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Biomedical subjects

N Robert

Publications and source records attributed to N Robert.

49 records · Page 3Linked to original sources

Rapid decrease in the levels of the double-stranded RNA-dependent protein kinase during virus infections.

The double-stranded RNA-dependent protein kinase from human cells is a 68,000 molecular weight protein (p68 kinase), the level of which is enhanced significantly in cells treated with interferon. With a monoclonal antibody specific for p68 kinase, here we show the phosphorylation and steady-state levels of p68 kinase during virus infection. The p68 kinase is phosphorylated in interferon-treated cells during infection with encephalomyocarditis virus (EMCV), vesicular stomatitis virus (VSV), and vaccinia virus, thus indicating activation of p68 kinase during these virus infections, an essential step required for autophosphorylation of p68 kinase. However, in spite of this activation, the level of p68 kinase is rapidly decreased in virus-infected cells. The half-life of p68 kinase in uninfected cells is 6 to 7 hr, whereas in EMCV-infected cells it is 2 to 3 hr. This decrease in the level of p68 kinase is dependent on the multiplicity of virus infection and it seems to be specific since other cellular proteins as well as the activity of 2'-5'-oligoadenylate synthetase are not modified. Decreased levels of p68 kinase are also observed in cells infected with VSV and vaccinia virus. In the absence of virus infection, decreased levels of p68 kinase occur in cells following incubation with poly(I).poly(C).

2',5'-Oligoadenylate Synthetase↗

Study of anti-Fya in five black Fy(a-b-) patients.

Fya antibodies were detected in 5 Fy(a-b-) black patients during a 5-year period. The Duffy phenotype was confirmed using anti-Fya, anti-Fyb, and anti-Fy3 by an adsorption-elution technique. These 5 patients had sickle cell disease and had received multiple blood transfusions. The antibodies were all found during the investigation of a hemolytic transfusion reaction or failure to achieve the desired red cell increments after transfusion.

Adult↗

[Hypertensive retinopathy and papilledema. Apropos of a case of glycyrrhizin poisoning].

We present a case report of a patient suffering of severe hypertension and grade IV hypertensive retinopathy according to the Keith-Wagener classification. Simultaneously hypertensive encephalopathy was present confirming the poor prognosis of discoedema occurring in the course of hypertension when left untreated. Hypertension was related to chronic poisoning with glycyrrhiza, and its hyperaldosteronism like effect. Pathogenesis of discoedema in malignant hypertension is discussed.

Adult↗

Enhanced level of double-stranded RNA-dependent protein kinase in peripheral blood mononuclear cells of patients with viral infections.

The protein kinase activity dependent on double-stranded RNA was assayed in extracts of peripheral blood mononuclear (PBM) cells from healthy volunteers and in patients with different types of virus infections. The protein kinase was assayed after one-step purification on an immunoaffinity column containing monoclonal antibody against the 68,000 Mr protein, a subunit of the protein kinase. In healthy individuals, the activity of the protein kinase remains constant. In contrast, the activity of the protein kinase is enhanced significantly in patients with viral infections and is decreased during the course of the disease in parallel with clinical ameliorations and reversal of clinical symptoms. There is a strong correlation between the enhanced levels of the protein kinase activity and another interferon-mediated enzyme, 2-5A synthetase. Both of these enzymes, therefore, could be used as markers to evaluate the state of the disease and recovery. In the different populations of lymphocytes, most of the protein kinase activity was found to be present in T and B lymphocytes, T lymphocytes showing a higher activity than B lymphocytes.

2',5'-Oligoadenylate Synthetase↗

Continuous production of interferon in normal mice: effect of anti-interferon globulin, sex, age, strain and environment on the levels of 2-5A synthetase and p67K kinase.

Two interferon-mediated enzymes, a 2-5A synthetase and a kinase that phosphorylates a 67 000 mol. wt. (p67K) protein were found at variable levels in different organs of mice. Among the different strains of mice included in this study, germ-free mice had the lowest levels of these enzymes. The levels of 2-5A synthetase and p67K kinase were enhanced significantly in all mice following treatment with mouse (alpha + beta) interferon. Here, we show that the presence of 2-5A synthetase and p67K kinase in different organs of normal mice (untreated) was due, at least in part, to a constant production of interferon under different physiological conditions. Accordingly, injection of normal mice with anti-mouse interferon (alpha + beta) globulin led to a significant decrease in the level of 2-5A synthetase and p67K kinase. In conventional mice (C3H/He), the level of both of these enzymes was higher in female than in male animals and was decreased with age or when such animals were reared isolated in a pathogen-free protected unit. The levels of 2-5A synthetase and p67K kinase were also decreased in normal mice following injection with a powerful antibiotic against a very wide spectrum of Gram-positive and Gram-negative bacteria. These results suggest that the production of interferon was induced continuously in normal mice. Such induction was mediated by both internal and external agents.

2',5'-Oligoadenylate Synthetase↗

Use of an anti-human leukocyte interferon monoclonal antibody for the purification and radioimmunoassay of human alpha interferon.

Mouse monoclonal antibody directed against human leukocyte alpha interferon (IFN-alpha) was coupled to Sepharose and used as an immunoadsorbent to purify human IFN-alpha. Leukocyte and lymphoblastoid (Namalva) IFNs were retained by the immunoadsorbent with a specificity of 80 to 100% and 40 to 60%, respectively. Human IFN-beta or -gamma and mouse IFN were not retained. The purified IFN-alpha retained its antiviral and anticellular properties as well as its ability to induce the 2-5A synthetase in human cells with a specific activity similar to that of the crude IFN. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radioactively labeled IFN showed that it consisted of several proteins in the molecular weight range of 17,000 to 27,000. 125I-labeled IFN-alpha with a high specific activity (2,000 Ci/mmol) was used in a radioimmunoassay for the titration of IFN-alpha.

Antibodies, Monoclonal↗

Assay and the levels of 2-5A-synthetase in lymphocytes of patients with viral, bacterial and autoimmune diseases.

The level of 2-5A-synthetase in extracts of peripheral blood lymphocytes was estimated by the capacity of the enzyme to synthesize 2-5A in the presence of ATP. The 2-5A was then purified on a small column of DEAE-cellulose. Here we show that under or experimental conditions, the amount of 2-5A formed is proportional to the level of the enzyme. The concentration of 2-5A (nmol of AMP/10(6) lymphocytes) in an assay therefore reflects the level of the enzyme. Enhanced levels of 2-5A synthetase were observed in the lymphocytes of patients with viral and bacterial infections. In most of the cases studied, these enhanced levels of the enzyme decreased during the course of infection parallel to recovery. Thus, the level of 2-5A synthetase may indicate the state of the disease and its evolution during the period of treatment; furthermore, it may be a useful marker in monitoring the return to a normal physiological condition. In addition to patients with viral and bacterial infections, enhanced levels of 2-5A synthetase were observed in patients suffering from autoimmune diseases. In five patients studied here, the enhanced levels of 2-5A synthetase remained high at different periods during the course of the disease. These results suggest the presence of circulating interferon throughout the disease.

2',5'-Oligoadenylate Synthetase↗