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Artigo Fluctuating jaundice in the adenocarcinoma of the ampulla of Vater: a classic sign or an exception?(FapUNIFESP (SciELO), 2015) Souza, Dyego Leandro Bezerra de; Alves, José Roberto; Amico, Enio Campos; Oliveira, Patrick Vanttinny Vieira de; Maranhão, Icaro Godeiro de Oliveira; https://orcid.org/0000-0001-8426-3120Background - Some authors consider the fluctuating jaundice as a classic sign of the adenocarcinoma of the ampulla of Vater. Objetive - Assessing the frequency of fluctuating jaundice in their forms of its depiction in the patients with adenocarcinoma of the ampulla of Vater. Methods - Observational and retrospective study, conducted through analyses of medical records from patients subjected to pancreatic cephalic resections between February 2008 and July 2013. The pathological examination of the surgical specimen was positive to adenocarcinoma of the ampulla of Vater. Concepts and differences on clinical and laboratory fluctuating jaundice were standardized. It was subdivided into type A and type B laboratory fluctuating jaundice. Results - Twenty patients were selected. One of them always remained anicteric, 11 patients developed progressive jaundice, 2 of them developed clinical and laboratory fluctuating jaundice, 5 presented only laboratory fluctuating jaundice and one did not present significant variations on total serum bilirubin levels. Among the seven patients with fluctuating jaundice, two were classified as type A, one as type B and four were not classified due to lack information. Finally, progressive jaundice was the prevailing presentation form in these patients (11 cases). Conclusion - This series of cases suggested that clinical fluctuating jaundice is a uncommon signal in adenocarcinoma of the ampulla of VaterArtigo Microbiological analysis of bile in patients with benign and malignant biliopancreatic diseases and its consequences(FapUNIFESP (SciELO), 2016) Souza, Dyego Leandro Bezerra de; Alves, José Roberto; Silva, Rodrigo do Carmo; Guerra, Sâmea Costa Pinheiro; Freitas, Tiago Tavares de; Amico, Enio Campos; https://orcid.org/0000-0001-8426-3120- Background - Bactibilia has several consequences to human health. Objetive - Assessing the bile microbiology of patients with biliopancreatic diseases in order to identify bacteria and their possible infectious complications. Methods - Retrospective study of 30 bile culture samples from patients with benign and malignant biliopancreatic diseases. The samples were assessed to set the bile microbiological flora and to search for its possible link with comorbidity, carcinogenesis and postoperative infectious complications. Results - Thirty bile samples from patients at mean age ≈57.7 years, mostly female (n=18), were assessed. Bactibilia was found in 12 cases, mostly in patients with benign diseases (n=8), older than 50 years (n=23) and female (n=10). Adenocarcinoma of the duodenal papilla (n=9) and cholelithiasis (n=8) were the most common diseases. Escherichia coli (n=5) and Klebsiella sp (n=3) were predominantly found in patients with benign diseases; and Klebsiella sp (n=2) and Streptococcus sp (n=2) were prevalent in cancer patients. There were postoperative infectious complications in seven cases, five of them in bactibilia-associated patients (P=0.084). Conclusion - Bactibilia was found in 12 samples and Escherichia coli and Klebsiella sp were most often identified in patients with benign diseases, as well as Streptococcus sp and Klebsiella sp in cancer patients. There was a trend of higher postoperative infectious complication incidence in patients with bactibiliaArtigo Factors associated with mortality in hepatitis C patients(Revista Ciência Plural, 2017) Souza, Dyego Leandro Bezerra de; Sousa, Gilmar Amorim de; Pessoa, Ranna Santos; Souza Filho, Marlon César Melo; Oliveira, Daniel Fernandes Mello; Medeiros, Luana Lopes; Souza, Dyeo Leandro Bezerra; Costa, Iris do Céu Clara; https://orcid.org/0000-0001-8426-3120IIntrodução: A hepatite C continua sendo a maior causa de doença hepática progressiva que evolui para forma crônica em 80% dos pacientes agudamente infectados, podendo desencadear cirrose, hemorragia digestiva, falência hepática, câncer de fígado e morte. Objetivos: Conhecer fatores associados ao óbito em pacientes com hepatite C. Métodos:Revisou-se 10.304 prontuários, do Núcleo de Estudos do Fígado