RESUMO
BACKGROUND: Joint flexion to diminish the gap and avoid nerve grafts fell into disuse for decades, but recently attention for using this technique was regained. We report a case series of nerve suture under joint flexion, ultrasound monitoring, and physiotherapy. Our main objective was to determine how effective this multimodality treatment is. METHODS: A retrospective review of 8 patients treated with direct repair with joint flexion was done. Depending on the affected nerve, either the knee or the elbow was flexed intraoperatively to determine if direct suturing was possible. After surgery, the limb was held immobilized. Through serial ultrasounds and a physiotherapy program, the limb was fully extended. If a nerve repair rupture was observed, the patient was re-operated and grafts were used. RESULTS: Of the eight nerve sutures analyzed, four sustained a nerve rupture revealed by US at an early stage, while four did not show any sign of dehiscence. In the patients in whom the nerve suture was preserved, an early and very good response was observed. Ultrasound was 100% accurate at identifying nerve suture preservation. Early detection of nerve failure permitted early re-do surgery using grafts without flexion, ultimately determining good final results. CONCLUSIONS: We observed a high rate of dehiscence in our group of patients treated with direct repair and joint flexion. We believe this was due to an incorrect use of the immobilization device, excessive movement, or a broken device. In opposition to this, we observed that applying direct nerve sutures and joint flexion offers unusually good and fast results. If this technique is employed, it is mandatory to closely monitor suture status with US, together with physiotherapy providing progressive, US-guided extension of the flexed joint. If nerve rupture occurs, the close monitoring dictated by this protocol should ensure the timely application of a successful graft repair.
Assuntos
Procedimentos de Cirurgia Plástica , Suturas , Humanos , Modalidades de Fisioterapia , Amplitude de Movimento Articular , Ruptura/cirurgiaRESUMO
Superficial mycoses are the most frequently diagnosed affections of the stratum corneum of the skin, nails and hair. It is generally caused by the presence of yeasts and dermatophytes. Onychomycosis is the most common infection with an incidence of 80-90% in Europe generally produced by Trichophyton rubrum. The aim of this study is to compare the traditional diagnostic techniques of superficial mycoses with a homemade and wide-spectrum fungal polymerase chain reaction (PCR) technique that amplifies a specific region of the 18S ribosomal RNA (rRNA) directly from samples of scales, nails and hair. A total of 626 clinical samples (obtained in the Basurto University Hospital, Bilbao, Spain) were analysed by traditional culture, microscopy and PCR. DNA extraction was carried out by using an extraction buffer and bovine serum, and amplification of samples and performance of the PCR were checked by conventional agarose gel electrophoresis with subsequent sequencing of amplified samples. A total of 211 samples (34%) resulted in positive diagnosis with at least one of the two applied methods: culture (21%) and PCR (22%). Despite the low percentage of identification achieved by the sequencing technique (40%), the value contributed by the amplification of the 18S region of the rRNA was considered important in the identification as it showed a high predictive values for both positive and negative diagnoses (90.9% and 94.6%, respectively). The proposed PCR method has been confirmed as a complementary, rapid, and effective method in the diagnosis of superficial mycoses. Additionally, it reduces the time to obtain satisfactory results from 4 weeks to 7 h.
