RESUMO
In this study, we investigated the mitochondrial quality control system in porcine oocytes during meiotic maturation. Cumulus cell oocyte complexes (COCs) collected from gilt ovaries were treated with 10â µM carbonyl cyanide-m-chlorophenylhydrazone (CCCP; a mitochondrial uncoupler) for 2 âh. The CCCP treatment was found to significantly reduce ATP content, increase the amount of phosphorylated AMP-activated protein kinase and elevate reactive oxygen species levels in oocytes. When the CCCP-treated COCs were cultured further for 44â h in maturation medium, the ATP levels were restored and the parthenogenetic developmental rate of oocytes to the blastocyst stage was comparable with that of untreated COCs. To examine the effects of CCCP treatment of oocytes on the kinetics of mitochondrial DNA copy number (Mt number), COCs treated with 0 or 10 âµM CCCP were cultured for 44 âh, after which the Mt number was determined by RT-PCR. CCCP treatment was found to increase the Mt number in the modified maturation medium in which mitochondrial degradation was inhibited by MG132, whereas CCCP treatment did not affect the Mt number in the maturation medium lacking MG132. The relative gene expression of TFAM was furthermore shown to be significantly higher in CCCP-treated oocytes than in untreated oocytes. Taken together, the finding presented here suggest that when the mitochondria are injured, mitochondrial biogenesis and degradation are induced, and that these processes may contribute to the recuperation of oocytes.
Assuntos
Carbonil Cianeto m-Clorofenil Hidrazona/toxicidade , Mitocôndrias/efeitos dos fármacos , Oócitos/efeitos dos fármacos , Biogênese de Organelas , Desacopladores/toxicidade , Proteínas Quinases Ativadas por AMP/metabolismo , Trifosfato de Adenosina/metabolismo , Animais , Células do Cúmulo/efeitos dos fármacos , Feminino , Dosagem de Genes , Expressão Gênica/efeitos dos fármacos , Técnicas In Vitro , Mitocôndrias/metabolismo , Partenogênese/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , SuínosRESUMO
During oocyte growth, the number of mitochondria drastically increases and mitochondrial function profoundly affects the oocyte competence. Resveratrol is a well-known activator of sirtuin 1 (SIRT1), which has a role in cellular energy homeostasis and mitochondrial biogenesis. The main aim of the present study was to examine the effect of supplementation of culture media with resveratrol on oocyte development and mitochondrial number and functions. Lipid contents and developmental ability of the oocytes grown in vitro were also examined. Oocyte-granulosa cell complexes were collected from early antral follicles of gilt ovaries and were cultured in medium containing 0 or 2 µM resveratrol for 14 days. Immunostaining revealed that resveratrol enhanced SIRT1 expression in oocytes. Antrum formation during the culture period and survivability of the granulosa cells surrounding the developed oocytes did not differ between the two concentrations of resveratrol. In addition, the ability of oocytes to complete meiotic maturation did not differ between the two concentrations of resveratrol, whereas the ability of oocytes to develop to the blastocyst stage was improved significantly by resveratrol (7.4% vs. 1.6%; P < 0.05). Resveratrol upregulated the ATP content in oocytes grown in vitro, and the addition of 2 µM of the SIRT1 inhibitor 6-Chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (EX527) diminished this effect although EX527 alone had no effect on ATP content. The mitochondrial DNA copy number in oocytes determined by quantitative real-time polymerase chain reaction increased during in vitro oocyte development, but resveratrol did not affect the kinetics of the mitochondrial DNA copy number. We found that resveratrol also increased the expression level of phospho-5'-adenosine monophosphate-activated protein kinase in oocytes but decreased the lipid content in oocytes grown in vitro. These results suggest that resveratrol increased the ATP content in oocytes via energy homeostasis and improved the developmental ability of oocytes grown in vitro.