do Hospital Universitário Onofre Lopes da Universidade Federal do Rio Grande do Norte, Brasil, entre maio-1995 e maio-2013. Considerou-se casos suspeitos pacientes com anti-HCV positivo e casos confirmados, aqueles com HCV RNA qualitativo positivo, que resultou em 512 casos. O óbito foi a variável dependente. Foram consideradas variáveis independentes: as sócio-demográficas,as associadas à infecção pelo HCV e as relacionadas à progressão da doença. Avaliou-se associação das variáveis independentes e o óbito, e calculou-se significância estatística (p), Odds Ratio (OR) e intervalos de confiança de 95% (IC 95%). Resultados:Encontrou-se as seguintes associações com mortalidade por hepatite C: pacientes com idade acima de 35 anos, com abandono do tratamento, diabete melito, uso de insulina, bilirrubina total acima de 1,3 mg/dL, INR na consulta final e albumina baixa (<3,5g/dL) na consulta inicial, AST, ALT, TAP alargado, cirrose e hepatocarcinoma. Conclusão:Conclui-se que em função de sua magnitude e severidade, o fator de maior impacto na hepatite C é a descoberta precoce da doença, antes de evoluir para cirrose e carcinoma hepático, o que presume que esses pacientes precisam ter acesso facilitado ao serviço de saúde, que deverá ser garantido por políticas públicas específicas definidasArtigo Hypervascular liver lesions in radiologically normal liver(FapUNIFESP (SciELO), 2017) Souza, Dyego Leandro Bezerra de; Amico, Enio Campos; Alves, José Roberto; Salviano, Fellipe Alexandre Macena; João, Samir Assi; https://orcid.org/0000-0001-8426-3120Background: The hypervascular liver lesions represent a diagnostic challenge. Aim: To identify risk factors for cancer in patients with non-hemangiomatous hypervascular hepatic lesions in radiologically normal liver. Method: This prospective study included patients with hypervascular liver lesions in radiologically normal liver. The diagnosis was made by biopsy or was presumed on the basis of radiologic stability in follow-up period of one year. Cirrhosis or patients with typical imaging characteristics of haemangioma were excluded. Results: Eightyeight patients were included. The average age was 42.4. The lesions were unique and were between 2-5 cm in size in most cases. Liver biopsy was performed in approximately 1/3 of cases. The lesions were benign or most likely benign in 81.8%, while cancer was diagnosed in 12.5% of cases. Univariate analysis showed that age >45 years (p< 0.001), personal history of cancer (p=0.020), presence of >3 nodules (p=0.003) and elevated alkaline phosphatase (p=0.013) were significant risk factors for cancer. Conclusion: It is safe to observe hypervascular liver lesions in normal liver in patients up to 45 years, normal alanine aminotransaminase, up to three nodules and no personal history of cancer. Lesion biopsies are safe in patients with atypical lesions and define the treatment to be established for most of these patientsArtigo Anunusual case of bilateral macular detachment preceding renal failure(National Center for Biotechnology Information, 2021) Oliveira, Rodrigo Azevedo de; Garcia Filho, Carlos A. de Amorim; Meirelles, Rodrigo L.; Lima, Luiz H.; Balaratnasingam, Chandrakumar; Agarwal, Anita; Garcia, Carlos A. de AmorimA 58-year-old woman presented with a for 4-month history of prograssively decreasing visual acuity in both eyes and not other ophthalmic or systemic symptoms. Her medical history was relevant only for moderately controlled systemic hypertension.Artigo Confiabilidade do teste da caminhada de seis minutos em pacientes com miastenia gravis generalizada(Fisioterapia e Pesquisa, 2009) Dourado Junior, Mário Emílio Teixeira; Resqueti, Vanessa Regiane; Oliveira, Georges Willeneuwe de Sousa; Andrade, Armèle Dornelas de; Casan, Pere; Fregonezi, Guilherme Augusto de Freitas; https://orcid.org/0000-0002-9462-2294Este estudo objetivou determinar a confiabilidade do teste da caminhada de seis minutos (TC6M) como um teste de capacidade funcional em pacientes com miastenia gravis generalizada (MG). Foram selecionados 11 pacientes com MG - 5 homens, 6 mulheres - com idade de 55±9 anos, avaliados inicialmente quanto à função fulmonar, que se submeteram a três TC6M em dias diferentes. Durante e/ou após cada teste foram medidas freqüência cardíaca e saturação de oxigênio (por oxímetro portátil), sensação de dispnéia (pela escala de Borg) e distância percorrida. Nos três