RESUMO
The aim of this study was to develop a simple method to purify Strongyloides eggs from rat faeces using a sucrose gradient centrifugal-flotation technique. This procedure is simple, rapid and possesses a high efficiency in recovering Strongyloides eggs without faecal detritus in less than one hour, thus eliminating the use of complex apparatus and different chemical substances. The possibility of working with pure and live Strongyloides eggs opens up a wide range of future studies on the biology of this parasite. This study constitutes the first report in the scientific literature on purifying Strongyloides eggs using a sucrose density gradient.
Assuntos
Fezes/parasitologia , Contagem de Ovos de Parasitas/métodos , Ratos/parasitologia , Strongyloides/isolamento & purificação , AnimaisRESUMO
The present study suggests that insulin resistance has no association with bone quantity, but quality. INTRODUCTION: The literature has contradictory results concerning the influence of insulin resistance on bone. The present study sought to evaluate the association of insulin resistance and adipose tissue with either bone mineral density or the trabecular bone score. METHODS: The study included 56 individuals (36 women and 20 men): age = 46.6 ± 14.2 years, weight = 67.8 ± 10.9 kg, height = 1.65 ± 0.10 m and BMI = 24.8 ± 3.9 kg/m2. The investigational protocol included biochemical determinations and bone assessment by dual X-ray absorptiometry for evaluation of bone mineral density and trabecular bone score. Magnetic resonance was employed to estimate visceral, subcutaneous and bone marrow adipose tissues, as well as intrahepatic lipids. RESULTS: The bone mineral density of the lumbar spine, femoral neck and total hip were not associated with insulin resistance-related parameters [visceral adipose tissue, intrahepatic lipids and homeostatic model assessment of insulin resistance (HOMA-IR)]. In contrast, there was a negative relationship between the trabecular bone score and all these components. The association between the trabecular bone score and HOMA-IR was reinforced after adjustment for age and BMI. Marrow adipose tissue was negatively associated with both bone mineral density and trabecular bone score. CONCLUSIONS: The present study shows that the trabecular bone score is negatively associated with marrow adipose tissue, insulin resistance, visceral adipose tissue and intrahepatic lipid measurements. Additionally, there was a negative relationship between saturated lipids in marrow adipose tissue and the trabecular bone score. These results encourage further studies to investigate the role of the trabecular bone score exam in the clinical evaluation of osteoporosis in conditions of insulin resistance.
Assuntos
Tecido Adiposo/diagnóstico por imagem , Densidade Óssea , Osso Esponjoso/diagnóstico por imagem , Resistência à Insulina , Absorciometria de Fóton , Adulto , Medula Óssea , Feminino , Humanos , Gordura Intra-Abdominal , Lipídeos/análise , Fígado/química , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-IdadeRESUMO
The mechanisms involved in kidney disturbances during development, induced by vitamin D3 deficiency in female rats, that persist into adulthood were evaluated in this study. Female offspring from mothers fed normal (control group, n=8) or vitamin D-deficient (Vit.D-, n=10) diets were used. Three-month-old rats had their systolic blood pressure (SBP) measured and their blood and urine sampled to quantify vitamin D3 (Vit.D3), creatinine, Na+, Ca+2 and angiotensin II (ANGII) levels. The kidneys were then removed for nitric oxide (NO) quantification and immunohistochemical studies. Vit.D- pups showed higher SBP and plasma ANGII levels in adulthood (P<0.05) as well as decreased urine osmolality associated with increases in urinary volume (P<0.05). Decreased expression of JG12 (renal cortex and glomeruli) and synaptopodin (glomeruli) as well as reduced renal NO was also observed (P<0.05). These findings showed that renal disturbances in development in pups from Vit.D- mothers observed in adulthood may be related to the development of angiogenesis, NO and ANGII alterations.
Assuntos
Nefropatias/etiologia , Rim/irrigação sanguínea , Deficiência de Vitamina D/complicações , Animais , Feminino , Rim/metabolismo , Rim/patologia , Nefropatias/metabolismo , Nefropatias/patologia , Fenômenos Fisiológicos da Nutrição Materna , RatosRESUMO
OBJECTIVES: Oral lichen planus is a chronic, T-cell-mediated, inflammatory disease that affects the oral cavity. The oral lichen planus pathogenesis is still unclear, however, the main evidence is that the mechanisms of activation of different T lymphocyte pathway induce apoptosis with an increase in Th1 and Th17 subtypes cells, triggered by the release of cytokines. This study analysed saliva proteomics to identify protein markers that might be involved in the pathogenesis and development of the disease. MATERIAL AND METHODS: Proteins differentially expressed by oral lichen planus and healthy controls were screened using mass spectrometry; the proteins found in oral lichen planus were subjected to bioinformatics analysis, including gene ontology and string networks analysis. The multiplex analysis validation allowed the correlation between the proteins identified and the involved cytokines in Th17 response. RESULTS: One hundred and eight proteins were identified in oral lichen planus, of which 17 proteins showed a high interaction between them and indicated an association with the disease. Expression of these proteins was correlated with the triggering of cytokines, more specifically the Th17 cells. CONCLUSION: Proteins, such as S100A8, S100A9, haptoglobin, can trigger cytokines and might be associated with a pathological function and antioxidant activities in oral lichen planus.
