ABSTRACT
O objetivo deste trabalho foi avaliar a recuperação de espermatozoides epididimários de cães castrados, utilizando as técnicas de fluxo retrógrado (FR) e flutuação (FL) em diluidor Tris-gema, antes e após a criopreservação. Foram coletados 30 complexos testículo-epididímos (CTE), sendo 15 para FR e 15 para FL, e, logo após a recuperação dos espermatozoides, foram analisadas as alterações morfológicas nessas células espermáticas. Após a adição do diluidor, foram avaliados os parâmetros de motilidade total (MOT) e vigor (V) espermáticos. O sêmen pós-criopreservado foi submetido ao teste de termorresistência nos tempos T0, T30, T60 e T90 minutos, além da avaliação das membranas plasmática e acrossomal por sondas fluorescentes. Não houve diferença estatística entre as técnicas quanto à MOT e ao vigor no sêmen diluído (FR-MOT: 82,3% e V: 3,4; FL-MOT: 79,6% e V: 3,2) e pós-criopreservado (FR-MOT: 34% e V: 2,8; FL-MOT: 30% e V: 2,7). A partir do T30, houve diferença significativa quanto à MOT e ao vigor nas técnicas utilizadas, e o tempo também prejudicou o acrossoma espermático a partir do T30. Conclui-se que as técnicas de recuperação de espermatozoides epididimários de cães castrados, testadas neste trabalho, podem ser utilizadas para refrigeração e criopreservação de sêmen.(AU)
The objective of this work was to evaluate the recovery of epididymal spermatozoa from castrated dogs using retrograde flow (FL) and flotation (FL) techniques in Tris-egg yolk diluent, before and after cryopreservation. Thirty testicle-epididymal complexes (CTE) were collected, 15 for FR and 15 for FL and soon after spermatozoid recovery, morphological changes in these spermatic cells were analyzed. After addition of the diluent, the parameters of total motility (MOT) and vigor (V) were evaluated. The post-cryopreserved semen was submitted to thermoresistance (TTR) test at T0, T30, T60 and T90 minutes, as well as the plasma and acrosomal membrane evaluation by fluorescent probes. There was no statistically significant difference between techniques tested for MOT and vigor in the diluted semen (FR-MOT: 82.3% and V: 3.4, FL-MOT: 79.6% and V: 3.2) and post-cryopreserved (FR-MOT: 34% and V: 2.8, FL-MOT: 30% and V: 2.7). From the T30 there was a significant difference regarding MOT and vigor in the used techniques, and the time also damaged the spermatic acrosome from the T30. It is concluded that the epididymal spermatozoa recovering techniques from castrated dogs, tested in this study, can be used for semen refrigeration and cryopreservation.(AU)
Subject(s)
Animals , Male , Dogs , Epididymis/physiology , Sperm Retrieval/veterinary , Semen Analysis/veterinary , Orchiectomy/veterinary , Cryopreservation/veterinaryABSTRACT
Abstract With the growing volume of research involving Thysanoptera in Brazil, studies were carried out to improve our understanding of the diversity of thrips in areas where the fauna has historically been neglected. Accordingly, we recorded the diversity of thrips (Insecta: Thysanoptera) associated with a restinga fragment located on the campus of the State University of Ceará (UECE), Fortaleza, Ceará state, and computed the estimated richness and diversity indices. Samples were collected from 2011 through 2013 from flowers of 86 plant species. The material was taken to the Laboratory of Insect-Plant Interaction, where thrips were screened under stereomicroscope. We collected 456 adults and 58 immatures, representing 14 species, in addition to one unidentified species of Treherniella. Microcephalothrips abdominalis was found on a large number of host plants, and Frankliniella insularis was the most common species. About two-thirds of the total richness of thrips species was associated with three plant families (Amaranthaceae, Caesalpiniaceae and Poaceae); six thrips species were each associated with only one plant species. The richness of the species collected was close to that estimated by Bootstrap and Jackknife 1 analysis. The Shannon-Wiener (H') and Simpson (D) diversity indexes were 1,7607 and 0.7769, respectively. Although the species are common, 46 new associations between plant species and thrips were established, 13 of which are true host associations, which demonstrates the importance of coastal vegetation in maintaining populations of thrips.