Assuntos
Arthrodermataceae/isolamento & purificação , Dermatomicoses/diagnóstico , Cabelo/microbiologia , Técnicas de Diagnóstico Molecular/métodos , Unhas/microbiologia , Reação em Cadeia da Polimerase/métodos , Pele/microbiologia , Adulto , Idoso , Arthrodermataceae/classificação , Arthrodermataceae/genética , DNA Fúngico/genética , DNA Ribossômico/genética , Feminino , Humanos , Masculino , Técnicas Microbiológicas/métodos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Espanha , VoluntáriosRESUMO
PURPOSE: Fructooligosaccharides (FOS) are used as functional foods due to their prebiotic effects. Intestinal anti-inflammatory activity has been established in most, but not all, studies in animal models of colitis, using mainly chemically induced inflammation. Our goal was to test the effect of FOS (degree of polymerization 2-8) in the chronic, lymphocyte-driven CD4+ CD62L+ T cell transfer model of colitis. METHODS: Colitis was induced by transfer of CD4+ CD62L+ T cells to C57BL/6J Rag1(-/-) mice. FOS (75 mg day(-1)) was administered by gavage as a post-treatment. Three groups were established: non-colitic (NC), colitic control (C, CD4+ CD62L+ transferred mice treated with vehicle) and colitic+FOS (C+FOS, similar but treated with FOS). Mice were killed after 13 days. RESULTS: Treatment of mice with FOS ameliorated colitis, as evidenced by an increase in body weight, a lesser myeloperoxidase and alkaline phosphatase activities, a lower secretion of proinflammatory cytokines by mesenteric lymph node cells ex vivo (IFN-γ, IL-17, and TNF-α), and a higher colonic expression of occludin (C+FOS vs. C, p < 0.05). Increased relative abundance of lactic acid bacteria was observed in FOS-treated mice (p < 0.05). CONCLUSIONS: FOS exert intestinal anti-inflammatory activity in T lymphocyte-dependent colitis, suggesting it may be useful in the management of inflammatory bowel disease in appropriate conditions.
Assuntos
Anti-Inflamatórios/farmacologia , Colite/tratamento farmacológico , Intestinos/efeitos dos fármacos , Oligossacarídeos/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Linfócitos T CD4-Positivos/metabolismo , Calgranulina A/genética , Calgranulina A/metabolismo , Claudina-4/genética , Claudina-4/metabolismo , Claudina-5/genética , Claudina-5/metabolismo , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Feminino , Microbioma Gastrointestinal , Regulação da Expressão Gênica , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-17/genética , Interleucina-17/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/microbiologia , Selectina L/metabolismo , Lactobacillus , Camundongos , Camundongos Endogâmicos C57BL , Ocludina/genética , Ocludina/metabolismo , Peroxidase/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Heart failure is a health problem associated with disability and mortality. Physicians may stratify the risk of adult patients with heart failure using a cardiopulmonary exercise testing. Until now, in childhood this evaluation has been poorly used. The purpose of this study is to compare the peak oxygen uptake and minute ventilation/carbon dioxide production slope among children with heart failure versus children without heart disease (control). METHODS: Thirty-eight children with heart failure were compared with 194 children without heart disease. All of them performed cardiopulmonary exercise testing using a symptom-limited ramp protocol. Differences between groups were compared using Chi-squared test, Student's t test, or ANOVA. Any value of p < 0.05 was considered significant. RESULTS: Children with heart failure were older, taller, and with a higher prevalence of male gender. This group had also a lower peak oxygen uptake (27 ± 10 ml O2/kg/min) compared to the control group (37 ± 10 ml O2/kg/min); p < 0.001. The minute ventilation/carbon dioxide production was higher in the heart failure group (31 ± 4) than in controls (28 ± 6); p < 0.001. CONCLUSION: Children with heart failure showed lower peak oxygen uptake and higher minute ventilation/carbon dioxide production slope than the control group.
Assuntos
Dióxido de Carbono/metabolismo , Insuficiência Cardíaca/metabolismo , Consumo de Oxigênio , Respiração , Adolescente , Estatura , Estudos de Casos e Controles , Distribuição de Qui-Quadrado , Criança , Teste de Esforço , Feminino , Insuficiência Cardíaca/fisiopatologia , Humanos , Masculino , Prognóstico , Fatores SexuaisRESUMO
Flavonoids are polyphenolic compounds that are widespread in nature, and consumed as part of the human diet in significant amounts. The aim of the present study was to test the intestinal anti-inflammatory activity of apigenin K, a soluble form of apigenin, in two models of rat colitis, namely the trinitrobenzenesulfonic acid (TNBS) model and the dextran sulphate sodium (DSS) model. Apigenin K (1, 3 and 10 mg/kg; by the oral route; n 4-6 per group) was administered as a pre-treatment to rats with TNBS and DSS colitis, and colonic status was checked by macroscopic and biochemical examination. Apigenin K pre-treatment resulted in the amelioration of morphological signs and biochemical markers in the TNBS model. The results demonstrated a reduction in the inflamed area, as well as lower values of score and colonic weight:length