RESUMO
AIMS/HYPOTHESIS: The NPHS1 gene product, nephrin, is a crucial component of the glomerular filtration barrier preventing proteinuria and previously assumed to be kidney-specific. The aim of this study was to describe the expression of nephrin mRNA and protein in human pancreas as well as identify the nephrin-expressing cell types. METHODS: RNA dot blot, reverse transcriptase-polymerase chain reaction, sequencing, immunoblotting and dual immunofluorescence were used for the characterisation of nephrin in the pancreas. RESULTS: Except for the kidney, the pancreas was found to be the only tissue expressing nephrin as screened with a human tissue RNA dot blot. The expression was verified with reverse transcriptase-polymerase chain reaction and by sequencing nephrin from a human pancreatic complementary DNA library. Nephrin antibody in immunoblot detected a 165,000 M(r) protein in the pancreas. Dual immunofluorescence showed that nephrin was specifically localised in the beta cells of the islets of Langerhans. There was no overlap with glucagon, somatostatin, or the ductal cell marker cytokeratin 19. CONCLUSION/INTERPRETATION: These data show that nephrin is a novel molecule of pancreatic beta cells.
Assuntos
Expressão Gênica , Ilhotas Pancreáticas/metabolismo , Proteínas/genética , Cadáver , Fluoresceína-5-Isotiocianato , Imunofluorescência , Corantes Fluorescentes , Glucagon/análise , Humanos , Immunoblotting , Queratinas/análise , Rim/química , Rim/embriologia , Córtex Renal/química , Proteínas de Membrana , Peso Molecular , Pâncreas/química , Proteínas/análise , RNA Mensageiro/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Somatostatina/análiseRESUMO
Diabetic nephropathy is a major complication of diabetes leading to thickening of the glomerular basement membrane, glomerular hypertrophy, mesangial expansion, and overt renal disease. The pathophysiologic mechanisms of diabetic nephropathy remain poorly understood. Nephrin is a recently found podocyte protein crucial for the interpodocyte slit membrane structure and maintenance of an intact filtration barrier. Here we have assessed the role of nephrin in two widely used animal models of diabetes, the streptozotocin model of the rat and the nonobese diabetic mouse. In both models, the expression levels of nephrin-specific mRNA as determined by real-time quantitative polymerase chain reaction increased up to two-fold during several weeks of follow-up. Immunohistochemical stainings revealed nephrin also more centrally within the glomerular tuft along with its preferential site in podocytes. Interestingly, as detected by immunoblotting, nephrin protein was also found in the urine of streptozotocin-induced rats. We conclude that nephrin is connected to the early changes of diabetic nephropathy and thus may contribute to the loss of glomerular filtration function.
Assuntos
Nefropatias Diabéticas/genética , Expressão Gênica , Proteínas/genética , Animais , Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/urina , Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/urina , Glomérulos Renais/metabolismo , Masculino , Proteínas de Membrana , Proteínas/metabolismo , RNA Mensageiro/sangue , RNA Mensageiro/urina , Ratos , Ratos Wistar , Distribuição TecidualRESUMO
Baccalaureate and graduate nursing education programs now have a choice of professional accrediting bodies and must make a decision about which best suits the needs of their individual programs. The authors describe the accreditation process and procedures of the National League for Nursing Accrediting Commission and the Commission on Collegiate Nursing Education. The purpose, philosophy, standards, accreditation process, fees, review cycle, and length of accreditation are compared.
Assuntos
Acreditação/normas , Bacharelado em Enfermagem , Educação de Pós-Graduação em Enfermagem , Acreditação/economia , Honorários e Preços , Humanos , Objetivos Organizacionais , Estados UnidosRESUMO
Nephrin is a novel transmembrane protein of kidney glomerular podocytes, which appears crucially important for the maintenance of the glomerular filtration barrier. According to its predicted structure, nephrin has additional roles in cell-cell adhesion and/or signal transduction. We have previously cloned the rat homologue of nephrin and described its alternatively spliced transcripts alpha and beta. In this study we examined the alterations in expression and regulation of particularly the major alternatively spliced nephrin-alpha giving rise to a variant lacking the membrane spanning domain in the puromycin nephrosis of the rat. A down-regulation of up to 78% was observed of the full length mRNA after 10 d of PAN treatment. The expression changes of nephrin-alpha followed closely the expression of the full length mRNA. Interestingly, we also found nephrin protein in urine at the peak proteinuria samples of this model. These results suggest that soluble nephrin variants may be important markers for proteinuric diseases.
