RESUMO
The (pro)renin receptor ((P)RR) is cleaved to generate soluble (P)RR (s(P)RR), which reflects the status of the tissue renin-angiotensin system. Hemodialysis (HD) patients have a poor prognosis due to the increased prevalence of cardiovascular diseases. The present study aimed to investigate whether serum s(P)RR level is associated with the worsening of cardiac function in HD patients. A total of 258 maintenance HD patients were recruited and serum s(P)RR concentration was measured. Background factors in patients who survived (S group) and patients who died (D group) during the 12-month follow-up period and relationships between serum s(P)RR level and changes in cardiac function during the follow-up period in the S group were investigated. The median serum s(P)RR value at baseline was 29.8 ng/ml. Twenty-four patients died during the follow-up period. Cardiothoracic ratio, human atrial natriuretic peptide (hANP), brain natriuretic peptide (BNP), and E over e-prime were significantly higher in the D group. In the S group, changes in hANP or BNP were significantly greater in the higher serum s(P)RR group than in the lower serum s(P)RR group. High serum s(P)RR level was significantly correlated with changes in BNP, independent of other factors. High serum s(P)RR level was associated with increases in BNP, independent of other risk factors, suggesting that an increased expression of (P)RR may be associated with a progression of heart failure in HD patients and that serum s(P)RR concentration could be used as a biomarker for selecting patients requiring intensive care.
Assuntos
Biomarcadores/sangue , Doenças Cardiovasculares/diagnóstico , Doenças Cardiovasculares/mortalidade , Receptores de Superfície Celular/sangue , Diálise Renal/efeitos adversos , ATPases Vacuolares Próton-Translocadoras/sangue , Idoso , Doenças Cardiovasculares/sangue , Doenças Cardiovasculares/etiologia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Taxa de SobrevidaRESUMO
Each year, many fatalities result from rollovers of agricultural tractors in Japan. In addition to rollover protective structures (ROPS) and seat belts, a warning device that alerts the operator of impending rollover based on the tractor stability index is a measure used to prevent rollovers. The stability index requires inertial parameters, which have been measured only for the single body of the tractor, to calculate the warning threshold. In this study, the center of gravity (CoG) and lateral stability angles of three agricultural tractors were measured, and lateral stability angles were also calculated and compared with measured values for three tractor-tiller combinations to analyze the effect of the attached implement on the tractor stability as well as to verify the accuracy of the calculation methods. The roll moment of inertia (RMI) was also measured for two tractors and two rotary tillers, and RMI values for tractor-tiller combinations were calculated. The measurement and calculation results show that the attached implement increased the lateral stability angle of tractors in phase I rollover and decreased the lateral stability angle in phase II rollover, and for a certain tractor-tiller combination, there was no transition from phase I to phase II rollover. The difference between the measured and calculated lateral stability angles in phase I ranged from -3.5° to 2.5°, while that in phase II ranged from 0.2° to 5.2°. The RMI about the longitudinal axis through the CoG was 203 and 433 kg m-2 for tractors A and B, respectively, and 52 and 94 kg m-2 for rotary tillers D and F, respectively. The calculated RMI values were 265 and 540 kg m-2 for tractor-tiller combinations A-D and B-E, respectively.
Assuntos
Acidentes de Trabalho/prevenção & controle , Agricultura/instrumentação , Segurança de EquipamentosRESUMO
Highly organized cell processes characterize glomerular podocytes in vivo. However, podocytes in culture have a simple morphology lacking cell processes, especially upon reaching confluence. Here, we aimed to establish culture conditions under which cultured podocytes extend cell processes at confluence. Among various culture conditions that could possibly cause phenotypic changes in podocytes, we examined the effects of heparin, all-trans retinoic acid, fetal bovine serum, and extracellular matrices on the morphology of podocytes in rat primary culture. Consequently, long arborized cell processes were observed to radiate extensively from the cell body only when cells were cultured in the presence of heparin and all-trans retinoic acid on laminin-coated dishes with decreasing concentrations of fetal bovine serum. Primary processes branching repeatedly into terminal processes and cell process insertion under adjacent cell bodies were evident by electron microscopy-based analysis. Immunostaining for podocin showed conspicuous elongations of intercellular junctions. Under these conditions, the expression levels of podocyte-specific proteins and genes were markedly upregulated. Thus, we succeeded in establishing culture conditions in which the cultured podocytes exhibit phenotypes similar to those under in vivo conditions.
Assuntos
Técnicas de Cultura de Células , Forma Celular , Extensões da Superfície Celular/ultraestrutura , Podócitos/ultraestrutura , Animais , Proliferação de Células , Forma Celular/efeitos dos fármacos , Extensões da Superfície Celular/efeitos dos fármacos , Células Cultivadas , Regulação da Expressão Gênica , Heparina/farmacologia , Laminina/metabolismo , Masculino , Fenótipo , Podócitos/efeitos dos fármacos , Podócitos/metabolismo , Ratos Wistar , Fatores de Tempo , Tretinoína/farmacologiaRESUMO
BACKGROUND: A dacryolith mainly contains organic material, but its specific protein content is unknown. We observed a unique dacryolith formation in patients with long-term use of rebamipide eye drops and tried to identify the differences in protein compositions of dacryoliths from patients with or without use of rebamipide eye drops using novel proteomic analysis. METHODS: Dacryolith samples were obtained from 7 patients (4 samples were without rebamipide usage, 3 were with rebamipide usage) who underwent endo-dacryocystorhinostomy or lacrimal endoscopic surgery and were subjected to protein identification and metaproteomic analysis. RESULTS: The proteomic analysis revealed that most core proteins of dacryoliths are involved in immune and inflammatory responses and rebamipide-related proteins participated in several biological processes, including immune response, receptor-mediated endocytosis, and negative regulation of endopeptidase activity. Metaproteomic analysis of taxonomic diversity of dacryolith proteomes suggested less involvement of bacterial infections in dacryoliths from patients with long-term use of rebamipide. CONCLUSION: This is the first report to clinically detect the different features of dacryoliths related with long-term use of rebamipide eye drops with proteomic analysis. It is possible that lacrimal protein alteration may be accelerated by topical rebamipide treatment with less bacterial infection involvement, but this requires further study. Long-term rebamipide eye drop use may be restricted in patients with nasolacrimal duct obstruction.
