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D Blanc

Publications and source records attributed to D Blanc.

At least 37 records · Page 2Linked to original sources

Human immunodeficiency virus type 1 Vif protein binds to the Pr55Gag precursor.

The Vif protein of human immunodeficiency virus type 1 is required for productive replication in peripheral blood lymphocytes. Previous reports suggest that vif-deleted viruses are limited in replication because of a defect in the late steps of the virus life cycle. One of the remaining questions is to determine whether the functional role of Vif involves a specific interaction with virus core proteins. In this study, we demonstrate a direct interaction between Vif and the Pr55Gag precursor in vitro as well as in infected cells. No interaction is observed between Vif and the mature capsid protein. The Pr55Gag-Vif interaction is detected (i) in the glutathione S-transferase system, with in vitro-translated proteins demonstrating a critical role of the NC p7 domain of the Gag precursor; (ii) with proteins expressed in infected cells; and (iii) by coimmunoprecipitation experiments. Deletion of the C-terminal 22 amino acids of Vif abolishes its interaction with the Pr55Gag precursor. Furthermore, point mutations in the C-terminal domain of Vif which have been previously shown to abolish virus infectivity and binding to cell membranes dramatically decrease the Gag-Vif interaction. These results suggest that the interaction between Vif and the pr55Gag precursor is a critical determinant of Vif function.

Amino Acid Sequence↗

Phenotypically Vif- human immunodeficiency virus type 1 is produced by chronically infected restrictive cells.

The permissivity of CD4+ transformed T cells for the replication of human immunodeficiency virus type 1 (HIV-1) vif mutants varies widely between different cell lines. Mutant vif-negative viruses propagate normally in permissive CD4+ cell lines but are unable to establish a productive infection in restrictive cell lines such as H9. As a consequence, elucidation of the function of Vif has been considerably hampered by the inherent difficulty in obtaining a stable source of authentically replication-defective vif-negative viral particles produced by restrictive cells. vif-negative, vpr-negative HIV-1 strain NDK stock, produced by the permissive SupT1 cell line, was used to infect restrictive H9 cells. By using a high multiplicity, infection of H9 cells was achieved, leading to persistent production of viral particles displaying a dramatically reduced infectious virus titer when measured in a single-cycle infectivity assay. Although these viral particles were unable to further propagate in H9 cells, they could replicate normally in CEM and SupT1 cells. Comparison of unprocessed and processed Gag proteins in the persistently produced vif-negative viral particles revealed no defect in the processing of polypeptide precursors, with no inversion of the Pr55gag/p24 ratio. In addition, there was no defect in Env incorporation for the vif-negative viral particles. Despite their apparently normal protein content, these particles were morphologically abnormal when examined by transmission electron microscopy, displaying a previously described abnormally condensed nucleoid. Chronically infected restrictive cell lines producing stable levels of phenotypically vif-negative HIV-1 particles could prove particularly useful in further studies on the function of Vif in the virus life cycle.

Cell Line↗

Peripheral blood mononuclear cells produce normal amounts of defective Vif- human immunodeficiency virus type 1 particles which are restricted for the preretrotranscription steps.

Previous studies have demonstrated the absence of viral replication of Vif- mutants in stimulated primary blood mononuclear cells (PBMC). Human immunodeficiency virus type 1 strain NDK Vif- mutants were propagated on the semipermissive CEM cell line, and the viral stock obtained was compared with the wild-type virus during a single cycle in PBMC. The Vif- virus was able to enter PBMC with the same efficiency as the wild type, as demonstrated by quantification of the strong-stop cDNA, and retrotranscription was observed for both viruses within 4 h postinfection. Using a PCR assay with an Alu-long terminal repeat pair of primers, we detected integration for both the wild-type and Vif- viruses. We then used qualitative and quantitative reverse transcription-mediated PCR techniques to study the steady-state level of intracellular and extracellular viral RNAs. All mRNA species were detected in PBMC infected with the wild-type virus or with the Vif- virus 36 h postinfection. Furthermore, quantification of viral RNA released from infected cells demonstrated similar levels of virus produced after a unique cycle of replication. However, the Vif- virus obtained after one replication cycle in PBMC was unable to initiate retrotranscription in permissive target cells. These data strongly suggest that the failure to infect target cells is due to a defect in the formation of the viral particle in PBMC.

Base Sequence↗

Nosocomial pneumonia in mechanically ventilated patients receiving antacid, ranitidine, or sucralfate as prophylaxis for stress ulcer. A randomized controlled trial.