testes as distâncias percorridas foram 498 m, 517 m e 520 m (respectivamente 99%, 103% e 104% do valor predito). Em média, a freqüência cardíaca, dispnéia e saturação de oxigênio mostraram comportamento constante nos três testes. Foram encontradas alta confiabilidade relativa, com coeficiente de correlação interclasse maior que 0,90 entre os testes (TC6M1-TC6M2, 0,960; TC6M1-TC6M3, 0,945; e TC6M2-TC6M3, 0,970) e confiabilidade absoluta de 4%, 3,5% e 4,8%, com reprodutibilidade de 11%, 9,8% e 13,4%, respectivamente para o primeiro, segundo e terceiro testes. Os limites superiores e inferiores de concordância e o valor médio das médias das diferenças (bias) calculados pelo teste de Bland-Altman mostraram-se clinicamente aceitáveis. Conclui-se que o TC6M se mostrou seguro, confiável e reprodutível, podendo ser aplicado para avaliação e seguimento da tolerância ao exercício em pacientes com MG generalizada.Artigo A heterotrimeric GProtein, G«i-3 , on Golgi membranes regulates the secretion of a heparan sulfate proteoglycan in LLC-PKI epithelial cells(The Journal Of Cell Biology, 1991) Almeida, Jose Bruno de; Stow, Jennifer L.; Narula, Navneet; Holtzman, Eliezer J.; Ercolani, Louis; Ausiello, Dennis A.A heterotrimeric Gai subunit, «i- 3 , is localized on Golgi membranes in LLC-PKI and NRK epithelial cells where it colocalizes with mannosidase II by immunofluorescence . The M-3 was found to be localized on the cytoplasmic face of Golgi cisternae and it was distributed across the whole Golgi stack. The a;_3 subunit is found on isolated rat liver Golgi membranes by Western blotting and Gai -3 on the Golgi apparatus is ADP ribosylated by pertussis toxin. LLCPKI cells were stably transfected with GO;-3 on an MT1, inducible promoter in order to overexpress ai-3 on Golgi membranes. The intracellular processing and constitutive secretion of the basement membrane heparan sulfate proteoglycan (HSPG) was measured in LLC-PK, cells. Overexpression of ai-3 on Golgi membranes in transfected cells retarded the secretion of HSPG and accumulated precursors in the medial-trans-Golgi . This effect was reversed by treatment of cells with pertussis toxin which results in ADP-ribosylation and functional uncoupling of Gai-3 on Golgi membranes. These results provide evidence for a novel role for the pertussis toxin sensitive Gai-3 protein in Golgi trafficking of a constitutively secreted protein in epithelial cellsArtigo Purified epithelial Na+ channel complex contains the pertussis toxin-sensitive Gai-3 protein(Elsevier, 1992) Almeida, José Bruno de; Ausiello, Dennis A.; Stow, Jennifer L.; Cantiello, Horacio F.; Benos, Dale J.We have recently demonstrated that the amiloride-sensitive Na+ channel in the apical membrane of the renal epithelial cell line, A6, is modulated by the alpha i-3 subunit of the Gi-3 protein. We also showed that a 700-kDa protein complex can be purified from the membranes of A6 epithelia which (a) can reconstitute the amiloride-sensitive Na+ influx in liposomes and planar bilayer membranes and (b) consists of six major protein bands observed on reducing sodium dodecyl sulfate-polyacrylamide gels with molecular masses ranging from 35 to 320 kDa. The present study was undertaken to determine if the alpha i-3 subunit was a member of this Na+ channel complex. G alpha i structure and function were identified by Western blotting with specific G alpha i subunit antibodies and Na+ channel antibodies, through ADP-ribosylation with pertussis toxin, and by immunocytochemical localization of the Na+ channel and G alpha i proteins. We demonstrate that two protein substrates are ADP-ribosylated in the 700-kDa complex in the presence of pertussis toxin and are specifically immunoprecipitated with an anti-Na+ channel polyclonal antibody. One of these substrates, a 41-kDa protein, was identified as the alpha i-3 subunit of the Gi-3 protein on Western blots with specific antibodies. Na+ channel antibodies do not recognize G alpha i-3 on Western blots of Golgi membranes which contain alpha i-3 but not Na+ channel proteins, nor do they immunoprecipitate alpha i-3 from solubilized Golgi membranes; however, alpha i-3 is coprecipitated as part of the Na+ channel complex from A6 cell membranes by polyclonal Na+ channel antibodies. Both alpha i-3 and the Na+ channel have been localized in A6 cells by confocal imaging and immunofluorescence with specific antibodies and