Search PubMed⌕ Search

Biomedical subjects

H Jacobsen

Publications and source records attributed to H Jacobsen.

At least 91 records · Page 5Linked to original sources
PubMed1985

Studies on interferon-sensitive cells derived from the interferon-resistant NIH 3T3 clone 1 line.

From the NIH 3T3 clone 1 line which is normally unprotected by interferon (IFN) against lytic virus infection we have selected subclones which show high sensitivity to IFN. The selection procedure was based on encephalomyocarditis virus (EMCV) as selection agent. In the IFN-sensitive subclones thus obtained EMCV replication was inhibited by IFN to a similar degree as observed in L929 cells. Like in the original NIH 3T3 clone 1 line, however, replication of vesicular stomatitis virus (VSV) and cell multiplication were only marginally affected by IFN. We measured the levels of known IFN-induced enzymes (2-5A-synthetase, dsRNA protein kinase and 2-5A-dependent RNase) in a number of subclones and found no consistent differences to the original population. Thus, the newly acquired IFN-dependent protection against EMCV may be mediated by a different antiviral mechanism.

2',5'-Oligoadenylate Synthetase↗
PubMed1983

A sensitive immunoenzymometric assay for 2',5'-oligoadenylate. Detection of elevated 2',5'-oligoadenylate synthetase in human peripheral mononuclear cells.

A competition immunoenzymometric assay for 2',5'-oligoadenylate was developed and employed to measure the interferon-inducible enzyme 2',5'-oligoadenylate synthetase in cell extracts. Microtiter plates coated with a novel conjugate of p5'A2'p5'A2'p5'A and N-(2-aminoethyl)-carbamylmethylated-Ficoll (AECM-Ficoll) bound rabbit polyclonal or mouse monoclonal antibody directed against 2',5'-oligoadenylate. Binding was inhibited by soluble 2',5'-oligoadenylate. Estimates of 2',5'-oligoadenylate concentrations based on inhibition of antibody binding compared favorably with those obtained using a protein synthesis inhibition assay. Low concentrations of 2',5'-oligoadenylate synthesized in vitro by extracts of human peripheral mononuclear cells were conveniently estimated using less than or equal to 10(6) cells. Virtually identical results were obtained when either total extract or synthetase bound to poly(I) . poly(C)-agarose was used for the in vitro incubation. When peripheral mononuclear cells were incubated in vitro with interferon, the normally low levels of 2',5'-oligo(A) synthetase rose dramatically. The assay was employed to measure synthetase levels in peripheral mononuclear cells isolated from patients with systemic lupus erythematosus. Some of these patients were found to have elevated levels of 2',5'-oligoadenylate synthetase.

2',5'-Oligoadenylate Synthetase↗
PubMed1983

Interferon-induced synthesis of 2-5A-dependent RNase in mouse JLS-V9R cells.

Interferon treatment of mouse JLS-V9R cells resulted in a 10- to 20-fold increase in the levels of the 2-5A (ppp(A2'p)nA)-dependent RNase. The nuclease was monitored in cell extracts by covalent and noncovalent binding of 32P-labeled 2-5A derivatives to the nuclease and by the appearance of 2-5A-mediated ribosomal RNA cleavage products. Untreated JLS-V9R cells contained very low levels of the nuclease (5-10% of that found in control Ehrlich ascites tumor cells). An increase in the nuclease was first detected between 3 and 6 hr after interferon treatment and was completely inhibited by actinomycin D. Control experiments indicated that the increase in the nuclease was not due to an intracellular redistribution of the enzyme. The results indicate that in JLS-V9R cells, interferon controls the activity of the 2-5A system at two levels: by inducing the synthesis of both the 2-5A-dependent RNase and the 2-5A-synthetase.

2',5'-Oligoadenylate Synthetase↗
PubMed1983

Double-stranded RNA-dependent phosphorylation of protein P1 and eukaryotic initiation factor 2 alpha does not correlate with protein synthesis inhibition in a cell-free system from interferon-treated mouse L cells.

