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M A Costa de Beauregard

Publications and source records attributed to M A Costa de Beauregard.

8 recordsLinked to original sources
PubMed2000

Functional cystic fibrosis transmembrane conductance regulator tagged with an epitope of the vesicular stomatis virus glycoprotein can be addressed to the apical domain of polarized cells.

The cystic fibrosis transmembrane conductance regulator (CFTR) is a phosphorylation-activated chloride channel apically localized in epithelial cells. In cystic fibrosis patients, the gene encoding this N-linked glycoprotein is mutated. About 70% of CF patients express a mutated form of CFTR, deleted at the phenylalanine residue at position 508 (deltaF508). CFTR-deltaF508 fails to exit the endoplasmic reticulum; it remains incompletely glycosylated and is rapidly degraded. To optimize CFTR detection for membrane localization studies and biochemical studies, we tagged wild-type and deltaF508 CFTR with the VSV-G epitope at their carboxy-terminal ends. We have generated pig kidney epithelial cell clones (LLCPK1) expressing VSV-G-tagged human wild-type and deltaF508-CFTR. In CFTR-expressing cells, the transfected protein is maturated and transported to the apical membrane where it is concentrated. The cells exhibit a strong anion channel activity after stimulation by cAMP, as demonstrated by a halide sensitive fluorescent dye assay (6-methoxy-N-ethylquinominium, SPQ), and whole-cell patch-clamp approach. This activity of CFTR-VSV-G is indistinguishable from the wild-type CFTR. In contrast, in cells expressing tagged deltaF508-CFTR or in non-transfected cells, no anion channel activity could be detected after stimulation by cAMP. In deltaF508-CFTR-VSV-G-expressing cells, the mutated CFTR remained in the incompletely glycosylated form and was localized in the endoplasmic reticulum. These cell lines reproduce the cellular fate of wild-type and mutated CFTR-deltaF508. To our knowledge, they are the first differentiated epithelial cell lines stably expressing tagged CFTR and CFTR-deltaF508 in which cellular processing and functional activity of these two proteins are reproduced. Thus the addition of the VSV-G epitope does not impair the localization and function of CFTR, and these cell lines can be used to examine CFTR function in vitro.

Cell Line↗
PubMed2000

Evaluation of FDG uptake by renal malignancies (primary tumor or metastases) using a coincidence detection gamma camera.

UNLABELLED: The aim of this study was to evaluate the usefulness of FDG scanning using an ordinary gamma camera equipped with coincidence detection (CDET) for 2 renal cancer indications: characterization and staging of renal masses before nephrectomy and search for recurrence after nephrectomy. METHODS: Between September 1997 and June 1998, a whole-body scan and at least 1 tomoscintigram were obtained on 23 occasions in 22 patients (fasting for at least 6 h) using a Prism XP 2000 CDET gamma camera; scanning was begun 45 min after intravenous injection of 150-250 MBq FDG. RESULTS: Postoperative histologic evidence was obtained from 13 of 16 patients who underwent FDG using a CDET gamma camera before renal surgery; 4 renal masses did not accumulate FDG (3 true-negatives, 1 false-negative), whereas 9 renal tumors accumulated FDG (8 true-positives, 1 false-positive). In the other 3 patients, only 1 extrarenal site of FDG uptake was checked and confirmed on histologic examination: a bone metastasis from renal cell carcinoma in 2 cases and lymph node metastasis from a squamous cell carcinoma (3 true-positives). The primary local and regional staging of the malignant renal tumors was accurate in the 9 patients who underwent nephrectomy (8 true-negatives, 1 true-positive). The primary distant staging was positive in 1 case (focus in the chest corresponding to a probable true-positive on follow-up). In the 7 examinations performed because of suspected recurrence of renal cell carcinoma several months after nephrectomy, metastases were visualized by FDG in 4 patients, confirmed by biopsy in 2 patients, and confirmed by conventional imaging or follow-up (or both) in 2 patients. The other 3 patients had negative FDG scans, corresponding to probable true-negative results on follow-up. CONCLUSION: FDG using a CDET gamma camera can be used effectively for the staging and restaging of renal tumors and might be useful for characterization of the primary renal tumor in doubtful cases.

Carcinoma, Renal Cell↗
PubMed1999

Aspirin and some other nonsteroidal anti-inflammatory drugs inhibit cystic fibrosis transmembrane conductance regulator protein gene expression in T-84 cells.

Cystic fibrosis (CF) is caused by mutations in the CF gene, which encodes CF transmembrane conductance regulator protein (CFTR), a transmembrane protein that acts as a cAMP-regulated chloride channel The disease is characterized by inflammation but the relationship between inflammation, abnormal transepithelial ion transport, and the clinical manifestations of CF are uncertain. The present study was undertaken to determine whether three nonsteroidal anti-inflammatory drugs (NSAIDs) (aspirin, ibuprofen, and indomethacin) modulate CFTR gene expression in T-84 cells. Treatment with NSAIDs reduced CFTR transcripts, and decreased cAMP-stimulated anion fluxes, an index of CFTR function. However, the two phenomena occurred at different concentrations of both drugs. The results indicate that NSAIDs can regulate both CFTR gene expression and the function of CFTR-related chloride transport, and suggest that NSAIDs act via multiple transduction pathways.

1-Methyl-3-isobutylxanthine↗
PubMed1998

Bacterial endocarditis associated with crescentic glomerulonephritis in a kidney transplant patient: first case report.

BACKGROUND: Endocarditis-induced crescentic glomerulonephritis is a well-described complication in nontransplant patients. Its occurrence in transplant patients has not been reported to date. METHODS: A 50-year-old man who had received a renal allograft 13 years before and been treated with prednisone, 10 mg/day, was admitted for progressive renal failure, purpura, edema of the lower limbs, and fever. RESULTS: Blood cultures isolated Streptococcus bovis and cardiac ultrasound examination revealed a 23-mm-large vegetation on the mitral valve. His plasma creatinine level was 478 micromol/L and his proteinuria was 5.5 g/day. A renal biopsy showed diffuse crescentic glomerulonephritis. Long-term antibiotic treatment and three methylprednisolone pulses were effective in treating the endocarditis and glomerulonephritis. CONCLUSION: Endocarditis-induced glomerulonephritis is an immune-mediated disease that can also occur on a renal allograft. It is likely that a low daily dose of immunosuppressive treatment may have been a facilitating factor.

Creatinine↗
PubMed1995

Suppression of villin expression by antisense RNA impairs brush border assembly in polarized epithelial intestinal cells.

We have used an antisense RNA strategy to investigate the role of the actin-associated protein, villin, in the brush-border morphogenesis of human intestinal CaCO2 cells. Stable expression of a cDNA encoding antisense villin RNA resulted in the permanent down-regulation of the endogenous villin message and dramatically affected brush-border assembly. Ultrastructural and immunolocalization studies revealed that epithelial cell polarity was largely maintained. However, in contrast to brush-border markers such as dipeptidyl-peptidase IV, the apical localization of sucrase-isomaltase was specifically impaired. Retransfection of the villin antisense-expressing cell line with a cDNA encoding a partial sense villin RNA restored both brush-border assembly and sucrase-isomaltase apical expression. The suggestion that brush-border morphogenesis may be important for the trafficking of certain proteins is discussed.

Actins↗