Assuntos
Líquen Plano Bucal/metabolismo , Proteínas/metabolismo , Saliva/metabolismo , Estudos de Casos e Controles , Biologia Computacional , Citocinas/metabolismo , Feminino , Ontologia Genética , Humanos , Masculino , Pessoa de Meia-Idade , Proteoma , Células Th17/metabolismoRESUMO
Over the years, with the instrumental analysis evolution, the relationships between the carried-out results with the data of theoretical analysis in silico and the Hammett's parameters have been reported. They have been very useful for chemical characterization of small organic molecules. Thus, this work aims at showing the feasibility and limitations for Hammett's and density functional theory applications in electrospray ionization-collision-induced dissociation (ESI-CID) fragmentation provision. For this, 13 dihydropyrimidinones para, meta, and orto monosubstituted were studied using ESI and CID in positive mode. As a result, it was observed that the main fragmentation includes the isocyanate and ethanol loses at low energy. Nevertheless, at higher energies, radical ions formed by McLafferty rearrangement were observed. The Hammett plots were correlated fragmentation profiles, showing good linearity for the [M + H]+ , which does not occur to radical ions and carbocation's. These tendencies had demonstrated that the stability of protonate and activation energy of secondary ions changes with the pKa. The density functional theory studies indicated that, both nitrogen atoms in the dihydropyrimidinone's prototypes are capable of being protonated. However, the activation energy of fragmentation products is not changed. Therefore, this work has shown information, which can be useful to understand tandem mass spectrometry in ESI-CID conditions for small organic molecules series. This is the first step for normalization of fragmentation pathway.
Assuntos
Di-Hidropiridinas/química , Simulação por Computador , Di-Hidropiridinas/síntese química , Estrutura Molecular , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em TandemRESUMO
Bone marrow adipose tissue has not been studied in patients with inactive inflammatory bowel disease. We found that these patients have preserved marrow adiposity even with low bone mass. Factors involved in bone loss in active disease may have long-lasting effects but do not seem to affect bone marrow adiposity. INTRODUCTION: Reduced bone mass is known to occur at varying prevalence in patients with inflammatory bowel diseases (IBD) because of inflammation, malnutrition, and steroid therapy. Osteoporosis may develop in these patients as the result of an imbalanced relationship between osteoblasts and adipocytes in bone marrow. This study aimed to evaluate for the first time bone mass and bone marrow adipose tissue (BMAT) in a particular subgroup of IBD patients characterized by long-term, steroid-free remission. METHODS: Patients with Crohn's disease (CD; N = 21) and ulcerative colitis (UC; N = 15) and controls (C; N = 65) underwent dual X-ray energy absorptiometry and nuclear magnetic resonance spectroscopy of the L3 lumbar vertebra for BMAT assessment. RESULTS: Both the CD and UC subgroups showed significantly higher proportions of patients than controls with Z-score ≤-2.0 at L1-L4 (C 1.54%; CD 19.05%; UC 20%; p = 0.02), but not at other sites. The proportions of CD patients with a T-score Ë-1.0 at the femoral neck (C 18.46%; CD 47.62%; p = 0.02) and total hip (C 16.92%; CD 42.86%; p = 0.03) were significantly higher than among controls. There were no statistically significant differences between IBD patients and controls regarding BMAT at L3 (C 28.62 ± 8.15%; CD 29.81 ± 6.90%; UC 27.35 ± 9.80%; p = 0.67). CONCLUSIONS: IBD patients in long-term, steroid-free remission may have a low bone mass in spite of preserved BMAT. These findings confirm the heterogeneity of bone disorders in IBD and may indicate that factors involved in bone loss in active disease may have long-lasting effects on these patients.