Resumo Com o crescente volume de pesquisas envolvendo Thysanoptera no Brasil, estudos foram promovidos para melhorar nosso entendimento da diversidade de tripes em área onde a fauna tem sido historicamente negligenciada. Dessa maneira, neste trabalho registramos a diversidade de tripes (Insecta: Thysanoptera) associada a um fragmento de restinga localizado no Campus da Universidade Estadual do Ceará (UECE), município de Fortaleza, estado do Ceará, e computamos a riqueza estimada de espécies e índices de diversidade. Foram realizadas coletas entre os anos de 2011 a 2013 em flores de 86 espécies vegetais. O material foi levado ao Laboratório de Interação Inseto-Planta (UECE) onde os tripes foram triados manualmente sob estereomicroscópio. Foram coletados 456 tripes adultos e 58 imaturos, representando 14 espécies, além de uma espécie não identificada de Treherniella. Microcephalothrips abdominalis foi encontrado em um maior número de plantas hospedeiras, e Frankliniella insularis foi a espécie mais frequente. Cerca de dois-terços da riqueza total de espécies de tripés está associada a três famílias botânicas (Amarnthaceae, Caesalpiniaceae e Poaceae); seis espécies de tripes foram encontradas em associação com somente uma espécie vegetal. A riqueza de espécies coletada foi próxima daquela estimada pelas análises Bootstrap e Jackknife 1. Os índices de diversidade de Shannon-Wiener (H') e Simpson (D) foram 1.7607 e 0,7769, respectivamente. Embora as espécies identificadas sejam comuns, 46 novas associações entre tripes e espécies vegetais foram constatadas, 13 das quais são associações hospedeiras verdadeiras, o que demonstra a importância da vegetação costeira na manutenção das populações de tripes.
Subject(s)
Animals , Forests , Flowers , Biodiversity , Thysanoptera/physiology , Brazil , Thysanoptera/classification , HerbivoryABSTRACT
With the growing volume of research involving Thysanoptera in Brazil, studies were carried out to improve our understanding of the diversity of thrips in areas where the fauna has historically been neglected. Accordingly, we recorded the diversity of thrips (Insecta: Thysanoptera) associated with a restinga fragment located on the campus of the State University of Ceará (UECE), Fortaleza, Ceará state, and computed the estimated richness and diversity indices. Samples were collected from 2011 through 2013 from flowers of 86 plant species. The material was taken to the Laboratory of Insect-Plant Interaction, where thrips were screened under stereomicroscope. We collected 456 adults and 58 immatures, representing 14 species, in addition to one unidentified species of Treherniella. Microcephalothrips abdominalis was found on a large number of host plants, and Frankliniella insularis was the most common species. About two-thirds of the total richness of thrips species was associated with three plant families (Amaranthaceae, Caesalpiniaceae and Poaceae); six thrips species were each associated with only one plant species. The richness of the species collected was close to that estimated by Bootstrap and Jackknife 1 analysis. The Shannon-Wiener (H') and Simpson (D) diversity indexes were 1,7607 and 0.7769, respectively. Although the species are common, 46 new associations between plant species and thrips were established, 13 of which are true host associations, which demonstrates the importance of coastal vegetation in maintaining populations of thrips.
Subject(s)
Biodiversity , Flowers , Forests , Thysanoptera/physiology , Animals , Brazil , Herbivory , Thysanoptera/classificationABSTRACT
Seminal plasma contains serine proteases and serine protease inhibitor, which are involved in mammalian fertilization, and the inhibitors can be applied to prevent cold-induced sperm capacitation. The effects of different concentrations of two serine protease inhibitors were analyzed, Plasminogen activator inhibitor 1 - PAI-1 (70Æg, 140Æg and 210 Æg) and Antipain (10µg, 50µg and 100µg) as supplementation to bovine semen cryopreservation extender. The effects of the inhibitors on the sperm parameters (sperm kinetics - CASA, acrosome integrity, plasma membrane integrity, mitochondrial membrane potential, sperm defects and acrosome reaction rate) were evaluated in the post-thaw semen. Cryopreservation of sperm with Antipain decreased post-thaw kinetic parameters of MP, VSL, LIN, SRT and the percentage of hyper-activated sperm while PAI-1 (210 Æg) decreased VSL and LIN. Antipain and PAI-1 had no effect on the integrity parameters of the plasma membrane, mitochondrial membrane potential and sperm defects. Sperm cryopreserved in the presence of Antipain and PAI-1 (70 and 140 Æg) preserved acrosome integrity, as they were able to complete the in vitro acrosome reaction. In conclusion, the serine protease inhibitors, Antipain and PAI-1 (70 and 140Æg) are able to preserve the acrosome integrity of cryopreserved bovine sperm.(AU)
