrations in the parasites were impacted by the sizes of the parasitic infrapopulations, with smaller infrapopulations tending to accumulate more metals. This study shows this acanthocephalan species' effective ability to store metals and is the first to investigate metal accumulation using it as a model.
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Acantocéfalos , Metales Pesados , Parásitos , Smegmamorpha , Contaminantes Químicos del Agua , Animales , Brasil , Monitoreo del Ambiente/métodos , Contaminantes Químicos del Agua/análisis , Acantocéfalos/química , Metales/análisis , Metales Pesados/análisisRESUMEN
Patients with chronic obstructive pulmonary disease (COPD) may experience exacerbations. During severe exacerbations, nutritional and endocrinological comorbidities can play an important role in the clinical and functional aspects of these patients. The aim of this study was to analyse the influence of the presence of diabetes mellitus (DM) and nutritional parameters on the deterioration of symptoms and quality of life during a severe exacerbation in patients with COPD. An observational study was conducted on COPD patients admitted due to an exacerbation. The COPD Assessment Test (CAT) questionnaire was administered, and clinical and functional parameters were compared based on the presence of nutritional and endocrinological alterations. A total of 50 patients were included, of whom 30 (60%) were male. The mean age was 70.5 years (standard deviation (SD) 9.6). The median CAT score during exacerbation was 25 (interquartile range (IQR) 17.5-30), and the baseline score was 13.5 (IQR 7-19), which represented a statistically significant difference (p < 0.001). Patients with iron deficiencies had a lower total CAT score (p = 0.041), specifically for items related to daily activity (p = 0.009) and energy (p = 0.007). Diabetic patients exhibited a greater decline in pulmonary function during exacerbation (p = 0.016), while patients with high thyroid-stimulating hormone (TSH) levels had a shorter hospital stay (p = 0.016). For COPD patients admitted due to an exacerbation, the metabolic assessment is useful and relevant in the clinical set-up, as endocrinological comorbidities negatively affect clinical and functional aspects of these patients.
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BACKGROUND: The continuous proliferation of intestinal stem cells followed by their tightly regulated differentiation to epithelial cells is essential for the maintenance of the gut epithelial barrier and its functions. How these processes are tuned by diet and gut microbiome is an important, but poorly understood question. Dietary soluble fibers, such as inulin, are known for their ability to impact the gut bacterial community and gut epithelium, and their consumption has been usually associated with health improvement in mice and humans. In this study, we tested the hypothesis that inulin consumption modifies the composition of colonic bacteria and this impacts intestinal stem cells functions, thus affecting the epithelial structure. METHODS: Mice were fed with a diet containing 5% of the insoluble fiber cellulose or the same diet enriched with an additional 10% of inulin. Using a combination of histochemistry, host cell transcriptomics, 16S microbiome analysis, germ-free, gnotobiotic, and genetically modified mouse models, we analyzed the impact of inulin intake on the colonic epithelium, intestinal bacteria, and the local immune compartment. RESULTS: We show that the consumption of inulin diet alters the colon epithelium by increasing the proliferation of intestinal stem cells, leading to deeper crypts and longer colons. This effect was dependent on the inulin-altered gut microbiota, as no modulations were observed in animals deprived of microbiota, nor in mice fed cellulose-enriched diets. We also describe the pivotal role of γδ T lymphocytes and IL-22 in this microenvironment, as the inulin diet failed to induce epithelium remodeling in mice lacking this T cell population or cytokine, highlighting their importance in the diet-microbiota-epithelium-immune system crosstalk. CONCLUSION: This study indicates that the intake of inulin affects the activity of intestinal stem cells and drives a homeostatic remodeling of the colon epithelium, an effect that requires the gut microbiota, γδ T cells, and the presence of IL-22. Our study indicates complex cross kingdom and cross cell type interactions involved in the adaptation of the colon epithelium to the luminal environment in steady state. Video Abstract.
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Microbioma Gastrointestinal , Inulina , Humanos , Animales , Ratones , Inulina/farmacología , Dieta , Fibras de la Dieta , Celulosa , Epitelio , Comunicación CelularRESUMEN
BACKGROUND: To contribute to the knowledge of coleopteran cytogenetics, especially with respect to the genomic content of B chromosomes, we analyzed the composition and organization of repetitive DNA sequences in the Coprophanaeus cyanescens karyotype. We used conventional staining and the application of fluorescence in situ hybridization (FISH) mapping using as probes C0t-1 DNA fraction, the 18S and 5S rRNA genes, and the LOA-like non-LTR transposable element (TE). RESULTS: The conventional analysis detected 3 individuals (among 50 analyzed) carrying one small metacentric and mitotically unstable B chromosome. The FISH analysis revealed a pericentromeric block of C0t-1 DNA in the B chromosome but no 18S or 5S rDNA clusters in this extra element. Using the LOA-like TE probe, the FISH analysis revealed large pericentromeric blocks in eight autosomal bivalents and in the B chromosome, and a pericentromeric block extending to the short arm in one autosomal pair. No positive hybridization signal was observed for the LOA-like element in the sex chromosomes. CONCLUSIONS: The results indicate that the origin of the B chromosome is associated with the autosomal elements, as demonstrated by the hybridization with C0t-1 DNA and the LOA-like TE. The present study is the first report on the cytogenetic mapping of a TE in coleopteran chromosomes. These TEs could have been involved in the origin and evolution of the B chromosome in C. cyanescens.