ratio compared with the TNBS group. Myeloperoxidase (MPO) activity was reduced by 30 % (P< 0·05). Moreover, apigenin K pre-treatment ameliorated morphological signs and biochemical markers in the DSS model. Thus, macroscopic damage was significantly reduced and the colonic weight:length ratio was lowered by approximately 10 %, while colonic MPO and alkaline phosphatase activities were decreased by 35 and 21 %, respectively (P< 0·05). Apigenin K pre-treatment also tended to normalise the expression of a number of colonic inflammatory markers (e.g. TNF-α, transforming growth factor-ß, IL-6, intercellular adhesion molecule 1 or chemokine (C-C motif) ligand 2). In conclusion, apigenin K is found to have anti-inflammatory effects in two preclinical models of inflammatory bowel disease.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Apigenina/uso terapêutico , Colite/dietoterapia , Suplementos Nutricionais , Modelos Animais de Doenças , Fármacos Gastrointestinais/uso terapêutico , Doenças Inflamatórias Intestinais/dietoterapia , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/química , Apigenina/administração & dosagem , Apigenina/química , Biomarcadores/metabolismo , Colite/imunologia , Colite/metabolismo , Colite/patologia , Colo/imunologia , Colo/metabolismo , Colo/patologia , Sulfato de Dextrana , Feminino , Fármacos Gastrointestinais/administração & dosagem , Fármacos Gastrointestinais/química , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Tamanho do Órgão , Projetos Piloto , Distribuição Aleatória , Ratos Wistar , Solubilidade , Ácido TrinitrobenzenossulfônicoRESUMO
The universal screening for osteoporosis by bone mineral density (BMD) is not feasible because of its unfavorable cost-benefit due to its low sensitivity. The aim of the present study was to estimate the population and economic impact of the diagnostic criteria of the National Osteoporosis Guideline Group (NOGG) and the National Osteoporosis Foundation (NOF) and assess the appropriateness of the BMD tests performed in routine clinical practice. A cross-sectional study was conducted in individuals referred for BMD testing who were not receiving antiresorptive therapy. The absolute risk of major and hip fracture was calculated using the British formula of the Fracture Risk Assessment Tool. NOGG and NOF guidelines diagnostic thresholds interventions were used. A total of 640 individuals were included, of which 95% were women, with a median age of 59.4 years (interquartile range = 14). When applying the NOGG criteria, BMD testing was recommended in 32.3% of the individuals, whereas this percentage increased to 75.6% with the NOF guidelines (p < 0.05). Regarding the appropriateness of the BMD tests performed, 31.9% were deemed appropriate according to both the NOGG and NOF guidelines, whereas 23.9% were considered inappropriate. In conclusion, the application of the NOGG and NOF guidelines led to a decrease in BMD indications, reducing costs and improving efficiency in the diagnostic management of osteoporosis, although variability exists between the guidelines.
Assuntos
Densidade Óssea , Osteoporose/diagnóstico , Guias de Prática Clínica como Assunto , Idoso , Análise Custo-Benefício , Estudos Transversais , Feminino , Humanos , MasculinoRESUMO
Rutin, one of the most abundant flavonoids in nature, has been shown to exert intestinal antiinflammatory effects in experimental models of colitis. Our aim was to study the antiinflamatory effect of rutin in the CD4+ CD62L+ T cell transfer model of colitis, one of the closest to the human disease. Colitis was induced by transfer of CD4+ CD62L+ T cells to Rag1(-/-) mice. Rutin was administered by gavage as a postreatment. Treatment with rutin improved colitis at the dose of 57mg/kg/day, while no effect was noted with 28.5mg/kg/day. Therapeutic benefit was evidenced by a reduced disease activity index, weight loss and damage score, plus a 36% lower colonic myeloperoxidase and a 54% lower alkaline phosphatase activity. In addition, a decreased secretion of proinflammatory cytokines (IFNγ and TNFα) by mesenteric lymph node cells was observed ex vivo. The colonic expression of proinflammatory genes, including IFNγ, TNFα, CXCL1, S100A8 and IL-1ß, was significantly reduced by more than 80% with rutin as assessed by RT-qPCR. Flavonoid treated mice exhibited decreased activation of splenic CD4+ cells (STAT4 phosphorylation and IFNγ expression) and reduced plasma cytokine levels. This effect was also apparent in mucosal lymphocytes based on reduced STAT4 phosphorylation. The protective effect was comparable to that of 3mg/kg/day budesonide. Rutin had no effect on splenocytes or murine T cells in vitro, while its aglycone, quercetin, exhibited a concentration dependent inhibition of proinflammatory cytokines, including IFNγ. Rutin but not quercetin showed vectorial basolateral to apical transport in IEC18 cells, associated with reduced biotransformation. We conclude that rutin exerts intestinal antiinflammatory activity in chronic, T lymphocyte dependent colitis via quercetin release and actions involving mucosal and lymph node T cells. Our results suggest that rutin may be useful in the management of inflammatory bowel disease in appropriate dosage conditions.