are found to be in distinct but adjacent domains of the apical cell surface. In functional studies, alpha i-3, but not alpha i-2, stimulates Na+ channel activity. These data are therefore consistent with the localization of Na+ channel activity and modulatory alpha i-3 protein at the apical plasma membrane, which together represent a specific signal transduction pathway for ion channel regulationArtigo Severe hypertension induces disturbances of renal autoregulation(Hypertension, 1992) Almeida, Jose Bruno de; Saragoca, Manoel A.; Tavares, Agostinho; Cezareti, Mário L.; Draibe, Sérgio A.; Ramos, Oswaldo L.To study if the severity of hypertension could be associated with disturbances of the autoregulation of renal blood flow and glomerular filtration, we compared the renal hemodynamic and functional responses to acute blood pressure reductions of a group of patients with moderate essential hypertension (n = 10) with those of a group of patients with severe hypertension (n = 10). Blood pressure was reduced to normal levels by a stepwise infusion of sodium nitroprusside, and effective renal blood flow (by 131I-hippuran), glomerular filtration rate (by endogenous creatinine clearance), and filtration fraction were determined. After acute blood pressure normalization, effective renal blood flow and glomerular filtration rate were significantly reduced in patients with severe hypertension (-41.6 +/- 8.3% and -44.7 +/- 6.8%, respectively; p less than 0.01 for both) but not in those with moderate hypertension (+4.9 +/- 9.1% and +6.2 +/- 13.3%, respectively; NS). Filtration fraction remained unchanged in both groups. These results show that severe but not moderate essential hypertensive patients have a displacement to the right of the lower limit of the renal autoregulation curve due to impaired vasodilation to maintain adequate renal blood flow during acute reductions of blood pressure. This impairment may be due to anatomic or functional defects of preglomerular vessels, or to both. Furthermore, the inability to maintain adequate glomerular filtration in these circumstances shows that patients with severe hypertension also have an impaired ability to adjust postglomerular vasomotor tone in the face of reductions in glomerular blood flow.Artigo Binding of the cytosolic p200 protein to Golgi membranes is regulated by heterotrimeric G proteins(Journal Of Cell Science, 1993) Almeida, Jose Bruno de; Doherty, Joanne; Ausiello, Dennis A.; Stow, Jennifer L.The formation of vesicles for protein trafficking requires the dynamic binding of cytosolic coat proteins onto Golgi membranes and this binding is regulated by a variety of GTPases, including heterotrimeric G proteins. We have previously shown the presence of the pertussis toxin-sensitive G i-3 protein on Golgi membranes and demonstrated a functional role for G i-3 in the trafficking of secretory proteins through the Golgi complex. We have also described a brefeldin A-sensitive phosphoprotein, p200, which is found in the cytoplasm and on Golgi membranes. The present study investigates the role of heterotrimeric G proteins in the regulation of p200 binding to Golgi membranes. An in vitro binding assay was used to measure the binding of cytosolic p200 to LLC-PK1 cell microsomal membranes and to purified rat liver Golgi membranes in the presence of specific activators of G proteins. The binding of p200 to Golgi membranes was compared to that of the coatomer protein -COP, for which G protein-dependent membrane binding has previously been established. Membrane binding of both p200 and -COP was induced maximally by activation of all G proteins in the presence of GTP S. More selective activation of the heterotrimeric G proteins, with AlFn or mastoparan, also induced membrane binding of p200 and -COP. Pertussis toxin pretreatment of Golgi membranes, to selectively inactivate G i-3, reduced the AlFn and mastoparan-induced binding of p200 to Golgi membranes, whereas no significant effect of pertussis toxin on -COP binding was found in this assay. The effect of pertussis toxin thus implicates G i-3, as one component of a regulatory pathway, in the binding of cytosolic p200 to Golgi membranes. The effects of AlFn and pertussis toxin on p200 membrane binding were also shown in intact cells by immunofluorescence staining. AlFn treatment of cells induced translocation of p200 from the cytoplasm onto the Golgi