The double-stranded RNAs (I)n X (C)n and (A)n X (dUfl)n (dUfl is 2'-fluoro-2'-deoxyuridylic acid) have been compared as inhibitors of translation in cell-free systems from interferon-treated mouse L cells and from rabbit reticulocytes. In the interferon-treated mouse L-cell system, both double-stranded RNAs stimulated kinase activity, leading to phosphorylation of protein P1 and eukaryotic initiation factor 2 alpha (eIF-2 alpha), but only (1)n X (C)n activated the (2'-5')-oligoadenylate synthetase. Moreover, in this system, (I)n X (C)n, but not (A)n X (dUfl)n, inhibited translation. Both (A)n X (dUfl)n and (I)n X (C)n also activated the rabbit reticulocyte kinase to phosphorylate protein P1 and eIF-2 alpha, but, in contrast to mouse L-cell systems, both (A)n X (dUfl)n and (I)n X (C)n were potent inhibitors of translation in reticulocyte lysates. These results indicate that protein P1 and eIF-2 alpha phosphorylation are not sufficient to cause inhibition of protein synthesis in interferon-treated mouse L-cell extracts. They further suggest that protein synthesis inhibition by (I)n X (C)n in extracts of interferon-treated L cells correlates better with activation of (2'-5')-oligoadenylate synthetase than with activation of the protein P1 and eIF-2 alpha kinase.

Animals↗
PubMed1983

Induction of ppp(A2'p)nA-dependent RNase in murine JLS-V9R cells during growth inhibition.

We recently reported that interferon induces the synthesis of ppp(A2'p)nA(n = 2 to greater than or equal to 4) (2-5A)-dependent RNase in the murine cell line JLS-V9R. These cells normally contain very low levels of the nuclease; after interferon treatment, however, they develop levels approaching those found in murine L or Ehrlich ascites tumor cells. Here, we report a similar increase in the nuclease levels in JLS-V9R cells during the transition from the subconfluent actively growing state to the confluent stationary phase. Levels of 2-5A synthetase increased in parallel with the nuclease. The induced levels of both the nuclease and synthetase returned to low basal amounts after trypsinization, dilution, and culturing of the cells at subconfluent densities. The addition of anti-murine interferon (alpha + beta) antibodies to the medium did not affect the induction of the nuclease nor could any interferon be detected in the culture supernatants as determined by the lack of antiviral activity. The increase in the enzymes was not, therefore, due to the spontaneous production of interferon. The induction of the nuclease during confluency preceded an inhibition of [3H]-thymidine incorporation by the cells into DNA. The regulation of the 2-5A-dependent RNase in JLS-V9R cells may, therefore, be related to the control of cell growth.

2',5'-Oligoadenylate Synthetase↗
PubMed1982

Spontaneous spatiotemporal organization in yeast extracts.

Stirred cytoplasmic extracts of the yeast Saccharomyces uvarum can, on addition of trehalose, display oscillations in the metabolite concentrations. Unstirred extracts may also develop spatiotemporal structures expressed in the metabolite concentrations. In this paper we present evidence of the existence of such patterns. The latter were recorded using the optical absorption of NADH in an annular cuvette, which was rotated in a photometer. Regularly spaced wavelike spatiotemporal standing waves were observed. At certain fixed points the NADH concentration appeared to remain constant. Such phenomena may improve our understanding of biochemical morphogenesis.

Glycolysis↗
PubMed1982

The clinical effect and the effect on serum IgG antibodies to peripheral nerve tissue of plasma exchange in patients wit Guillain-Barré syndrome.

The mixed haemagglutination technique was used to demonstrate IgG antibodies to peripheral nerve tissue in sera from patients with the Guillain-Barré syndrome. The clinical effect and the effect on the antibodies of plasma exchange were examined in 18 patients. Neurological examination with muscle testing and neurophysiological examination of the patients were performed before and immediately after plasma exchange. Before the exchange antibodies were detected in sera from 11 of the patients. These patients showed clinical improvement during the treatment. After plasma exchange, antibodies were detected in sera from only two of the patients. The seven patients without detectable antibodies showed no clinical improvement.

Adolescent↗
PubMed1982

Interferon interactions with thyroid cells.

Iodide uptake by functioning rat thyroid (FRTL) cells is increased by mouse interferon. The effect is detectable using purified interferon; it is not accompanied by an increase in intracellular cyclic AMP levels, is measurable within 20 min, and is prevented by cholera toxin, an agent which inhibits interferon's antiviral activity. The effect of interferon is biphasic with maximally increased iodide uptake (approximately 2-fold) evident at about 300 international mouse units per ml (U/ml) and lesser effects evident at higher concentrations (greater than 1000 U/ml). The effect of mouse interferon on iodide uptake is accompanied by an extremely sensitive antiviral response. Thus, significant antiviral protection is evident at 1 U/ml with FRTL cells, as opposed to 1000 U/ml for nonfunctioning rat thyroid (FRT) cells. Functioning FRTL thyroid cells are also more sensitive to mouse interferon (10-fold) with respect to 2',5'-polyadenylate (An) synthetase activity or to 125I-thyrotropin binding to membrane preparations than are nonfunctioning FRT cells. Antiviral protection in FRTL cells is evident as early as 1 h after exposure to mouse interferon, and is accompanied by a nearly 100-fold increase in the measurable titer of 2'5'-An synthetase activity. Actinomycin D blocks the antiviral effect of interferon, but not its effect on iodide uptake. The results are discussed with respect to the unusually sensitive response of heterologous (rat) cells to mouse interferon: a possible relationship between thyrotropin and interferon receptors; and, the difference in interferon sensitivity exhibited by differentiated (functioning) as opposed to undifferentiated (nonfunctioning) thyroid cells.