Assuntos
Processamento Alternativo , Síndrome Nefrótica/genética , Proteínas/genética , RNA Mensageiro/metabolismo , Animais , Western Blotting , Glomérulos Renais/metabolismo , Masculino , Proteínas de Membrana , Síndrome Nefrótica/induzido quimicamente , Síndrome Nefrótica/complicações , Estrutura Terciária de Proteína , Proteinúria/etiologia , Puromicina/toxicidade , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Relação Estrutura-AtividadeRESUMO
BACKGROUND: The recently identified gene NPHS1 with its mutations causing congenital nephrotic syndrome of the Finnish type (CNF) is highly promising in providing new understanding of pathophysiology of proteinuria. Earlier we cloned a rat NPHS1 homologue, as well as characterized and raised antibodies to the respective protein product nephrin. METHODS: Changes in the expression levels of nephrin-specific mRNA in commonly used experimental models of proteinuria were examined using semiquantitative reverse transcription-polymerase chain reaction, immunofluorescence, and immunoelectron microscopy (IEM) of nephrin. RESULTS: Notably, a 40% down-regulation of the nephrin-specific mRNA of cortical kidney was seen already at day 3 after induction of the puromycin aminonucleoside nephrosis (PAN), while no major elevation of urinary protein secretion was seen at this stage. A further decrease of 80% of nephrin message was seen at the peak of proteinuria at day 10. A similar decrease of up to 70% from the basal levels was seen in mercuric chloride-treated rats. Changes in the protein expression paralleled those of the mRNA in indirect immunofluorescence. Interestingly, a remarkable plasmalemmal dislocation from the normal expression site at the interpodocyte filtration slits could be observed in IEM. CONCLUSIONS: Nephrin appears to be an important causative molecule of proteinuria and shows a remarkable redistribution from the filtration slits to the podocyte plasma membrane, especially in PAN.
Assuntos
Glomerulonefrite/metabolismo , Glomerulonefrite/fisiopatologia , Proteínas/genética , Proteínas/metabolismo , Albuminúria/induzido quimicamente , Albuminúria/metabolismo , Albuminúria/fisiopatologia , Animais , Antimetabólitos Antineoplásicos , Modelos Animais de Doenças , Desinfetantes , Expressão Gênica/fisiologia , Glomerulonefrite/induzido quimicamente , Glomérulos Renais/química , Glomérulos Renais/metabolismo , Glomérulos Renais/ultraestrutura , Peroxidação de Lipídeos/fisiologia , Masculino , Proteínas de Membrana , Cloreto de Mercúrio , Microscopia Imunoeletrônica , Proteínas/análise , Puromicina Aminonucleosídeo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transcrição Gênica/fisiologiaRESUMO
Nursing faculty and students, as well as schools of nursing, are concerned about maintaining or improving pass rates on the National Council Licensure Examination for Registered Nurses (NCLEX-RN). This article offers a comparative review of eight computerized NCLEX-RN preparation programs: Compass RN; Health Education Systems, Inc. (HESI) Assess Test; NLN-CAT-RN Pharmacology Test; Meds, incorporated Comprehensive Q & A; Mosby's NCLEX Review; Medi-Sim Computer-Assisted Preparation for NCLEX-RN; Preparing for NCLEX-RN CAT; and NLN-CAT Practice RN. Criteria for evaluating these programs include program design features, types of questions and feedback, performance reports, screen design, ease of use, technical support and documentation, warranty and upgrade policy, and price. Issues associated with using NCLEX-RN preparation programs also are discussed.