complex, resulting in a conformational change in some Golgi membranes. The translocation of p200 was blocked by pretreatment of intact NRK cells with pertussis toxin. The data presented here support the conclusion that the binding of the p200 protein to Golgi membranes involves regulation by the pertussis toxinsensitive heterotrimeric G proteins, specifically the G i-3 proteinArtigo Entry of cholera toxin into polarized human intestinal epithelial cells. Identification of an early brefeldin a sensitive event required for A1-peptide generation(American Society for Clinical Investigation, 1993) Almeida, Jose Bruno de; Lencer, W. I.; Moe, S.; Stow, Jennifer L.; Ausiello, Dennis A.; Madara, J. L.The effect of brefeldin-A (BFA), a reversible inhibitor of vesicular transport, on cholera toxin (CT)-induced Cl- secretion (Isc) was examined in the polarized human intestinal cell line, T84. Pretreatment of T84 monolayers with 5 microM BFA reversibly inhibited Isc in response to apical or basolateral addition of 120 nM CT (2.4 +/- 0.5 vs. 68 +/- 3 microA/cm2, n = 5). In contrast, BFA did not inhibit Isc responses to the cAMP agonist VIP (63 +/- 7 microA/cm2). BFA had no effect on cell surface binding and endocytosis of a functional fluorescent CT analog or on the dose dependency of CT induced 32P-NAD ribosylation of Gs alpha in vitro. In contrast, BFA completely inhibited (> 95%) the ability of T84 cells to reduce CT to the enzymatically active A1-peptide. BFA had to be added within the first 10 min of CT exposure to inhibit CT-elicited Isc. The early BFA-sensitive step occurred before a temperature-sensitive step essential for apical CT action. These studies show that sequential steps are required for a biological response to apical CT: (a) binding to cell surfaces and rapid endocytosis; (b) early, BFA-sensitive vesicular transport essential for reduction of the A1-peptide; and (c) subsequent temperature-sensitive translocation of a signal (the A1-peptide or possibly ADP-ribose-Gs alpha) to the basolateral domainArtigo Distribution and role of heterotrimeric G proteins in the secretory pathway of polarized epithelial cells(Journal Of Cell Science, 1993) Almeida, Jose Bruno de; Stow, Jennifer L.The movement of newly synthesized proteins in the constitutive secretory pathway, from their site of synthesis in the endoplasmic reticulum to the cell surface or to intracellular destinations, requires an orderly sequence of transport steps between membrane-bound compartments. Until recently, the trafficking and secretion of proteins through this pathway was thought to occur as a relatively automatic, unregulated series of events. Recent studies show that protein trafficking in the constitutive secretory pathway requires G T P hydrolysis by families of GTP-binding proteins (G proteins), which at multiple steps potentially provide regulation and specificity for protein trafficking. M any monomeric G proteins are known to be localized and functional on membrane compartments in the constitutive secretory pathway. Now, members of the heterotrimeric G protein family have also been localized on intracellular membranes and compartments such as the Golgi complex. We have studied the localization and targeting of G a subunits to distinct membrane domains in polarized epithelial cells. The distribution of different G a subunits on very specific membrane domains in cultured epithelial cells and in epithelial cells of the kidney cortex, is highly suggestive of roles for these G proteins in intracellular trafficking pathways. One of these G protein subunits, Gai-3, was localized on Golgi membranes. Studies on L L C -P K i cells overexpressing Gcti.3 provided evidence for its functional role in regulating the transport o f a constitutively secreted heparan sulfate proteoglycan through the Golgi complex. Inhibition or activation of heterotrimeric G proteins by pertussis toxin or by aluminium fluoride respectively, have provided further evidence for regulation of intracellular transport by pertussis toxin-sensitive G proteins. Although the functions of Golgi-associated G proteins are not yet understood at the molecular level, heterotrimeric G proteins have been implicated in the binding of cytosolic coat proteins and vesicle formation on Golgi membranes. Future studies will elucidate how multiple G proteins, of both the heterotrimeric and monomeric families, are involved in the