OBJECTIVE: To assess three anti-stress ulcer prophylaxis regimens in mechanically ventilated patients for bacterial colonization, early- and late-onset nosocomial pneumonia, and gastrointestinal bleeding. DESIGN: Randomized controlled trial. PATIENTS: Consecutive eligible patients with mechanical ventilation and a nasogastric tube. Of 258 eligible patients, 244 were assessable. SETTING: Medical and surgical intensive care units. INTERVENTION: At intubation, patients were randomly assigned to receive one of the following: antacid (a suspension of aluminum hydroxide and magnesium hydroxide), 20 mL every 2 hours; ranitidine, 150 mg as a continuous intravenous infusion; or sucralfate, 1 g every 4 hours. MEASUREMENTS: Using predetermined criteria, the incidence of gastric bleeding, gastric colonization, early-onset pneumonia, and late-onset pneumonia was assessed in patients intubated for more than 24 hours. RESULTS: Of 244 assessable patients, macroscopic gastric bleeding was observed in 10%, 4%, and 6% of patients assigned to receive sucralfate, antacid, and ranitidine, respectively (P > 0.2). The incidence of early-onset pneumonia was not statistically different among the three treatment groups (P > 0.2). Among the 213 patients observed for more than 4 days, late-onset pneumonia was observed in 5% of the patients who received sucralfate compared with 16% and 21% of the patients who received antacid or ranitidine, respectively (P = 0.022). Mortality was not statistically different among the three treatment groups. Patients who received sucralfate had a lower median gastric pH (P < 0.001) and less frequent gastric colonization compared with the other groups (P = 0.015). Using molecular typing, 84% of the patients with late-onset gram-negative bacillary pneumonia were found to have gastric colonization with the same bacteria before pneumonia developed. CONCLUSION: Stress ulcer prophylaxis with sucralfate reduces the risk for late-onset pneumonia in ventilated patients compared with antacid or ranitidine.

Adult↗

[What is the dose?].

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History, 19th Century↗

Transcomplementation of VIF- HIV-1 mutants in CEM cells suggests that VIF affects late steps of the viral life cycle.

The vif gene of HIV-1 has previously been claimed to be essential for the ability of cell-free virus preparations to infect cells. Here we report that the CEM T-cell-line, stably transfected with and expressing vif, supports the replication of vif- HIV-1 viruses to the same extent as wild-type HIV1. Cell entry and early replication stages are the same for vif- and vif+ HIV-1 passaged in CEM, as measured both by a PCR-based cell entry assay and by fusogenic potential. These findings indicate that vif does not affect viral infectivity on CEM cells, but seems to act at a later stage of virus replication/maturation. We also show that the VIF proteins of two different HIV-1 strains can transcomplement different vif- HIV-1 mutants.

Cell Line↗

Human and simian immunodeficiency viruses: virus-receptor interactions.

The major cellular receptor for the primate immunodeficiency viruses is the CD4 molecule. As well as mediating virion attachment to the cell surface, CD4 is thought to activate the viral fusion pathway. CD4 is not, however, sufficient for viral entry; other molecules are probably involved, and in certain circumstances these may substitute for CD4. Viral tropism and cytopathogenicity are also influenced by receptor interactions.

Amino Acid Sequence↗

[Calcifying panniculitis].

We report a case of calcifying panniculitis which occurred in a 67-year old woman suffering from chronic renal failure and primarily involved anticoagulant injection sites. Calcifying panniculitis is a rare condition belonging to the spectrum of calciphylaxis first described by Selye in 1962. This disease seems to imply necessarily a specific morbid background including renal failure together with calcium-phosphate metabolism impairment. Following the action of a so-called "challenger" (in the present case calcium heparinate [Calciparine], a subcutaneously administered anticoagulant drug), the lesions start as painful subcutaneous nodules soon turning into plaques of necrosis. The area involved mainly includes the abdomen and thighs, due to the preferential distribution of fat and the classical use of such regions for subcutaneous injections. In our patients, histological examination disclosed an initial involvement of subcutaneous fat vessels mimicking mediacalcosis, soon followed by calcium-phosphate deposits within interadipocyte spaces. An electron microscopy study confirmed the presence of calcium crystals within the cytoplasms of some connective tissue cells and the extracellular matrix. From a review of the literature, it seems reasonable to normalize the calcium-phosphate product by reducing hyperparathyroidism as a first therapeutic step. Then, wide surgical excision of the necrotic areas should be performed to rule out any possibility of self worsening of the lesions and provide the best chance of healing without superinfection. The use of diphosphonates on pathophysiological grounds is discussed.