A criopreservação é parcialmente prejudicial à fertilidade do sêmen de bovinos e induz mudanças semelhantes à capacitação em espermatozoides. O plasma seminal contém serina-proteases e inibidores de serina-proteases que estão envolvidos na fertilização de mamíferos, e os inibidores podem ser aplicados para evitar uma capacitação espermática induzida pelo frio. Analisaram-se os efeitos de diferentes concentrações de dois inibidores de serina-proteases, inibidor do ativador do plasminogênio 1 - PAI-1 (70Æg, 140Æg e 210Æg) e antipaína (10µg, 50µg e 100µg) na suplementação ao diluidor de criopreservação de sêmen bovino. Trinta e seis ejaculados de quatro bovinos Curraleiro Pé-Duro foram usados para criopreservação. Os efeitos dos inibidores sobre os parâmetros dos espermatozoides (cinética espermática - CASA, integridade acrossomal, integridade da membrana plasmática, potencial de membrana mitocondrial, defeitos espermáticos e taxa de reação acrossomal) foram avaliados no sêmen pós-descongelamento. A criopreservação de espermatozoides com antipaína diminuiu os parâmetros cinéticos pós-descongelamento de MP, VSL, LIN, SRT e a porcentagem de espermatozoides hiperativados, PAI-1 (210Æg) diminuiu VSL e LIN. Antipaína e PAI-1 não tiveram efeitos nos parâmetros de integridade da membrana plasmática, no potencial de membrana mitocondrial e nos defeitos espermáticos. Espermatozoides criopreservados na presença de antipaína e PAI-1 (70 e 140Æg) preservaram a integridade acrossomal, assim como foram capazes de completar a reação acrossômica in vitro. Em conclusão, os inibidores de serina-proteases, antipaína e PAI-1 (70 e 140Æg) são capazes de preservar a integridade acrossomal de espermatozoides criopreservados de bovinos.(AU)
Subject(s)
Animals , Male , Cattle , Acrosome , Antipain/antagonists & inhibitors , Cryopreservation/veterinary , Plasminogen Activators/antagonists & inhibitors , Serine Proteinase Inhibitors/analysis , Cryopreservation/methods , Semen Analysis/veterinary , Semen Preservation/veterinaryABSTRACT
The success of the geographical distribution of goat populations around the world is a consequence of the adaptive potential of these breeds. Several relevant traits to the success of the species in colonizing different ecosystems (and use by man) evolved before domestication. These features were relevant for the selection of different breeds. Each breed represents a genetic heritage that may be unique and essential for maintaining the species. The objective of this study was to catalog the mtDNA haplotypes of the Brazilian autochthonous Canindé goat breed and to characterize the genetic diversity observed in subpopulations by sequencing a 481-bp fragment corresponding to the first portion of the control region in 178 individuals from 10 herds, sampled in six Brazilian states. The global population displays a total of 29 haplotypes and 56 polymorphic sites. About one-third (10) of the haplotypes were common to all subpopulations while the remaining (19) were exclusive to a single subpopulation. The population exhibited high average haplotype diversity (0.82), with maximum and minimum values of 0.90 and 0.56 in individual subpopulations, respectively. In contrast, nucleotide diversity was 0.014, with maximum and minimum values of 0.020 and 0.004, respectively. The spatial analysis of molecular variance did not detect structure within the Canindé goat breed, and analysis of molecular variance revealed that 88.4% of the variation observed in the population was due to differences among individuals in the same subpopulation. Only 11.4% of the genetic variation referred to differences among subpopulations. About one-third (33.1%) of the individuals within population shared the same haplotype, which may be due not only to the breed developing from a small number of matrilines. The Brazilian autochthonous Canindé breed was classified as haplogroup A, a haplotype predominant in the Europe region.
Subject(s)
DNA, Mitochondrial/genetics , Goats/genetics , Polymorphism, Genetic , Animals , HaplotypesABSTRACT
Autosomal dominant polycystic kidney disease (ADPKD) is a hereditary nephropathy characterized by abnormal growth of epithelial cells. Genetic factors, including the vascular endothelial growth factor (VEGF) gene, play an important role in its progression. The main aim of this study was to evaluate the influence of VEGF-C936T polymorphism in the development and progression of ADPKD. In total, 302 individuals were studied and divided into two groups: G1 (73 patients with ADPKD) and G2 (229 individuals without the disease). Among the patients, 46 (63%) progressed to end-stage renal disease (ESRD), and required hemodialysis and/or renal transplant. These patients were re-grouped into G1-A for progression analysis. A peripheral blood sample was obtained from all subjects; the DNA was extracted and the VEGF-C936T polymorphism analyzed using polymerase chain reaction/restriction fragment length polymorphism. The significance level was set at P < 0.05. The homozygous wild-type genotype (C/C) was predominant in G1 (78%) and G2 (79%; P = 0.9249). We observed a significant reduction in the mean age of patients with the risk allele (C/T + T/T = 44.3 ± 13.4 years) compared to the C/C genotype (52.2 ± 9.6 years; P = 0.047) in G1-A. In conclusion, the VEGF-C936T polymorphism does not discriminate patients from controls. However, the presence of the T allele appears to accelerate the progression of ADPKD, anticipating ESRD, thereby suggesting its importance in the prognosis of the disease. However, the importance role played by VEGF gene variants in different populations and larger sample sizes must be verified.