Assuntos
Técnicas de Maturação in Vitro de Oócitos/métodos , Oócitos/fisiologia , Sirtuínas/metabolismo , Estilbenos/administração & dosagem , Sus scrofa , Proteínas Quinases Ativadas por AMP/análise , Trifosfato de Adenosina/análise , Animais , Meios de Cultura , DNA Mitocondrial/análise , Metabolismo Energético/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Feminino , Células da Granulosa/efeitos dos fármacos , Lipídeos/análise , Oócitos/química , Oócitos/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Resveratrol , Sirtuínas/análiseRESUMO
PURPOSE: To assess the age-associated changes in oocytes and granulosa cells derived from early antral follicles (EAFs). METHOD: Gene expression analysis of granulosa cells of the EAFs using a genome analyzer (Illumina) and in vitro culture of oocyte-granulosa cell complexes (OGCs) of EAFs (400-700 µm in diameter) collected from ovaries of aged (>120 months) and young (<50 months) cows. RESULTS: Gene expression profiles in granulosa cells of EAFs of aged cows, which included changes in genes that encode chaperone proteins and antioxidants. In vivo development of EAFs, as determined by oocyte diameter of EAFs and AFs (3-6 mm in diameter), appeared to be impaired in aged cows and the OGCs of aged cows contained low GSH compared to younger counterparts. When the OGCs were cultured in a medium containing low estradiol (E2, 0.1 µg/mL), the ratio of antrum formation was higher for OGCs from aged animals than that from young animals, while higher abnormal fertilization rate and lower total cell number of the blastocysts were observed in the OGCs of aged cows compared with those of young cows. On the contrary, when the OGCs were cultured in a medium containing 10 µg/mL E2, the ratio of antrum formation and fertilization outcome was comparable between the two age groups, whereas the total cell number of the blastocysts was still low in the aged group. CONCLUSION: Aging affects the gene expression profiles of the granulosa cells, and impairs in vitro developmental ability of OGCs collected from EAFs.
Assuntos
Embrião de Mamíferos/citologia , Células da Granulosa/citologia , Oócitos/citologia , Folículo Ovariano/citologia , Fatores Etários , Animais , Biomarcadores/metabolismo , Bovinos , Técnicas de Cultura de Células , Embrião de Mamíferos/efeitos dos fármacos , Embrião de Mamíferos/metabolismo , Estradiol/farmacologia , Feminino , Fertilização , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Células da Granulosa/efeitos dos fármacos , Células da Granulosa/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismoRESUMO
Because the contribution of residual renal function (RRF) to total solute clearance is often significant in continuous ambulatory peritoneal dialysis (CAPD), loss of RRF over time can lead to inadequate dialysis if appropriate prescription management strategies are not pursued. Additionally, declines in ultrafiltration caused by increases in peritoneal permeability may limit continuation of CAPD therapy. Peritoneal dialysis and hemodialysis (PD + HD) combination therapy (complementary dialysis therapy) is an alternative method. This therapy allows the patient to maintain daily activities, as with CAPD, while undergoing once-a-week HD supplements for the insufficient removal of solutes and water. This therapy allows for the continuation of PD without shifting to total HD in PD patients who continue to have uremic symptoms even after individualization of the PD prescription. This treatment option is psychologically more acceptable to patients and may be expected to provide such accompanying beneficial effects as peritoneal resting, improvement of QOL and reduction in medical cost.