Assuntos
Anti-Inflamatórios/uso terapêutico , Colite/tratamento farmacológico , Rutina/uso terapêutico , Fosfatase Alcalina/metabolismo , Animais , Anti-Inflamatórios/farmacologia , Linhagem Celular , Células Cultivadas , Colite/sangue , Colite/metabolismo , Colite/patologia , Citocinas/sangue , Citocinas/genética , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Proteínas de Homeodomínio/genética , Intestino Grosso/efeitos dos fármacos , Intestino Grosso/patologia , Linfonodos/citologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , NF-kappa B/metabolismo , Peroxidase/metabolismo , Quercetina/farmacologia , RNA Mensageiro/metabolismo , Ratos Wistar , Rutina/farmacologia , Fator de Transcrição STAT4/metabolismo , Baço/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismoRESUMO
Milk κ-casein-derived bovine glycomacropeptide (GMP) exerts immunomodulatory effects. It exhibits intestinal anti-inflammatory activity in chemically induced models of colitis. However, to validate its clinical usefulness as a nutraceutical, it is important to assess its effects in a model with a closer pathophysiological connection with human inflammatory bowel disease. Therefore, in the present study, we used the lymphocyte-transfer model of colitis in mice and compared the effects of GMP in this model with those obtained in the dextran sulphate sodium (DSS) model. GMP (15 mg/d) resulted in higher body-weight gain and a reduction of the colonic damage score and myeloperoxidase (MPO) activity in Rag1(-/-) mice with colitis induced by the transfer of naïve T cells. The colonic and ileal weight:length ratio was decreased by approximately 25%, albeit non-significantly. GMP treatment reduced the percentage of CD4⺠interferon (IFN)-γ⺠cells in mesenteric lymph nodes (MLN). The basal production of IL-6 by MLN obtained from the GMP-treated mice ex vivo was augmented. However, concanavalin A-evoked production was similar. The colonic expression of regenerating islet-derived protein 3γ, S100A8, chemokine (C-X-C motif) ligand 1 and IL-1ß was unaffected by GMP, while that of TNF-α and especially IFN-γ was paradoxically increased. In the DSS model, GMP also reduced the activity of colonic MPO, but it failed to alter weight gain or intestinal weight:length ratio. GMP augmented the production of IL-10 by MLN cells and was neutral towards other cytokines, except exhibiting a trend towards increasing the production of IL-6. The lower effect was attributed to the lack of the effect of GMP on epithelial cells. In conclusion, GMP exerts intestinal anti-inflammatory effects in lymphocyte-driven colitis.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Caseínas/uso terapêutico , Suplementos Nutricionais , Modelos Animais de Doenças , Fármacos Gastrointestinais/uso terapêutico , Doenças Inflamatórias Intestinais/prevenção & controle , Mucosa Intestinal/imunologia , Fragmentos de Peptídeos/uso terapêutico , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Bovinos , Colo/imunologia , Colo/metabolismo , Colo/patologia , Feminino , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Íleo/imunologia , Íleo/metabolismo , Íleo/patologia , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/metabolismo , Doenças Inflamatórias Intestinais/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Linfadenite Mesentérica/etiologia , Linfadenite Mesentérica/prevenção & controle , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infiltração de Neutrófilos , Peroxidase/sangue , Peroxidase/metabolismo , Distribuição Aleatória , Aumento de PesoRESUMO
Uncontrolled bleeding during surgery is associated with high mortality and prolonged hospital stay, necessitating the use of hemostatic agents. Fibrin sealant patches offer an efficient solution to achieve hemostasis and improve patient outcomes in liver resection surgery. We have previously demonstrated the efficacy of a nanostructured fibrin-agarose hydrogel (NFAH). However, for the widespread distribution and commercialization of the product, it is necessary to develop an optimal preservation method that allows for prolonged stability and facilitates storage and distribution. We investigated cryopreservation as a potential method for preserving NFAH using trehalose. Structural changes in cryopreserved NFAH (Cryo-NFAH) were investigated