Subject(s)
Kidney Failure, Chronic/genetics , Neovascularization, Pathologic/genetics , Polycystic Kidney, Autosomal Dominant/genetics , Polymorphism, Single Nucleotide , Vascular Endothelial Growth Factor A/genetics , Adult , Age Factors , Alleles , Biomarkers/metabolism , Case-Control Studies , Cell Proliferation , Disease Progression , Epithelial Cells/metabolism , Epithelial Cells/pathology , Female , Gene Expression , Gene Frequency , Genotype , Homozygote , Humans , Kidney/metabolism , Kidney/pathology , Kidney Failure, Chronic/diagnosis , Kidney Failure, Chronic/etiology , Kidney Failure, Chronic/pathology , Male , Neovascularization, Pathologic/complications , Neovascularization, Pathologic/diagnosis , Neovascularization, Pathologic/pathology , Polycystic Kidney, Autosomal Dominant/complications , Polycystic Kidney, Autosomal Dominant/diagnosis , Polycystic Kidney, Autosomal Dominant/pathology , Risk , Vascular Endothelial Growth Factor A/metabolismABSTRACT
Successful DNA extraction is indispensable for molecular methods based on polymerase chain reaction (PCR); however, goat sperm DNA extraction is limited. Thus, the aim of this study was to evaluate three methods to extract DNA from goat sperm for use in PCR. Eight goat semen pools were used for DNA extraction by using the DNeasy Blood & Tissue Kit, phenol-chloroform, and Chelex-100 methods. DNA samples were analyzed spectrophotometrically to determine the DNA concentration and purity, visualized on 0.8% agarose gel, and used at different amounts (150, 100, 50, 10, and 1 ng) for PCR with electrophoresis, followed by 1.5% agarose gel electrophoresis. The quantity of DNA extracted with Chelex-100 was higher (P < 0.05) than that obtained with either the DNeasy Blood & Tissue Kit or the phenol-chloroform method, with the phenol-chloroform method yielding a greater quantity (P < 0.05) than the kit. The DNeasy Blood & Tissue Kit produced a higher (P < 0.05) purity product than the Chelex-100 method, and all samples obtained by the three protocols were positive for DNA, as assessed by electrophoresis. All of the different concentrations of DNA produced by these methods were amplified by PCR, although for DNA produced by the phenol-chloroform method, PCR was only possible after complementary purification. In conclusion, the Chelex-100 method is cheap, secure, simple, fast, and effective, and is a potential tool for extracting goat sperm DNA without limitations in PCR.
Subject(s)
DNA/isolation & purification , Polymerase Chain Reaction , Spermatozoa/metabolism , Animals , Goats , Male , SemenABSTRACT
A expressão de RNAm para leptina, receptor de leptina (obRb), adiponectina, receptor de adiponectina (AdipoR1) e resistina foi avaliada por meio da técnica de PCR em tempo real, em tecidos ovariano, hipofisário, adiposo do omento e da região perirrenal, em ovelhas alimentadas sem farelo de mamona ou com farelo de mamona detoxificada durante 14 meses. O tipo de dieta não afetou os níveis de RNAm para leptina, obRb, adiponectina, AdipoR1 e resistina nos diferentes tecidos avaliados (P>0,05). Nos tecidos ovariano e hipofisário, não foi verificada a expressão da adiponecina e da resistina, respectivamente. Como consequência, pode-se concluir que o farelo de mamona detoxificada pode ser utilizado como fonte proteica na dieta de ovelhas, sem afetar a expressão do gene resistina e dos genes leptina e adiponectina, bem como de seus receptores...
The expression of leptin, leptin receptor (obRb), adiponectin, adiponectin receptor (AdipoR1) and resistin was assessed by real-time PCR technique in ovarian, pituitary, and the omental adipose perirenal tissue in sheep feed without castor meal or with detoxified castor meal. The type of diet did not affect mRNA levels for leptin, obRb, adiponectin, resistin AdipoR1 evaluated in different tissues (P>0.05). However, in pituitary and ovarian tissues there was no expression of resistin and adiponectin, respectively. The detoxified castor meal can be used in sheep diets as alternative food protein without affecting the expression of leptin and adponectin as well as their receptors and resistin...
Subject(s)
Animals , Sheep/metabolism , Receptors, Adipokine/analysis , Receptors, Leptin/analysis , Reproduction/physiology , Resistin/analysis , Animal Feed , Ricinus , Polymerase Chain Reaction/veterinaryABSTRACT
Planned conversion from tacrolimus to sirolimus was evaluated in de novo kidney transplant recipients. In this multicenter, randomized, open-label study, 297 patients were initially treated with tacrolimus, mycophenolate sodium and prednisone. Of the 283 patients reaching 3 months, 97 were converted to sirolimus (SRL), 107 were maintained on tacrolimus (TAC) and 79 were patients receiving TAC without criteria to undergo intervention at month 3 (TACex). The primary objective was to show superior estimated glomerular filtration rate (eGFR) in the SRL group at month 24. Of the 258 patients who completed 24 months, 91 (94%) were in the SRL group, 101 (94%) in the TAC group and 66 (84%) in the TACex group. In the intention-to-treat population there were no differences in eGFR (66.2 ± 25.3 vs. 70.7 ± 25.1, p = 0.817) or in the severity of chronic sclerosing lesions scores in 24-month protocol biopsies. Higher mean urinary protein-to-creatinine ratio (0.36 ± 0.69 vs. 0.15 ± 0.53, p = 0.03) and higher incidence of treated acute rejection between months 3-24 (13.4% vs. 4.7%, p = 0.047) were observed in SRL compared to TAC group. In this population planned conversion from TAC to SRL 3 months after kidney transplantation was not associated with improved renal function at 24 months.