2',5'-Oligoadenylate Synthetase↗
PubMed1981

A mouse cell line, which is unprotected by interferon against lytic virus infection, lacks ribonuclease F activity.

A mouse cell line, NIH 3T3, does not respond to some of the activities of interferon. Even after treatment with high concentrations of interferon the replication of lytic viruses, such as encephalomyocarditis virus (EMCV) and vesicular stomatitis virus (VSV) is not inhibited in these cells. In contrast, interferon treatment of these same cells results in the inhibition of Moloney murine leukemia virus (MMuLV) production. We have analyzed enzymatic pathways which are induced by interferon in these cells. After interferon treatment, the level of the (2'-5')oligoadenylate [(2'-5)An] synthetase activity and the phosphorylation of the 67000-dalton protein (P1) are enhanced in NIH 3T3 cells to approximately the same level as interferon-sensitive mouse L-cells. Moreover, NIH 3T3 and L-cells, contain approximately the same levels of enzymes which inactivate (2'-5')An. Both exogenously added (2'-5')A3 or double-stranded RNA (dsRNA) failed to inhibit protein synthesis in NIH 3T3 extracts even though they were potent inhibitors of L-cell extract-directed protein synthesis. Direct measurements of the (2'-5')An-dependent ribonuclease F (RNase F) failed to detect such activity in NIH 3T3 cells. Our results, therefore, suggest that the presence of RNase F activity is necessary for the interferon-induced antiviral activity against EMCV and against VSV. The induction of protein kinase activity by interferon treatment of NIH 3T3 cells appears to have no direct effect on EMCV and VSV replication.

Animals↗
PubMed1980

Spontaneous spatio-temporal organization in yeast cell suspension.

Under suitable conditions of substrate supply, cell suspensions of yeast Saccharomyces uvarum may display synchronized oscillations of glycolytic metabolism. Also in an unstirred suspension, spatial patterns can be induced by continuously feeding enzymically liberated glucose into the system. the absorption of NADH in the suspension was measured in an annular cuvette. The observed structures display moving waves, which travel over a distance of some centimetres in the cell suspension and seem to originate from a volume element in which the phase is ahead of those of its neighbours. The wavelength is estimated to be abot 10 cm and the velocity of spreading of the wave is about 5 cm/min. Waves are only observed in the beginning of glycolytic oscillations. Subsequently the spatial synchrony usually increases and destroys the previous organization. Similar spatio-temporal structures are observed during aggregation of the slime mould Dictyostelium discoideum and in the chemical reaction system of Belousov and Zhabotinsky.

Absorption↗
PubMed1979

Glicentin immunoreactive cells: their relationship to glucagon-producing cells.

The cellular and subcellular localization of one of the gut glucagon-like immunoreactants (GLI-1 or glicentin) and the relative distribution of glicentin- and glucagon-containing cells were investigated by immunocytochemistry. By immunofluorescence, the antiglicentin serum, which does not react with glucagon, revealed positive cells in the islets of Langerhans and in the gut mucosa, particularly in the terminal ileum and colon. In the intestinal mucosa, it was proven ultrastructurally that the glicentin immunoreactive cells correspond to the L cell and that the secretory granules represent the storage compartment of the immunoreactive material. In pancreatic islets, consecutive semithin sections treated with antiglicentin and specific antiglucagon sera showed that the same A cell population reacted with both sera, while immunoperoxidase staining on thin sections revealed that the immunoreactive material was confined to the secretory granules. The same results were obtained on dog oxyntic mucosa, where the glicentin- and glucagon-containing cells were identified as the gastric A cell. The immunocytochemical demonstration of a common glicentin-like material in the A and L cells together with the known presence of a common immunoreactant in glicentin and glucagon strongly support the idea that the A and L cells are ontogenetically related and synthesize their secretory product via a glicentin-like precursor which, by specific cleavage, could yield glucagon and gut glucagon-like immunoreactants.

Animals↗