RESUMO
Nephrin, a major intercellular junction (ICJ) molecule of mammalian podocytes in the renal glomerulus, is absent in the avian genome. We hypothesized that birds use ICJ molecules other than nephrin in their podocytes. Therefore, in the present study, we examined the possible involvement of adherens junction (AJ) proteins in the ICJs of avian podocytes. We found the AJ proteins N-cadherin and α- and ß-catenins in podocytes of quail and chickens but not in those of rats, pigs or humans. The AJ proteins were prominent in avian glomerulus-rich fractions in immunoblot analyses, and in immunofluorescence microscopy analyses, they were localized along glomerular capillary walls appearing in at least two staining patterns: weakly diffuse and distinctly granular. Immunoelectron microscopy demonstrated that the significant accumulation of immunogold particles for the AJ proteins were especially evident in avian slit diaphragms and AJs. Furthermore, N-cadherin was found to be expressed in all nephron cells in the early developmental stage but became confined to podocytes during maturation. These results indicate that avian slit diaphragms clearly express AJ proteins as compared with that in the mammal-where AJ proteins are suppressed to an extremely low level-and that avian podocytes are interconnected by AJs per se in addition to slit diaphragms.
Assuntos
Junções Aderentes/química , Junções Aderentes/metabolismo , Proteínas de Membrana/deficiência , Podócitos/química , Podócitos/metabolismo , Animais , Caderinas/metabolismo , Cateninas/metabolismo , Galinhas , Coturnix , Feminino , Humanos , Ratos , Ratos Wistar , SuínosRESUMO
OFFGEL fractionation of mouse kidney protein lysate and its tryptic peptide digest has been examined in this study for better understanding the differences between protein and peptide fractionation methods and attaining maximum recruitment of this modern methodology for in-depth proteomic analysis. With the same initial protein/peptide load for both fractionation methods, protein OFFGEL fractionation showed a preponderance in terms of protein identification, fractionation efficiency, and focusing resolution, while peptide OFFGEL was better in recovery, number of peptide matches, and protein coverage. This result suggests that the protein fractionation method is more suitable for shotgun analysis while peptide fractionation suits well quantitative peptide analysis [isobaric tags for relative and absolute quantitation (iTRAQ) or tandem mass tags (TMT)]. Taken together, utilization of the advantages of both fractionation approaches could be attained by coupling both methods to be applied on complex biological tissue. A typical result is shown in this article by identification of 8262 confident proteins of whole mouse kidney under stringent condition. We therefore consider OFFGEL fractionation as an effective and efficient addition to both label-free and quantitative label proteomics workflow.
Assuntos
Peptídeos/análise , Proteínas/análise , Proteômica , Animais , Cromatografia Líquida de Alta Pressão , Cromatografia de Fase Reversa , Eletroforese em Gel de Poliacrilamida , Focalização Isoelétrica , Rim/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Peptídeos/isolamento & purificação , Peptídeos/metabolismo , Proteínas/isolamento & purificação , Proteínas/metabolismo , Quinolinas/química , Espectrometria de Massas em Tandem , Tripsina/metabolismoRESUMO
A 21-year-old woman with nephrotic syndrome was referred to our hospital. She had congenital diaphragmatic hernia, hypoxic ischemic encephalopathy, and mental retardation, and had been treated for hyperthyroidism with thiamazole in another hospital. Serum creatinine was 37.8 µmol/L and antineutrophil cytoplasmic antibody against myeloperoxidase (MPO-ANCA) was 39 EU. Urinalyses were 3+ for proteins and 3+ for occult blood. A renal biopsy was performed. An examination using light microscopy (LM) revealed necrotizing glomerulonephritis with crescent formation. Immunofluorescence microscopy showed granular staining with immunoglobulin G and complement component 3 along the capillary walls. Electron microscopy (EM) disclosed subepithelial dense deposits. A renal biopsy suggested necrotizing glomerulonephritis with membranous nephropathy (MN) in stages I or II. Since many cases of drug-induced ANCA-associated glomerulonephritis (AAG) have been reported, we stopped thiamazole and treated with corticosteroid. The MPO-ANCA titer became negative 49 days after the initiation of treatment. Two years after the first treatment, the MPO-ANCA titer became elevated again and was 82 EU. The patient was administered cyclophosphamide and prednisone. However, the MPO-ANCA titer did not decrease. A renal biopsy was performed again 3 years after the first renal biopsy. LM revealed no crescentic formation but demonstrated spike formations along the glomerular basement membrane. EM also disclosed subepithelial dense deposits, but less than the first biopsy. The renal biopsy suggested MN in stages II or III. AAG was regarded as inactive after corticosteroid treatment. Therefore, ciclosporin administration was started. In conclusion, we experienced a rare case of AAG complicated with MN. The histopathologic results showed that immunosuppressive therapy seemed to be effective in treating crescentic glomerulonephritis; furthermore, it reduced proteinuria but could not reduce the MPO-ANCA titer.