Subject(s)
Graft Rejection/prevention & control , Kidney Transplantation , Renal Insufficiency/therapy , Sirolimus/administration & dosage , Tacrolimus/administration & dosage , Adult , Biopsy , Creatinine/blood , Female , Glomerular Filtration Rate , Humans , Immunosuppressive Agents/administration & dosage , Immunosuppressive Agents/adverse effects , Male , Middle Aged , Mycophenolic Acid/administration & dosage , Mycophenolic Acid/analogs & derivatives , Prednisone/administration & dosage , Prospective Studies , Sirolimus/adverse effects , Tacrolimus/adverse effects , Treatment OutcomeABSTRACT
Leptin, a hormone that was originally identified in adipocytes, has been implicated in the regulation of ovarian folliculogenesis through endocrine, autocrine and/or paracrine mechanisms. The aim of this study was to investigate the expression patterns of leptin (LEP) and its receptor (LEPRb) in different types of ovarian follicular cells from goats. In small follicles, the expression levels of LEP were higher (P<0.001) in granulosa cells than in theca cells, cumulus cells and oocytes. The expression of LEP in granulosa cells was higher (P<0.001) in small follicles than in large follicles. In large follicles, the expression of LEPRb was higher (P<0.05) in granulosa cells than in theca cells, cumulus cells and oocytes. Higher expression (P<0.05) of LEPRb was detected in granulosa cells isolated from large follicles than in granulosa cells isolated from small follicles. Immunohistochemical analyses revealed the presence of the LEP and LEPR proteins in follicles at all stages of development. The most intense staining for LEP and LEPR was observed in the cytoplasm of oocytes and the surrounding granulosa cells. In conclusion, it was demonstrated that leptin and its receptor are expressed at both the mRNA and protein levels in goat ovarian follicles. Furthermore, the presence of a leptin signaling system in the caprine ovary suggests a potential regulatory role for leptin in follicular development and the maturation of goat oocytes.
Subject(s)
Gene Expression Regulation/physiology , Goats/metabolism , Leptin/metabolism , Ovarian Follicle/metabolism , Receptors, Leptin/metabolism , Animals , Female , Leptin/genetics , Ovarian Follicle/cytology , Protein Transport , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptors, Leptin/genetics , Signal TransductionABSTRACT
Abscisic acid is a plant hormone that participates in essential plant physiological processes, especially during adaptation to many environmental stresses, such as water deficit. The relationship between ABA accumulation and the expression of putative carotenoid cleavage dioxygenase (CCD) genes was investigated in the pot-cultivated leaves and roots of the 'Rangpur' lime and 'Sunki Maravilha' mandarin plants. Transpiration, stomatal resistance and leaf growth were evaluated when these genotypes were subjected to continuous water deficit. Under water deficit conditions, the 'Rangpur' lime extracts used greater amounts of water when compared to the 'Sunki Maravilha' plants, which reached the greatest stomatal resistance 5 days before 'Rangpur' lime. When subjected to water deficit, the roots and leaves of 'Sunki Maravilha' showed a progressive increase in ABA accumulation; however, in 'Rangpur' lime, alternations between high and low ABA concentrations were observed. These results suggest a retroactive feeding regulation by ABA. In 'Rangpur' lime the NCED2, NCED3 and CCD4a genes were expressed at the highest levels in the roots, and NCED5 was highly expressed in the leaves; in 'Sunki Maravilha', the NCED2 and NCED5 genes were most highly expressed in the roots, and NCED2 was most highly expressed in the leaves. However, for both genotypes, the transcription of these genes only correlated with ABA accumulation during the most severe water deficit conditions. The 'Rangpur' lime behaved as a vigorous rootstock; the leaf growth remained unaltered even when water was scarce. However, 'Sunki Maravilha' adaptation was based on the equilibrium of the response between the root and the aerial tissues due to water restriction. The use of the Sunki mandarin in combination with a scion with similar characteristics as its own, which responds to water deficit stress by accumulating ABA in the leaves, may display good drought tolerance under field conditions.