Assuntos
Tecido Adiposo/patologia , Medula Óssea/patologia , Doenças Inflamatórias Intestinais/complicações , Osteoporose/etiologia , Absorciometria de Fóton/métodos , Adulto , Densidade Óssea/fisiologia , Estudos de Casos e Controles , Colite Ulcerativa/complicações , Colite Ulcerativa/patologia , Colite Ulcerativa/fisiopatologia , Doença de Crohn/complicações , Doença de Crohn/patologia , Doença de Crohn/fisiopatologia , Feminino , Colo do Fêmur/fisiopatologia , Articulação do Quadril/fisiopatologia , Humanos , Doenças Inflamatórias Intestinais/patologia , Doenças Inflamatórias Intestinais/fisiopatologia , Vértebras Lombares/fisiopatologia , Imageamento por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Osteoporose/patologia , Osteoporose/fisiopatologia , Adulto JovemRESUMO
The effects of jump training on bone structure before and after ovariectomy-induced osteopenia in rats were investigated. Jumping exercise induced favorable changes in bone mineral density, bone mechanical properties, and bone formation/resorption markers. This exercise is effective to prevent bone loss after ovariectomy even when osteopenia is already established. INTRODUCTION: The present study investigated the effects of jump training on bone structure before and after ovariectomy-induced osteopenia in 80 10-week-old Wistar rats. METHODS: Forty rats (prevention program) were randomly allocated to one of four equal groups (n = 10): sham-operated sedentary (SHAM-SEDp), ovariectomized (OVX) sedentary (OVX-SEDp), sham-operated exercised (SHAM-EXp), and OVX exercised (OVX-EXp). SHAM-EXp and OVX-EXp animals began training 3 days after surgery. Another 40 rats (treatment program) were randomly allocated into another four groups (n = 10): sham-operated sedentary (SHAM-SEDt), OVX sedentary (OVX-SEDt), sham-operated exercised (SHAM-EXt), and OVX exercised (OVX-EXt). SHAM-EXt and OVX-EXt animals began training 60 days after surgery. The rats in the exercised groups jumped 20 times/day, 5 days/week, to a height of 40 cm for 12 weeks. At the end of the experimental period, serum osteocalcin, follicle-stimulating hormone (FSH) dosage, dual X-ray absorptiometry (DXA), histomorphometry, and biomechanical tests were analyzed. RESULTS: The OVX groups showed higher values of FSH and body weight (p < 0.05). DXA showed that jump training significantly increased bone mineral density of the femur and fifth lumbar vertebra (p < 0.05). The stiffness of the left femur and fifth lumbar vertebra in the exercised groups was greater than that of the sedentary groups (p < 0.05). Ovariectomy induced significant difference in bone volume (BV/TV, percent), trabecular separation (Tb.Sp, micrometer), and trabecular number (Tb.N, per millimeter) (p < 0.05) compared to sham operation. Jump training in the OVX group induced significant differences in BV/TV, Tb.Sp, and Tb.N and decreased osteoblast number per bone perimeter (p < 0.05) compared with OVX nontraining, in the prevention groups. Osteocalcin dosage showed higher values in the exercised groups (p < 0.05). CONCLUSIONS: Jumping exercise induced favorable changes in bone mineral density, bone mechanical properties, and bone formation/resorption markers. Jump training is effective to prevent bone loss after ovariectomy even when osteopenia is already established.
Assuntos
Densidade Óssea/fisiologia , Doenças Ósseas Metabólicas/reabilitação , Terapia por Exercício/métodos , Absorciometria de Fóton , Animais , Fenômenos Biomecânicos , Peso Corporal/fisiologia , Doenças Ósseas Metabólicas/fisiopatologia , Doenças Ósseas Metabólicas/prevenção & controle , Remodelação Óssea/fisiologia , Feminino , Fêmur/fisiopatologia , Hormônio Foliculoestimulante/sangue , Vértebras Lombares/fisiopatologia , Ovariectomia , Condicionamento Físico Animal/métodos , Distribuição Aleatória , Ratos WistarRESUMO
ããOBJECTIVE: To identify which sperm characteristics were able to predict more accurately the quality of curimba (Prochilodus lineatus) semen upon freezing using canonical correlation analysis. METHODS: Eleven fish breeders with initial mean weight of 705.21 ± 111 g were used. For cryopreservation, 200 µL of semen were taken from each animal and diluted in the cryoprotectant solution (10% dimethyl sulfoxide and 5% Beltsville Thawing Solution Minitub) in a 1:4 ratio and placed into 0.5-mL straws. Sperm characteristics (motility, sperm abnormalities, total antioxidant activity and lipid peroxidation) were evaluated. A randomized block design with duplicate samples per treatment (fresh and frozen semen) was used. The block factor was the animals, and the experimental unit the ejaculates. Canonical correlation was used to evaluate the association between sperm characteristics of fresh semen and thawed semen. RESULT: There was a significant association (P = 0.10) among the variables measured in fresh semen with the variables measured in thawed semen, and 78.6% of the difference observed in the thawed semen can be attributed to variation of variables measured in fresh semen. Sperm motility, motility duration and antioxidant activity of the thawed semen showed an inverse relationship with those of the fresh semen; whereas the minor sperm abnormalities, major sperm abnormalities and lipid peroxidation showed a direct relationship with those of the fresh semen. Only the rate and motility duration of the thawed semen presented high correlation (-0.63 and -0.73, respectively) with the canonical variable represented by the sperm characteristics of fresh semen. CONCLUSIONS: The rate and motility duration of fresh semen may be used to predict the quality of the thawed sperm in Prochilodus lineatus.