regulation of Golgi function and protein trafficking in the secretory pathwayArtigo Targeting of chimeric Gαi proteins to specific membrane domains(Journal of Cell Science, 1994) Almeida, Jose Bruno de; Holtzman, Eliezer J.; Peters, Philip; Ercolani, Louis; Ausiello, Dennis A.; Stow, Jennifer L.Heterotrimeric guanine nucleotide-regulatory (G) proteins are associated with a variety of intracellular membranes and specific plasma membrane domains. In polarized epithelial LLC-PK1 cells we have shown previously that endogenous Gαi-2 is localized on the basolateral plasma membrane, whereas Gαi-3 is localized on Golgi membranes. The targeting of these highly homologous Gαi proteins to distinct membrane domains was studied by the transfection and expression of chimeric Gαi proteins in LLC-PK1 cells. Chimeric cDNAs were constructed from the cDNAs for Gαi-3 and Gαi-2 and introduced into a pMXX eukaryotic expression vector containing a mouse metallothioneinI promotor. Stably transfected cell lines were produced that expressed either Gαi-2/3 or Gαi-3/2 chimeric proteins. Chimeric and endogenous Gαi proteins were detected in cells using specific carboxy-terminal peptide antibodies. Immunofluorescence staining was used to localize endogenous and chimeric Gαi proteins in LLC-PK1 cells. The staining of chimeric proteins was detected as an increased intensity of staining on membranes containing endogenous Gαi proteins. Using confocal microscopy and image analysis we localized Gαi-2 to a specific sub-domain of the lateral membrane of polarized cells, the chimeric Gαi-3/2 protein was then shown to colocalize with endognenous Gαi-2 in the same lateral plasma membrane domain. The chimeric Gαi-2/3 protein colocalized with endogenous Gαi-3 on Golgi membranes in LLC-PK1 cells. These results show that chimeric Gαi proteins were targeted to the same membrane domains as endogenous Gαi proteins and the specificity of their membrane targeting was conferred by the carboxy-terminal end of the proteins. These data provide the first evidence for specific targeting information contained in the carboxy termini of Gαi proteins, which appears to be independent of amino-terminal membrane attachment sites in these proteinsArtigo Gravidade da doença renal cística adquirida determina melhora da anemia da insuficiência renal crônica(Revista da Associação Médica Brasileira, 1997) Almeida, José Bruno de; Bezerra, H.M.; Maia, G.P. M.; Bezerra Neto, F. A.; Serra, M.OBJETIVO. Identificar correlação entre duração do tratamento dialítico, gravidade da doença renal cística adquirida (DRCA) medida pelo tamanho dos cistos e seu efeito sobre a correção espontânea da anemia. MATERIAL E MÉTODOS. Foram selecionados dez pacientes, seis do sexo masculino e quatro do feminino, com mais de cinco anos em tratamento dialítico. Nenhum paciente selecionado tinha doença renal policística bilateral como doença primária. A avaliação renal foi feita com ultra-sonografia. DRCA foi caracterizada pela presença de quatro ou mais cistos em cada rim. Os maiores cistos foram medidos para efeito de correlação. O diagnóstico da anemia foi estabelecido pelo valor do hematócrito e da hemoglobina séricos. Foram analisados, também, uréia, creatinina, albumina, ferro, capacidade total de combinação do ferro e o percentual de saturação da transferrina séricos. RESULTADOS. Os pacientes estavam adequadamente dialisados (uréia e creatinina séricas = 98,7 ± 35mg/dL e 9,7 ± 2,7mg/dL, respectivamente), com bom estado nutricional (albumina sérica = 4,5 ± 0,5g/dL), e tinham razoável reserva de ferro (ferro sérico = 80 ± 34mg/dL). A prevalência de DRCA foi de 80%. Não detectamos nenhum sinal ultra-sonográfico de malignidade nesses cistos. Houve correlação significante entre tempo de diálise e hematócrito (R = 0,70; p < 0,05). O tamanho dos cistos teve correlação direta e significante com os valores do hematócrito (R = 0,74; p < 0,05). CONCLUSÕES. Os resultados mostram que a melhora espontânea da anemia observada nos pacientes em diálise crônica se correlaciona de forma significante com a gravidade da doença renal cística adquirida. Isso sugere uma atividade funcional dos cistos renais na produção de eritropoetinaArtigo Myosin II is associated with Golgi membranes: identification of p200 as nonmuscle myosin II on Golgi-derived vesicles(Journal Of Cell Science, 1997) Almeida, José Bruno de; Ikonen, Elina; Fath, Karl R.; Burgess, David