and comparative in vitro and in vivo efficacy and safety studies were performed with freshly prepared NFAH. We also examined the long-term safety of Cryo-NFAH versus TachoSil in a rat partial hepatectomy model, including time to hemostasis, intra-abdominal adhesion, hepatic hematoma, inflammatory factors, histopathological variables, temperature and body weight, hemocompatibility and cytotoxicity. Structural analyses demonstrated that Cryo-NFAH retained most of its macro- and microscopic properties after cryopreservation. Likewise, hemostatic efficacy assays showed no significant differences with fresh NFAH. Safety evaluations indicated that Cryo-NFAH had a similar overall profile to TachoSil up to 40 days post-surgery in rats. In addition, Cryo-NFAH demonstrated superior hemostatic efficacy compared with TachoSil while also demonstrating lower levels of erythrolysis and cytotoxicity than both TachoSil and other commercially available hemostatic agents. These results indicate that Cryo-NFAH is highly effective hemostatic patch with a favorable safety and tolerability profile, supporting its potential for clinical use.
Assuntos
Criopreservação , Hemostáticos , Hidrogéis , Nanoestruturas , Sefarose , Animais , Hidrogéis/química , Hemostáticos/farmacologia , Hemostáticos/química , Ratos , Sefarose/química , Criopreservação/métodos , Nanoestruturas/química , Fibrina/química , Masculino , Hepatectomia/métodos , Humanos , Hemostasia/efeitos dos fármacos , Ratos Sprague-DawleyRESUMO
Diarrhoea is a hallmark of intestinal inflammation. The mechanisms operating in acute inflammation of the intestine are well characterized and are related to regulatory changes induced by inflammatory mediators such as prostaglandins, cytokines or reactive oxygen species, along with leakage due to epithelial injury and changes in permeability. In chronic colitis, however, the mechanisms are less well known, but it is generally accepted that both secretory and absorptive processes are inhibited. These disturbances in ionic transport may be viewed as an adaptation to protracted inflammation of the intestine, since prolonged intense secretion may be physiologically unacceptable in the long term. Mechanistically, the changes in transport may be due to adjustments in the regulation of the different processes involved, to broader epithelial alterations or frank damage, or to modulation of the transportome in terms of expression. In the present review, we offer a summary of the existing evidence on the status of the transportome in chronic intestinal inflammation.
Assuntos
Colite/metabolismo , Enterócitos/metabolismo , Absorção Intestinal , Animais , Colite/patologia , Colite/fisiopatologia , Enterócitos/patologia , Expressão Gênica , Humanos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismoRESUMO
Objective: Although physical training has been associated with an increase in survival, its role in reducing Exercise-induced arrhythmias (EIA) in patients with heart disease has not yet been dilucidated. We aim to compare the effect of physical training on the occurrence of EIA in patients with heart disease. Methodology: We evaluated a retroprospective and self-controlled cohort of patients older than 18 years with heart disease who entered the cardiac rehabilitation program of the National Institute of Cardiology Ignacio Chávez in México, during January 2015 to December 2016. In all patients, cardiovascular risk was stratified, including a cardiopulmonary exercise test, which was also performed at the end of the program. The occurrence of arrhythmias was evaluated before and after the training program. Results: 160 patients were analyzed, of which 126 (79%) were male. The proportion of patients who developed EIA in the first exercise testing was 56% and in the second one was 48%. In the first group 35% of patients did not developed EIA in the second test. Of the 71 patients who did not present EIA at the beginning of the program, twenty (28%) presented arrhythmias at second test. In relation to the effect of the training program on the occurrence of arrhythmias, we could observe that it had a protective effect, with a RR = 0.49 (95% CI: 0.35-0.67, p <0.001). Conclusion: In this study, physical training was associated with a decreased risk of developing EIA.