Subject(s)
Abscisic Acid/metabolism , Adaptation, Biological/genetics , Citrus/physiology , Dioxygenases/metabolism , Droughts , Gene Expression Regulation, Plant/physiology , Plant Proteins/metabolism , Agriculture/methods , Citrus/enzymology , Citrus/metabolism , DNA Primers/genetics , Dioxygenases/genetics , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Roots/metabolism , Real-Time Polymerase Chain Reaction , Species Specificity , Water DeprivationABSTRACT
The molecular integration of nutrient- and pathogen-sensing pathways has become of great interest in understanding the mechanisms of insulin resistance in obesity. The double-stranded RNA-dependent protein kinase (PKR) is one candidate molecule that may provide cross talk between inflammatory and metabolic signaling. The present study was performed to determine, first, the role of PKR in modulating insulin action and glucose metabolism in physiological situations, and second, the role of PKR in insulin resistance in obese mice. We used Pkr(-/-) and Pkr(+/+) mice to investigate the role of PKR in modulating insulin sensitivity, glucose metabolism, and insulin signaling in liver, muscle, and adipose tissue in response to a high-fat diet. Our data show that in lean Pkr(-/-) mice, there is an improvement in insulin sensitivity, and in glucose tolerance, and a reduction in fasting blood glucose, probably related to a decrease in protein phosphatase 2A activity and a parallel increase in insulin-induced thymoma viral oncogene-1 (Akt) phosphorylation. PKR is activated in tissues of obese mice and can induce insulin resistance by directly binding to and inducing insulin receptor substrate (IRS)-1 serine307 phosphorylation or indirectly through modulation of c-Jun N-terminal kinase and inhibitor of κB kinase ß. Pkr(-/-) mice were protected from high-fat diet-induced insulin resistance and glucose intolerance and showed improved insulin signaling associated with a reduction in c-Jun N-terminal kinase and inhibitor of κB kinase ß phosphorylation in insulin-sensitive tissues. PKR may have a role in insulin sensitivity under normal physiological conditions, probably by modulating protein phosphatase 2A activity and serine-threonine kinase phosphorylation, and certainly, this kinase may represent a central mechanism for the integration of pathogen response and innate immunity with insulin action and metabolic pathways that are critical in obesity.
Subject(s)
Insulin Resistance/physiology , Obesity/metabolism , RNA, Double-Stranded/metabolism , eIF-2 Kinase/metabolism , Animals , Blood Glucose/genetics , Blood Glucose/metabolism , Eating/physiology , Glucose/metabolism , Glucose Intolerance/genetics , Glucose Intolerance/metabolism , Insulin Receptor Substrate Proteins/genetics , Insulin Receptor Substrate Proteins/metabolism , Liver/drug effects , Liver/metabolism , Macrophages/drug effects , Macrophages/metabolism , Mice , Mice, Knockout , Muscle, Skeletal/drug effects , Muscle, Skeletal/metabolism , Obesity/genetics , Oxygen Consumption/physiology , Palmitic Acid/pharmacology , Phosphorylation , Protein Phosphatase 2/metabolism , Proto-Oncogene Proteins c-akt/metabolism , RNA, Double-Stranded/genetics , Signal Transduction/drug effects , eIF-2 Kinase/geneticsABSTRACT
Nodular glomerulosclerosis may be idiopathic or develop associated with diabetes mellitus, membranoprolipherative glomerulonephritis, light or heavy chain deposits, amyloidosis, fibrillary or immunotactoide disease, and Takayasu's arteritis. Histological features of idiopathic nodular glomerulosclerosis are similar to the Kimmelstiel-Wilson changes. Recent evidence points to the role of hyperglycemia, hyperlipidemia, hypertension and smoking in the mechanisms of this uncommon condition. The case study of a 65-year-old male presenting recent arterial hypertension and nodular non-diabetic glomerulosclerosis is described, and the possible role of heavy smoking in the pathogenesis of this condition is emphasized.
Subject(s)
Diabetic Nephropathies/etiology , Dyslipidemias/complications , Hypertension/complications , Smoking/adverse effects , Humans , Male , Middle AgedABSTRACT
The aim of this study was to characterize the reproductive disorders in the acute and chronic phases in ewes experimentally infected with different doses of Toxoplasma gondii during artificial insemination occurred. Animals (n=41) were divided into three experimental groups: in the group 1 (G1, n=15), animals were inseminated using contaminated semen containing 6.5×104 tachyzoites; in the group 2 (G2, n=15), animals were inseminated with contaminated semen containing 4×107 tachyzoites and in the group 3 (G3, n=11), animals were inseminated using tachyzoite-free semen, serving as control group. Parasitemia and seroconversion were observed in 28 of 30 and 20 of 30, respectively, from the seventh day after infection. Embryonic reabsorption was observed in the acute phase in ewes from G1 and G2. Persistent anestrus, hydrometra, mucometra and follicular cysts were observed in the second phase of the experiment in animals from G1 and G2. Histopathological lesions similar to those of toxoplasmosis were found in the placentas. In conclusion, artificial insemination using semen containing experimentally added tachyzoites can establish toxoplasmosis in ewes and cause reproductive pathologies during the acute and chronic phases of the disease.