Assuntos
Falência Renal Crônica/terapia , Diálise Peritoneal Ambulatorial Contínua/métodos , Diálise Renal/métodos , Terapia Combinada , Humanos , Diálise Peritoneal Ambulatorial Contínua/economia , Diálise Peritoneal Ambulatorial Contínua/normas , Qualidade de Vida , Diálise Renal/economia , Diálise Renal/normasRESUMO
The aim of this study was to define the incidence and characteristics of sclerosing encapsulating peritonitis (SEP) in pediatric peritoneal dialysis (PD) patients in Japan. A questionnaire was sent to all dialysis units with at least two pediatric PD patients. Among 687 patients registered, 11 cases (1.6%) of SEP were diagnosed. The mean age of patients with SEP at the start of PD was 9.7+/-3.6 years and at SEP diagnosis, 19.1+/-3.8 years. All patients had undergone PD for more than 5 years, and the mean PD duration was 9.6+/-3.3 years. SEP was diagnosed in 6.6% and 12% of patients dialyzed for >5 years and >8 years, respectively. The incidence of peritonitis among patients with SEP was not different from that among the Japanese pediatric registry. All patients had virtually no residual urine volume and 9 had impaired peritoneal ultrafiltration. Peritoneal calcification was the most-frequent radiological finding. Peritoneal biopsy was performed in 7 patients and confirmed sclerotic peritonitis in all. Ten patients transferred to hemodialysis, and only 1 patient underwent surgery. Three patients died. We recommend that patients on PD for more than 5 years who have impaired peritoneal ultrafiltration or peritoneal calcification should be carefully managed as presumptive cases of SEP.
Assuntos
Diálise Peritoneal/efeitos adversos , Peritonite/etiologia , Peritonite/patologia , Adolescente , Adulto , Candidíase , Enterococcus , Infecções por Bactérias Gram-Positivas , Humanos , Mortalidade , Peritônio/patologia , Peritonite/diagnóstico , Peritonite/microbiologia , Esclerose , Infecções EstafilocócicasRESUMO
We performed MR imaging of the brain with both conventional spin-echo (SE) and fluid-attenuated inversion-recovery (FLAIR) sequences in a case of Wernicke encephalopathy. Lesion conspicuity was found to be better with FLAIR imaging.
Assuntos
Encéfalo/patologia , Imageamento por Ressonância Magnética/métodos , Encefalopatia de Wernicke/diagnóstico , Artefatos , Líquido Cefalorraquidiano , Feminino , Seguimentos , Humanos , Aumento da Imagem , Pessoa de Meia-IdadeRESUMO
We report a 2-year-old girl with nephrotic syndrome, microcephaly, seizures and psychomotor retardation. Histological studies of a renal biopsy revealed focal glomerular sclerosis with mesangiolysis and capillary microaneurysms. Dysmorphic features were remarkable: abnormal-shaped skull, coarse hair, narrow forehead, large low-set ears, almond-shaped eyes, low nasal bridge, pinched nose, thin lips and micrognathia. Cases with this rare combination of microcephaly and early onset of nephrotic syndrome with various neurological abnormalities have been reported. However, clinical manifestations and histological findings showed a wide variation, and there is a lot of confusion in this syndrome. We therefore reviewed the previous reports and propose a new classification of this syndrome.
Assuntos
Deficiência Intelectual/diagnóstico , Microcefalia/diagnóstico , Síndrome Nefrótica/diagnóstico , Convulsões/diagnóstico , Pré-Escolar , Diagnóstico Diferencial , Face/anormalidades , Feminino , Glomerulosclerose Segmentar e Focal/diagnóstico , Glomerulosclerose Segmentar e Focal/patologia , Humanos , Rim/patologia , Síndrome Nefrótica/patologia , Linhagem , SíndromeAssuntos
Ensaio de Imunoadsorção Enzimática/métodos , Glutationa Peroxidase/sangue , Imunoglobulinas/imunologia , Falência Renal Crônica/enzimologia , Diálise Renal , Adulto , Animais , Anticorpos/imunologia , Especificidade de Anticorpos , Galinhas , Nefropatias Diabéticas/enzimologia , Gema de Ovo/imunologia , Feminino , Glomerulonefrite/enzimologia , Glutationa Peroxidase/imunologia , Humanos , Falência Renal Crônica/sangue , Masculino , Pessoa de Meia-Idade , Doenças Renais Policísticas/enzimologia , Gravidez , Complicações na Gravidez/enzimologia , Reprodutibilidade dos TestesAssuntos