RESUMO
Milk κ-casein-derived bovine glycomacropeptide (GMP) has immunomodulatory and bacterial toxin-binding effects, and it has been shown to exert intestinal antiinflammatory activity in the trinitrobenzenesulfonic acid-induced model of colitis. However, its mechanism of action is not well characterized, and it is not known whether GMP is effective in other experimental models. The intestinal antiinflammatory activity of GMP was assessed in the dextran sulfate sodium (DSS)-induced model of rat colitis. DSS was applied at a starting concentration of 5% (wt:v) in drinking water and adjusted when the disease activity index (DAI) increased substantially for 10 d. There were 3 experimental groups: control (no inflammation), DSS, and GMP (GMP-treated rats with DSS-induced colitis). GMP pretreatment (500 mg · kg(-1) · d(-1), starting 2 d before DSS treatment) reduced the DAI by 60% and lowered the colonic damage score by 44% (P < 0.05). GMP fully normalized the colonic expression of interleukin (IL) 1ß, IL17, IL23, IL6, transforming growth factor ß, IL10, and Foxp3 as assessed by quantitative RT-PCR. The production of interferon-γ by mesenteric lymph node cells ex vivo was also normalized by GMP treatment. In contrast, GMP did not change colonic thickening, myeloperoxidase, cyclooxygenase 2, or alkaline phosphatase. Histology analysis showed better preservation of the epithelium and attenuated infiltration and submucosal thickening in rats treated with GMP. We conclude that GMP exerts intestinal antiinflammatory activity in this model, which may be primarily related to actions on Th1 and Th17 lymphocytes and perhaps macrophages.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Caseínas/farmacologia , Caseínas/uso terapêutico , Colite/prevenção & controle , Glicopeptídeos/farmacologia , Glicopeptídeos/uso terapêutico , Animais , Células Cultivadas , Colite/induzido quimicamente , Colite/metabolismo , Colo/efeitos dos fármacos , Colo/metabolismo , Colo/patologia , Ciclo-Oxigenase 2/metabolismo , Feminino , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/metabolismo , Doenças Inflamatórias Intestinais/tratamento farmacológico , Doenças Inflamatórias Intestinais/fisiopatologia , Linfonodos/citologia , Linfonodos/efeitos dos fármacos , Linfonodos/metabolismo , Infiltração de Neutrófilos/efeitos dos fármacos , RNA Mensageiro/metabolismo , Distribuição Aleatória , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Índice de Gravidade de DoençaRESUMO
It is becoming standard practice to measure a housekeeping gene, typically actin, in Western blots, as it is the rule in RNA blots. We have applied reversible Ponceau staining to check equal loading of gels and measured actin in parallel under different conditions. Our results show that densitometric analysis is comparable with both techniques. Therefore, routine quantitation of Ponceau staining before antibody probing is validated as an alternative to actin blotting.
Assuntos
Actinas/análise , Western Blotting/métodos , Coloração e Rotulagem/métodos , Animais , Linhagem Celular , Colo/química , Humanos , Rim/química , Fígado/química , Camundongos , RatosRESUMO
The combined effect of farm management practices, transport time, and ageing time on the electrophoretic changes of sarcoplasmic (SPP) and myofibrillar (MFP) protein fractions of goat kids was studied. A total of 64 suckling goat kids were withdrawn from two farms with "high" (GW) and "low" (DW) welfare-friendly management practices, and they were transported for 2 or 6 h immediately before slaughtering. Longissimus lumborum samples were obtained at 3, 8, and 21 days post-mortem, and muscle proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis SDS-PAGE. Both protein extracts displayed significant changes attributable to meat maturation. Managing conditions of kids in DW farms increased the post-mortem susceptibility of muscle proteins. Some MFP of Longissimus lumborum muscle, such as troponin T, as well as 26-30 and 35-37 kDa fractions were influenced significantly by deficient on-farm management, and therefore, these protein fragments might be considered as indicators of low-welfare on-farm management in goat kids.