R.; Ashman, Keith; Simons, Kai; Stow, Jennifer L.A variety of peripheral membrane proteins associate dynamically with Golgi membranes during the budding and trafficking of transport vesicles in eukaryotic cells. A monoclonal antibody (AD7) raised against Golgi membranes recognizes a peripheral membrane protein, p200, which associates with vesicles budding off the trans-Golgi network (TGN). Based on preliminary findings, a potential association between p200 and myosin on Golgi membranes was investigated. Immunofluorescence staining of cultured cells under a variety of fixation conditions was carried out using an antibody raised against chick brush border nonmuscle myosin II. We show that, in addition to being found in the cytoplasm or associated with stress fibres, nonmuscle myosin II is also specifically localized on Golgi membranes. Myosin II was also detected on Golgi membranes by immunoblotting and by immunogold labeling at the electron microscopy level where it was found to be concentrated on Golgi-derived vesicles. The association of myosin II with Golgi membranes is dynamic and was found to be enhanced following activation of G proteins. Myosin II staining of Golgi membranes was also disrupted by brefeldin A (BFA). Colocalization of the AD7 and myosin II antibodies at the light and electron microscopy levels led us to investigate the nature of the 200 kDa protein recognized by both antibodies. The 200 kDa protein immunoprecipiated by the AD7 antibody was isolated from MDCK cells and used for microsequencing. Amino acid sequence data enabled us to identify p200 as the heavy chain of nonmuscle myosin IIA. In addition, an extra protein (240 kDa) recognized by the AD7 antibody specifically in extracts of HeLa cells, was sequenced and identified as another actin-binding protein, filamin. These results show that nonmuscle myosin II is associated with Golgi membranes and that the vesicle-associated protein p200, is itself a heavy chain of myosin II.Artigo Hipertensão arterial: hipertensão arterial e a progressão da lesão renal. Em que podemos intervir?(Jornal Brasileiro de Nefrologia, 1998) Almeida, José Bruno deEm 1993, cerca de 250.000 pacientes foram tratados para insuficiência renal crônica nos Estados Unidos. Diabetes mellitus (35%), hipertensão arterial (29%) e glomerulonefrite crônica (11%) foram as causas mais comuns da doença renal crônica. Houve um custo anual estimado da ordem de 11 bilhões de dólares. Por esta razão, um dos principais objetivos da nefrologia atual é a redução da incidência de insuficiência renal crônica através da prevenção das nefropatias (prevenção primária) ou retandando a progressão da lesão renal estabelecida (prevenção secundária). 2 A evolução natural da doença renal crônica, independente da etiologia, geralmente é progressivo e culmina com estágio terminal de insuficiência renal crônica e indicação de diálise e/ou transplante renal. Algumas medidas têm sido preconizadas para retardar a progressão da lesão renal, tais como diminuição da ingesta de proteínas, redução da pressão arterial, da excreção urinária de proteínas e dos níveis das lipoproteínas plasmáticas. 3 A proposta desta atualização é revisar os dados da literatura atual sobre a importância do controle da pressão arterial na progressão da lesão renal correlacionando os aspectos fisiopatológicos com as mais recentes evidências clínicasArtigo Effect of end-stage renal disease and diabetes on zinc and copper status(Springer, 2006) Almeida, José Bruno de; Batista, Maria Nazaré; Cuppari, Lílian; Pedrosa, Lucia de Fátima Campos; Almeida, Maria das Graças; Medeiros, Anna Cecília Queiroz de; Canziani, Maria Eugiènia F.The aim of this study was to compare the nutritional status of zinc and copper in patients with and without diabetes submitted to chronic hemodialysis. Thirty-three patients with type 2 diabetes (DM group), 30 nondiabetic patients (NDM group), and 20 healthy individuals (control group) were studied. Plasma, erythrocyte, and urinary zinc and plasma copper were obtained from atomic absorption spectrophotometry and ceruloplasmin by immunonephelometry. The anthropometric parameters were similar among the groups. Plasma zinc was lower and erythrocyte zinc was higher in the DM and NDM groups in relation to the control group. No difference in urinary zinc was observed comparing the groups. Plasma copper was higher in the DM group when compared