Subject(s)
Follicular Cyst/veterinary , Insemination, Artificial/veterinary , Placenta Diseases/veterinary , Pregnancy, Animal , Semen/parasitology , Sheep Diseases/parasitology , Toxoplasmosis, Animal/complications , Uterine Diseases/veterinary , Acute Disease , Anestrus , Animals , Chronic Disease , Female , Follicular Cyst/parasitology , Follicular Cyst/pathology , Placenta Diseases/parasitology , Placenta Diseases/pathology , Pregnancy , Toxoplasmosis, Animal/pathology , Uterine Diseases/parasitology , Uterine Diseases/pathologyABSTRACT
AIM/HYPOTHESIS: High-dose aspirin treatment improves fasting and postprandial hyperglycaemia in patients with type 2 diabetes, as well as in animal models of insulin resistance associated with obesity and sepsis. In this study, we investigated the effects of aspirin treatment on inducible nitric oxide synthase (iNOS)-mediated insulin resistance and on S-nitrosylation of insulin receptor (IR)-beta, IRS-1 and protein kinase B (Akt) in the muscle of diet-induced obese rats and also in iNos (also known as Nos2)-/- mice on high fat diet. METHODS: Aspirin (120 mg kg-1 day-1 for 2 days) or iNOS inhibitor (L-NIL; 80 mg/kg body weight) were administered to diet-induced obese rats or mice and iNOS production and insulin signalling were investigated. S-nitrosylation of IRbeta/IRS-1 and Akt was investigated using the biotin switch method. RESULTS: iNOS protein levels increased in the muscle of diet-induced obese rats, associated with an increase in S-nitrosylation of IRbeta, IRS-1 and Akt. These alterations were reversed by aspirin treatment, in parallel with an improvement in insulin signalling and sensitivity, as measured by insulin tolerance test and glucose clamp. Conversely, while aspirin reversed the increased phosphorylation of IkappaB kinase beta and c-Jun amino-terminal kinase, as well as IRS-1 serine phosphorylation in diet-induced obese rats and iNos -/- mice on high-fat diet, these alterations were not associated with the improvement of insulin action induced by this drug. CONCLUSIONS/INTERPRETATION: Our data demonstrate that aspirin treatment not only reduces iNOS protein levels, but also S-nitrosylation of IRbeta, IRS-1 and Akt. These changes are associated with improved insulin resistance and signalling, suggesting a novel mechanism of insulin sensitisation evoked by aspirin treatment.
Subject(s)
Aspirin/therapeutic use , Insulin Resistance/physiology , Muscle, Skeletal/physiopathology , Nitric Oxide Synthase Type II/antagonists & inhibitors , Obesity/physiopathology , Animals , Drug Tolerance/physiology , Insulin/physiology , Insulin Receptor Substrate Proteins/metabolism , Male , Mice , Mice, Inbred C57BL , Mice, Obese , Muscle, Skeletal/drug effects , Muscle, Skeletal/enzymology , Nitric Oxide Synthase/metabolism , Nitric Oxide Synthase Type II/biosynthesis , Proto-Oncogene Proteins c-akt/metabolism , Rats , Rats, WistarABSTRACT
Type 2 diabetes increases the risk of cardiovascular mortality and these patients, even without previous myocardial infarction, run the risk of fatal coronary heart disease similar to non-diabetic patients surviving myocardial infarction. There is evidence showing that particulate matter air pollution is associated with increases in cardiopulmonary morbidity and mortality. The present study was carried out to evaluate the effect of diabetes mellitus on the association of air pollution with cardiovascular emergency room visits in a tertiary referral hospital in the city of São Paulo. Using a time-series approach, and adopting generalized linear Poisson regression models, we assessed the effect of daily variations in PM10, CO, NO2, SO2, and O3 on the daily number of emergency room visits for cardiovascular diseases in diabetic and non-diabetic patients from 2001 to 2003. A semi-parametric smoother (natural spline) was adopted to control long-term trends, linear term seasonal usage and weather variables. In this period, 45,000 cardiovascular emergency room visits were registered. The observed increase in interquartile range within the 2-day moving average of 8.0 microg/m(3) SO2 was associated with 7.0% (95%CI: 4.0-11.0) and 20.0% (95%CI: 5.0-44.0) increases in cardiovascular disease emergency room visits by non-diabetic and diabetic groups, respectively. These data indicate that air pollution causes an increase of cardiovascular emergency room visits, and that diabetic patients are extremely susceptible to the adverse effects of air pollution on their health conditions.