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Caraciformes/fisiologia , Criopreservação/métodos , Preservação do Sêmen , Sêmen/fisiologia , Espermatozoides/fisiologia , Estatística como Assunto , Animais , Sobrevivência Celular/efeitos dos fármacos , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Masculino , Sêmen/efeitos dos fármacos , Análise do Sêmen , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacosRESUMO
Degenerative changes of the endometrium are directly related to age and fertility in mares. Chronic degenerative endometritis (CDE) is correlated with uterine fluid retention and reduced ability to clear uterine inflammation. Recent research in the areas of equine surgery and sports medicine has shown that platelet-rich plasma (PRP) treatment acts as an immunomodulator of the inflammatory response. Therefore, the aim of this study was to determine if the uterine infusion of PRP could modulate the local inflammatory response and modify the intrauterine NO concentrations after artificial insemination (AI) in both normal mares and those with CDE. Thirteen mares with endometrium classified as grade III on the histology (mares with CDE) and eight mares with endometrial histological classification I or II-a normal mares were selected to investigate the effect of PRP therapy. The mares were inseminated with fresh semen in two consecutive cycles in a crossover study design. Thereby, each mare served as its own control and the treatment was performed with intrauterine PRP infusion four hours after AI. The percentage of neutrophils in uterine cytology (CIT, %), uterine fluid accumulation observed on ultrasonography (FLU, mm) and nitric oxide concentration of uterine fluid (NO, µM) were analyzed before and 24 hours after AI. The results reported that mares with CDE (CIT, 68.3 ± 3.27, FLU, 10.7 ± 1.61) have a higher (P < 0.05) intrauterine inflammatory response after AI than normal mares (CIT, 24.4 ± 3.56, FLU, 0), but NO concentrations did not differ (P > 0.05) between categories of mares. In treated cycles with PRP, the intrauterine inflammatory response decrease (P < 0.05) in CDE mares (CDE: CIT, 31.4 ± 6.48, FLU, 5.5 ± 1.28; normal mares: CIT, 13.5 ± 4.31, FLU, 0) when compared with nontreated cycle (CDE: CIT, 68.3 ± 3.27, FLU, 10.7 ± 1.61; NM: CIT, 24.4 ± 3.56, FLU, 0), but did not modify NO concentrations in uterine fluid. Thus, we can conclude that PRP was effective in modulating the exacerbated uterine inflammatory response to semen in mares with CDE but did not reduce NO concentrations in intrauterine fluid.
Assuntos
Endometrite/veterinária , Doenças dos Cavalos/terapia , Inflamação/metabolismo , Plasma Rico em Plaquetas , Animais , Estudos de Casos e Controles , Endometrite/terapia , Feminino , CavalosRESUMO
INTRODUCTION: In the intensive care unit (ICU), mortality is considered higher among renal transplant recipients than among nontransplantation patients. However, data regarding severe complications after kidney transplantation are scarce. MATERIALS AND METHODS: In this study, we evaluated all consecutive renal transplant recipients admitted to our ICU between July 2012 and July 2013 (n = 70), comparing their outcomes with those of a control group of nontransplantation patients admitted during the same period (n = 153). Among the transplant recipients, we compared survivors and nonsurvivors to identify predictors of ICU mortality. RESULTS: The mean age of the transplant recipients was 52 ± 13 years. Of the 70 transplant recipients, 18 (25%) required mechanical ventilation, 28 (40%) required inotropic support, and 27 (39%) required hemodialysis, all of which are factors that worsen the prognosis significantly. Twenty-two (31%) of the transplant recipients died in the ICU and 17 (24%) died within 30 days after ICU discharge, rates similar to those observed for the control group. CONCLUSIONS: We observed similar mortality between recipient and control groups, albeit the mortality was higher in the clinical group. In the multivariate model, the need for mechanical ventilation and the need for hemodialysis were independently associated with mortality.