to the NDM and control groups. Ceruloplasmin was similar in the three groups. Serum urea was a positive independent determinant of plasma zinc concentrations. The determinants of erythrocyte zinc were MAMC midarm nuscle circumference and Kt/V dialysis adequacy. The determinants of plasma copper concentration were serum creatinine and serum glucose. The results of this study demonstrate an alteration in the distribution of zinc of patients with chronic kidney disease (CKD) independently of the presence of DM. Also, the status of copper seems not to be influenced by CKD, but only by the metabolic derangements associated with diabetes.Artigo Oxalose primária diagnosticada após transplante renal - relato de caso(Brazilian Journal of Transplantation, 2007) Almeida, José Bruno de; Costa, Kellen Micheline Alves Henrique; Quinino, Raquel Martins e; Moura, Luiz Antônio Ribeiro de; Félix, Ricardo Humberto de Miranda; Bandeira, Rodrigo de Lima; Silva Júnior, Maurício Ferreira daRelata-se um caso provável de hiperoxalúria primária tipo 1 em uma paciente jovem cujo diagnóstico foi realizado apenas após o transplante renal. O objetivo deste trabalho é chamar atenção para uma enfermidade rara de diagnóstico difícil, principalmente quando se trata de paciente em terapia renal substitutiva. Trata-se de uma adolescente de 16 anos em hemodiálise, que se submeteu a transplante renal com doador vivo relacionado (mãe), inicialmente com boa resposta do enxerto, chegando a apresentar creatinina de 1,2mg/dl no 2º dia pós- operatório (DPO), porém com piora progressiva e rápida da função renal no 4º DPO. Realizou-se biópsia renal e a oxalose primária foi diagnosticada. Apesar do tratamento instituído com Piridoxina e diálise diária, houve perda do enxerto. O tratamento definitivo da doença consiste no transplante hepatorrenal, devendo este ser realizado o mais precocemente possível. O conhecimento dessa enfermidade por parte dos nefrologistas é relevante, uma vez que a paciente poderia ter sido mais bem preparada para o transplante renal, se tivesse chegado ao ambulatório com esse diagnóstico.Artigo Infecção por polioma vírus após transplante renal: relato de caso(Brazilian Journal Of Transplantation, 2010) Almeida, Jose Bruno de; Costa, Kellen Micheline Alves Henrique; Félix, Ricardo Humberto de Miranda; Silva Júnior, Maurício Ferreira da; Franco, Marcelo Fabiano deA nefropatia por poliomavírus ocorre em 1,1% a 9,3% dos pacientes transplantados renais, causando perda do enxerto em mais de 40% dos casos. A infecção primária ocorre durante a infância e o vírus se mantém latente no organismo, sendo reativado em estados de imunossupressão. Relatamos o caso de uma mulher de 45 anos transplantada renal há três anos, em uso crônico de tacrolimo e micofenolato mofetil, que apresentou perda progressiva da função renal durante episódio diarréico, mesmo com tratamento adequado. Biópsia do enxerto renal revelou presença de células com inclusões por poliomavírus, sendo iniciado sirolimo e imunoglobulina humana. Com a terapêutica instituída, houve melhora da função renal e nova biópsia realizada após a internação não demonstrou alterações histológicas de poliomavírusArtigo Pseudotumor cerebral associado ao uso de ciclosporina após transplante renal(Jornal Brasileiro de Nefrologia, 2010) Almeida, Jose Bruno de; Costa, Kellen Micheline Alves Henrique; Félix, Ricardo Humberto de Miranda; Silva Júnior, Maurício Ferreira daPseudotumor cerebral (PC) é uma síndrome, caracterizada pela presença de hipertensão intracraniana (HIC) e sistema ventricular normal. Pacientes submetidos a transplante renal parecem ser mais suscetíveis a desenvolvê-la, devido à terapia com imunossupressores. Ciclosporina (CsA) é uma causa rara de PC, pouco descrita na literatura e que deve ser lembrada no diagnóstico diferencial de HIC e papiledema nesses pacientes. Relatamos um caso de um menino de 10 anos, há três anos com enxerto renal, em uso crônico de micofenolato mofetil (MMF), CsA e baixas doses de prednisona que apresentou quadro de cefaleia, vômitos, diplopia e fotofobia. Fundoscopia revelou edema de papila bilateral. Exame do líquor (LCR) e de imagem foram normais. Após exclusão de causas secundárias, foi feito diagnóstico de PC devido ao uso crônico de CsA, que, portanto, foi substituída por Sirolimus. O paciente apresentou melhora clínica progressiva, com resolução do papiledema após três meses