RESUMO
Dog overpopulation is considered a human health risk; they are the terrestrial vector of rabies and reservoirs for other human diseases. Surgical neutering and intratesticular injections have been used in male dogs. Physiological and morphological alterations in reproductive organs can be induced by phytoestrogens. Our goal was to evaluate the effect of oral coumestrol on dog ejaculates and testis histology. Two groups of 5 healthy adult dogs were used. One coumestrolcontaining biscuit was given once a week for a 4 week period to the experimental group. Ejaculates were obtained and evaluated. After treatment, testis were obtained and processed for histology. Compared to controls, treated dogs have reduced tubules (462 +/- 1.4 vs 336 +/- 2 micron(2)), spermatogenic epithelium (49.1 +/- 0.01 vs 13.3 +/- 0.01 micron(2)), and lumen opening (891 +/- 1.4 vs 530 +/- 26.9 micron). Ejaculates from treated animals have increased numbers of abnormal spermatozoa and reduced sperm concentration.
Assuntos
Anticoncepcionais Masculinos/farmacologia , Cumestrol/farmacologia , Fitoestrógenos/farmacologia , Testículo/efeitos dos fármacos , Animais , Anticoncepcionais Masculinos/administração & dosagem , Cumestrol/administração & dosagem , Cães , Masculino , Fitoestrógenos/administração & dosagem , Contagem de Espermatozoides/veterináriaRESUMO
BACKGROUND: Trinitrobenzenesulphonic acid (TNBS) induced rat colitis is one of the most widely used models of inflammatory bowel disease (IBD), a condition whose aetiology and pathophysiology are incompletely understood. We have characterized this model at the genomic level using a longitudinal approach. Six control rats were compared with colitic animals at 2, 5, 7 and 14 days after TNBS administration (n = 3). The Affymetrix Rat Expression Array 230 2.0 system was used. RESULTS: TNBS-induced colitis had a profound impact on the gene expression profile, which was maximal 5 and 7 days post-induction. Most genes were affected at more than one time point. They were related to a number of biological functions, not only inflammation/immunity but also transport, metabolism, signal transduction, tissue remodeling and angiogenesis. Gene changes generally correlated with the severity of colitis. The results were successfully validated in a subset of genes by real-time PCR. CONCLUSION: The TNBS model of rat colitis has been described in detail at the transcriptome level. The changes observed correlate with pathophysiological disturbances such as tissue remodelling and alterations in ion transport, which are characteristic of both this model and IBD.
Assuntos
Colite/induzido quimicamente , Colo/efeitos dos fármacos , Perfilação da Expressão Gênica/métodos , Ácido Trinitrobenzenossulfônico/toxicidade , Animais , Análise por Conglomerados , Colite/genética , Colo/fisiopatologia , Modelos Animais de Doenças , Feminino , Genômica , Doenças Inflamatórias Intestinais/induzido quimicamente , Doenças Inflamatórias Intestinais/genética , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Ratos Wistar , Fatores de TempoRESUMO
INTRODUCTION: Estrogens are well recognized as important hormones in male reproduction and act as ligands to alpha and beta estrogen receptors. Both estrogen receptors could interact with estrogen-mimicking compounds such as the fluorescent phytoestrogen coumestrol, which acts both in an agonist or antagonist fashion. OBJECTIVE: To investigate the presence of Coumestrol-Estrogen Receptor complexes by fluorescence in testis and epididymis, its effect in the ER expression by immunostain in the same tissues and the effect of this binding in the testis histological characteristics. DESIGN: Adult healthy and sexually active dogs were assigned to either the experimental or control group .Coumestrol impregnated dog biscuits were given to each animal from the experimental group once a week for a 4 week period. The control group received a biscuit with no Coumestrol, also once a week and for the same period. Testis morphology, ER immunodetection, and coumestrol-receptor binding were evaluated. SETTING: The experiment was done in the facilities of the Mexico City canine shelter. Animals were caged individually with food and water ad libitum and having at least two daily hours for exercise. RESULTS: Morphological alterations in testis after oral administration of coumestrol were detected. The main alterations include decreased germinal epithelium in tubule, and the loss of a continuous proliferation and differentiation gamete layer. Fluorescence signals in testis interstitial Leydig cells and epididymus indicating ER-coumestrol complexes were detected at the same points to those Immunohystochemically detected ER. CONCLUSIONS: Coumestrol administration induces testis alterations and coumestrol-ER complexes can be co-localized by binding-enhanced fluorescence and immunoprecipitation.