Síndrome de Bartter/terapia , Hormônio do Crescimento/uso terapêutico , Estatura , Criança , Humanos , Masculino , SíndromeAssuntos
Glomerulonefrite/etiologia , Hepatite B/complicações , Animais , Glomerulonefrite/imunologia , Glomerulonefrite Membranoproliferativa/etiologia , Glomerulonefrite Membranosa/etiologia , Hepatite B/genética , Hepatite B/imunologia , Antígenos E da Hepatite B/imunologia , Humanos , Doenças do Complexo Imune/etiologiaRESUMO
To evaluate renal terminal complement activation in patients with glomerular diseases, we measured terminal complement complexes (TCCs) in plasma and urine with sandwich enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody against a C9 neoepitope expressed on TCC and a polyclonal antihuman C7 antibody. TCCs were detectable in plasma but not in urine in most of normal controls. In plasma, TCC levels were elevated in 4 of 22 patients with lupus nephritis and in 6 of 12 with membranoproliferative glomerulonephritis. However all patients with IgA nephritis, focal glomerulosclerosis, idiopathic membranous nephritis and idiopathic minimal change nephrotic syndrome (MC) showed normal values. In urine, TCCs were detectable in almost all patients with heavy proteinuria (greater than or equal to 100 mg/ml) except MC. The TCCs present in urine were partially purified by gel filtration using Sepharose 6B and were found to contain C5, C6, C7, C8, C9 and S protein by ELISA. Although the molecular weight of TCC is similar to that of IgM, the fractional excretion rate of TCC was about 100 times higher than that of IgM. These results suggest that TCCs detectable in urine contain SC5b-9 complexes and are mostly of renal origin.
Assuntos
Complexo de Ataque à Membrana do Sistema Complemento/urina , Nefropatias/imunologia , Adolescente , Adulto , Criança , Feminino , Glomerulonefrite por IGA/imunologia , Glomerulonefrite Membranoproliferativa/imunologia , Glomerulonefrite Membranosa/imunologia , Glomerulosclerose Segmentar e Focal/imunologia , Humanos , Glomérulos Renais/imunologia , Nefrite Lúpica/imunologia , Masculino , Pessoa de Meia-IdadeRESUMO
We describe a 13-year-old girl with the incomplete type of Behçet's disease who had recurrent oral and genital ulcers, folliculitis, proteinuria and hematuria. Renal biopsy specimens revealed diffuse proliferative glomerulonephritis with strongly positive IgA deposits in the glomerular mesangial area, which is histologically indistinguishable from primary IgA nephritis. Further studies of the IgA subclasses showed that IgA1 deposits were predominant in the glomerular mesangium. Primary IgA nephritis is thought to be associated with polymeric IgA1. So it appears that there may be a common underlying disease or mechanism involved in both primary IgA nephritis and the IgA nephritis in Behçet's disease.
Assuntos
Síndrome de Behçet/complicações , Glomerulonefrite por IGA/etiologia , Adolescente , Adulto , Síndrome de Behçet/imunologia , Síndrome de Behçet/patologia , Criança , Feminino , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
A 10-year-old boy had short stature, external ophthalmoplegia, atypical retinal pigmentary degeneration, and sensorineural hearing loss (Kearns-Sayre syndrome). In addition to ragged-red fibers observed on modified Gomori trichrome staining, there were scattered fibers exhibiting no cytochrome c oxidase activity, indicating a focal deficiency. Cytochrome c oxidase and other respiratory chain enzyme activities were normal biochemically. The patient also had renal tubular dysfunction, including isosthenuria, decreased urine-concentrating ability, and excessive excretion of potassium and magnesium. In addition, he had hyperreninemia and hyperaldosteronism but no hypertension. The renal dysfunction was thought to have resulted from a primary defect in the thick ascending limb of the loop of Henle, mimicking Bartter syndrome. In contrast to previously described cases of cytochrome c oxidase deficiency with de Toni-Fanconi Debré syndrome, the patient had less intensive muscle abnormalities. A renal biopsy specimen showed ultrastructural changes in mitochondria that were similar to those seen in biopsy specimens of muscle. A large-scale deletion (8.8 kilobases) in mitochondrial DNA was found in biopsy specimens of muscle and kidney.