Subject(s)
Air Pollutants/adverse effects , Air Pollution/adverse effects , Cardiovascular Diseases/etiology , Diabetes Mellitus , Emergency Service, Hospital/statistics & numerical data , Air Pollutants/classification , Air Pollution/statistics & numerical data , Brazil/epidemiology , Cardiovascular Diseases/epidemiology , Diabetes Mellitus/epidemiology , Humans , Particulate Matter/toxicity , Poisson DistributionABSTRACT
Type 2 diabetes increases the risk of cardiovascular mortality and these patients, even without previous myocardial infarction, run the risk of fatal coronary heart disease similar to non-diabetic patients surviving myocardial infarction. There is evidence showing that particulate matter air pollution is associated with increases in cardiopulmonary morbidity and mortality. The present study was carried out to evaluate the effect of diabetes mellitus on the association of air pollution with cardiovascular emergency room visits in a tertiary referral hospital in the city of São Paulo. Using a time-series approach, and adopting generalized linear Poisson regression models, we assessed the effect of daily variations in PM10, CO, NO2, SO2, and O3 on the daily number of emergency room visits for cardiovascular diseases in diabetic and non-diabetic patients from 2001 to 2003. A semi-parametric smoother (natural spline) was adopted to control long-term trends, linear term seasonal usage and weather variables. In this period, 45,000 cardiovascular emergency room visits were registered. The observed increase in interquartile range within the 2-day moving average of 8.0 µg/m³ SO2 was associated with 7.0 percent (95 percentCI: 4.0-11.0) and 20.0 percent (95 percentCI: 5.0-44.0) increases in cardiovascular disease emergency room visits by non-diabetic and diabetic groups, respectively. These data indicate that air pollution causes an increase of cardiovascular emergency room visits, and that diabetic patients are extremely susceptible to the adverse effects of air pollution on their health conditions.
Subject(s)
Humans , Air Pollutants/adverse effects , Air Pollution/adverse effects , Cardiovascular Diseases/etiology , Diabetes Mellitus , Emergency Service, Hospital/statistics & numerical data , Air Pollutants/classification , Air Pollution/statistics & numerical data , Brazil/epidemiology , Cardiovascular Diseases/epidemiology , Diabetes Mellitus/epidemiology , Poisson Distribution , Particulate Matter/toxicityABSTRACT
Microdeletions in Yq are associated with defects in spermatogenesis, while those in the AZF region are considered critical for germ cell development. We examined microdeletions in the Y chromosomes of patients attended at the Laboratory of Human Reproduction of the Clinical Hospital of the Federal University of Goiás as part of a screening of patients who plan to undergo assisted reproduction. Analysis was made of the AZF region of the Y chromosome in men who had altered spermograms to detect possible microdeletions in Yq. Twenty-three patients with azoospermia and 40 with severe oligozoospermia were analyzed by PCR for the detection of six sequence-tagged sites: sY84 and sY86 for AZFa, sY127 and sY134 for AZFb, and sY254 and sY255 for AZFc. Microdeletions were detected in 28 patients, including 10 azoospermics and 18 severe oligozoospermics. The patients with azoospermia had 43.4% of their microdeletions in the AZFa region, 8.6% in the AZFb region and 17.4% in the AZFc region. In the severe oligozoospermics, 40% were in the AZFa region, 5% in the AZFb region and 5% in the AZFc region. We conclude that microdeletions can be the cause of idiopathic male infertility, supporting conclusions from previous studies.
Subject(s)
Chromosome Deletion , Chromosomes, Human, Y/ultrastructure , Gene Deletion , Infertility, Male/genetics , Azoospermia/genetics , Brazil , Fertility , Germ Cells/metabolism , Humans , Male , Polymerase Chain Reaction , SpermatogenesisABSTRACT
Microdeletions in Yq are associated with defects in spermatogenesis, while those in the AZF region are considered critical for germ cell development. We examined microdeletions in the Y chromosomes of patients attended at the Laboratory of Human Reproduction of the Clinical Hospital of the Federal University of Goiás as part of a screening of patients who plan to undergo assisted reproduction. Analysis was made of the AZF region of the Y chromosome in men who had altered spermograms to detect possible microdeletions in Yq. Twenty-three patients with azoospermia and 40 with severe oligozoospermia were analyzed by PCR for the detection of six sequence-tagged sites: sY84 and sY86 for AZFa, sY127 and sY134 for AZFb, and sY254 and sY255 for AZFc. Microdeletions were detected in 28 patients, including 10 azoospermics and 18 severe oligozoospermics. The patients with azoospermia had 43.4% of their microdeletions in the AZFa region, 8.6% in the AZFb region and 17.4% in the AZFc region. In the severe oligozoospermics, 40% were in the AZFa region, 5% in the AZFb region and 5% in the AZFc region. We conclude that microdeletions can be the cause of idiopathic male infertility, supporting conclusions from previous studies.