Assuntos
Cumestrol/farmacologia , Epididimo/efeitos dos fármacos , Fitoestrógenos/farmacologia , Receptores de Estrogênio/metabolismo , Testículo/efeitos dos fármacos , Administração Oral , Animais , Cumestrol/administração & dosagem , Cães , Epididimo/metabolismo , Masculino , Fitoestrógenos/administração & dosagem , Distribuição Aleatória , Receptores de Estrogênio/efeitos dos fármacos , Túbulos Seminíferos/efeitos dos fármacos , Testículo/metabolismoRESUMO
BACKGROUND AND PURPOSE: Calprotectin is a heterodimer composed of two myeloid-related proteins, S100A8 and S100A9, that is abundant in neutrophils and monocytes/macrophages. Faecal levels of calprotectin are used routinely to monitor inflammatory bowel disease activity. EXPERIMENTAL APPROACH: We aimed to assess the role of calprotectin in intestinal inflammation, using the dextran sulfate sodium model of colitis in mice. Calprotectin was administered (50 or 100 µg·day-1 ) by the intrarectal or by i.p. injection (50 µg·day-1 only). The condition of the mice was characterized by morphological and biochemical methods. KEY RESULTS: Intrarectal calprotectin protected significantly against colitis, as shown by lower levels of macroscopic and microscopic damage, colonic myeloperoxidase activity and decreased expression of TNFα and toll-like receptor 4. IL-17 production by spleen and mesenteric lymph node cells was reduced. Calprotectin had no effect on body weight loss or colonic thickening. There were no effects of calprotectin after i.p. injection. Calprotectin had virtually no effects in control, non-colitic mice. Calprotectin had almost no effect on the colonic microbiota but enhanced barrier function. Treatment of rat IEC18 intestinal epithelial cells in vitro with calprotectin induced output of the chemokines CXL1 and CCL2, involving the receptor for advanced glycation end products- and NFκB. CONCLUSION AND IMPLICATIONS: Calprotectin exerted protective effects in experimental colitis when given by the intrarectal route, by actions that appear to involve effects on the epithelium.
Assuntos
Colite/prevenção & controle , Inflamação/prevenção & controle , Complexo Antígeno L1 Leucocitário/farmacologia , Administração Retal , Animais , Células Cultivadas , Colite/induzido quimicamente , Colite/tratamento farmacológico , Colite/patologia , Sulfato de Dextrana/antagonistas & inibidores , Modelos Animais de Doenças , Feminino , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/patologia , Complexo Antígeno L1 Leucocitário/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BLRESUMO
OBJECTIVE: Exercise-induced premature ventricular complexes (EiPVCs) are often considered as benign arrhythmias, although they are associated with a high risk of all-cause death in the general healthy population. However, an intermediate pathophysiological process remains unclear, particularly in patients with known cardiovascular disease. The aim of this study was to find an association between EiPVCs, the occurrence of life-threatening ventricular arrhythmias (LACO), and all-cause mortality in patients with cardiovascular disease. METHODS: This was an observational study of a cohort of patients with coronary artery disease (CAD) or idiopathic cardiomyopathy (ICM). Stress testing was performed as a part of the routine cardiovascular evaluation. The occurrence of EiPVCs was evaluated during exercise testing (ET). At follow-up, long-term occurrence of LACO was evaluated. A bivariate and multivariate analysis was performed. RESULTS: Out of the total of 1442 patients analysed, 700 (49%) had EiPVCs. During 14 years of following-up after ET, 106 LACO outcomes were observed. Long-term all-cause mortality was 4% (n=61). A bivariate analysis showed that patients with EiPVCs had an increased risk for LACO (RR=2.81, 95% CI; 1.9-4.3, P<.001), and for mortality (RR=2.1, CI95% 1.2-3.4, P<.01). Occurrence of LACO was also associated with a higher mortality risk (RR=5.7, 95% CI; 3.4-9.4, P<.001). After a post hoc analysis, LACO remained as a highly predictive variable for mortality. CONCLUSION: Patients with EiPVCs have a high risk of LACO and all-cause mortality. The presence of LACO could be an intermediate stage between EiPVCs and mortality in subjects with cardiovascular disease.