Aged↗

[Abrikossof's tumor of the esophagus. An original case. Review of the literature].

Granular-cell tumors (GCT), also called Abrikossof's tumors, are generally benign, ubiquitous tumors. An original case of granular-cell tumor of the esophagus is reported. The symptoms included dysphagia and pyrosis. Fiberendoscopy showed a peptic esophageal stenosis with ulcerations confirmed by biopsy. CT showed a round thickening of the esophageal wall, localized in height. This lesion had previously been the object of several dilatation attempts. Esophagectomy with esogastric anastomosis in the thorax was performed. The histological study allowed diagnosing a granular-cell tumor, though one of a very peculiar type: misleading symptoms, tumor infiltrating the whole height of the esophagus and a circular area. Ninety cases of granular-cell tumors are reported in the literature; they are associated with a cancer of the air passages or of the digestive tract in 11% of cases. The cancer often appears secondarily, which requires lengthy surveillance. We do not know if this association is directly related or accidental. We have found no identical case of such a lesion, involving both the whole circumference and the whole height of the esophagus, in the literature.

Esophageal Neoplasms↗

[Colonic perforation during colonoscopy. 100 cases].

The analysis of 100 cases of colon perforation during colposcopic examinations highly demonstrates such a statement. The perforation risk during colposcopies is generally of the order of 0.2% for a diagnosis coloscopy. According to the statistic data used, it can reach 0.5 to 3% in therapy coloscopy. This is a risk inherent to the technique used. It is thus required to analyse the causes and take the appropriate measures to reduce it to a minimum. Mortality due to such a complication remains high (14%), i.e about 0.015 to 0.1% (#2/10000) of all colposcopies. In 11% of the patients, serious sequelae are to be observed. This demonstrates the significance of the medico-legal problem set by these perforations during colposcopies. The whole personnel responsibility can be involved: colposcopist, surgeon, anesthetist and hospital unit.

Adult↗

Entamoeba histolytica zymodemes: exhibition of gamma and delta bands only of glucose phosphate isomerase and phosphoglucomutase may be influenced by starch content in the medium.

Entamoeba histolytica isolated from human can be associated with either symptomatic disease or with asymptomatic carriers. Pathogenic and nonpathogenic strains can be distinguished on the basis of differences in the electrophoretic patterns of four isoenzymes (zymodeme). With glucose phosphate isomerase and phosphoglucomutase, we observed variation of the expression of their gamma bands as a function of starch volume in culture. We cultured E. histolytica strains from different zymodemes in Robinson medium using both a low (2-4 mg/bottle) and a high (12-15 mg/bottle) content of rice starch as supplement. These cultures were monitored by electrophoresis of glucose phosphate isomerase and phosphoglucomutase. Strains having gamma or delta bands exhibited those bands when a high content of starch was used in culture, but did not do so with a low content. Contrarily, strains that never exhibited those bands did not express them when the amount of starch in the culture was increased. However, alpha and beta bands of the same isoenzymes were always present and never showed any variation. The results suggest that expression of gamma and delta bands of glucose phosphate isomerase and phosphoglucomutase are subject to culture conditions and that genes coding for those isoenzymes may be different from those coding for the alpha and beta bands.

Animals↗

Specific cell surface requirements for the infection of CD4-positive cells by human immunodeficiency virus types 1 and 2 and by Simian immunodeficiency virus.

Human CD4 was expressed on a range of mammalian cell lines. CD4+ non-primate cells, derived from rat, hamster, mink, cat, and rabbit, bind recombinant gp120 of human immunodeficiency virus type 1 (HIV-1) but are resistant to HIV-1 infection. CD4 expression on various human, rhesus, and African green monkey cell lines confers differential susceptibilities for HIV-1, HIV-2, and simian immunodeficiency (SIV) strains. For example, CD4+ TE671 rhabdomyosarcoma cells are sensitive to HIV-1 and HIV-2 but resistant to SIV, whereas CD4+ U87 glioma cells are resistant to HIV-1 infection but sensitive to HIV-2 and SIV. HIV-1 infection was not dependent on human major histocompatibility class I expression. Studies of cell fusion and of infection by vesicular stomatitis virus pseudotypes bearing HIV-1 and HIV-2 envelopes showed that the differential cell tropisms of HIV-1, HIV-2, and SIV are determined at the cell surface.

Acquired Immunodeficiency Syndrome↗