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Biomedical subjects

G Kaluza

Publications and source records attributed to G Kaluza.

34 records · Page 2Linked to original sources
PubMed1981

Enhancement of plaque formation and cell fusion of an enteropathogenic coronavirus by trypsin treatment.

Plaque formation, replication, and related cytopathic functions of the enteropathogenic bovine coronavirus strain L9 in bovine fetal thyroid (BFTy) and bovine fetal brain (BFB) cells were investigated in the presence and absence of trypsin. Plaque formation was enhanced in both cell types. Plaques reached a size with an average diameter of 5 mm within 4 days with trypsin in the overlay, whereas their diameter remained less than 1 mm at this time after plating without trypsin in the overlay. Fusion of both cell types was observed 12 to 18 h after infection when trypsin was present in the medium. Fusion was not observed in infected BFB cell cultures and was rarely observed 48 h after infection of BFTy cells maintained with the trypsin-free medium. The largest polycaryons formed had 15 to 22 nuclei. They then lysed and detached. Cell fusion depended on de novo synthesis of hemagglutinin and infectivity. Fusion from without was not observed. Virus produced under trypsin-enhancing conditions accompanied by cell fusion did not lyse mouse erythrocytes that reacted with L9 coronavirus hemagglutinin. Trypsin-treated, infected BFTy cultures produced coronaviral particles that excluded stain from the envelope confinement. These virions had uniformly shorter surface projections than did the viral forms generated by trypsin-free cell cultures.

Animals↗
PubMed1979

Semliki forest virus: cause of a fatal case of human encephalitis.

A fatal case of human encephalitis has been observed for which our results indicate that Semliki Forest virus (SFV) was the etiologic agent. This is surprising in view of the fact that this virus, which has been widely studied, was believed to be one of the arboviruses nonpathogenic for man. Described are the clinical course, the virological examinations performed, and the histopathological findings in the central nervous system.

Adult↗
PubMed1976

Early synthesis of Semliki Forest virus-specific proteins in infected chicken cells.

Cells preinfected with fowl plague virus followed by treatment with actinomycin D are a suitable system for studying early protein synthesis in cells infected with Semliki forest virus. One and one-half hours after superinfection, three new nonstructural proteins (NVP) were detected: NVP 145, NVP, 112, and NVP 65. They appeared in parallel with a low incorporation of mannose at the beginning of the infectious cycle. Behavior on chasing suggested a precursor relationship of NVP 112 to the envelope glycoproteins. Two kinds of NVP 65 are described, both of which are varieties of NVP 68 with an incomplete mannose content. One type, detected early after infection, was converted into NVP 68 by supplementary glycosylation. The second, late type was stable. It contains fucose and resembles the NVP 65 observed after impairment of glycosylation. The mechanism of NVP 68 glycosylation is discussed. The presence of the complete carbohydrate moiety is crucial for the cleavage of NVP 68 into the envelope proteins E2 and E3 and, thus, for virus maturation. Only the complete form of NVP 68 was precipitated by envelope-specific antisera. A large production of NVP 78 is a further feature of the early events in infected cells. It is not related to the structural proteins.

Animals↗
PubMed1975

Effect of impaired glycosylation on the biosynthesis of Semliki forest virus glycoproteins.

The glycoproteins of Semliki Forest virus, grown in chicken embryo cells, were labeled with radioactive sugars. The data indicate a high mannose content of the nonstructural precursor glycoprotein NSP 63. This protein can also be readily labeled with 2-deoxy-D-glucose. The envelope glycoproteins E1 and E2 are relatively rich in galactose, glucosamine, and fucose. Glycosylation can be impaired by 2-deoxy-D-glucose or D-glucosamine or by omission of sugars in the culture medium. Under these conditions characteristic changes in the electrophoretic profile of the viral polypeptides are observed: in the regions of glycoproteins NSP 97, NSP 63, and E1 and E2 new protein peaks can be detected. These polypeptides seem to be aberrant forms of the glycoproteins. When compared with the normal molecules they have lower molecular weights and contain less carbohydrates, especially mannose. Pulse-chase experiments indicate that the altered glycoproteins are degraded very slowly if at all. If, however, impairment is caused by omission of sugars in the culture medium, the radioactivity is chased after addition of glucose from the region between NSP 63 and E1 + E2 into the E1 + E2 peak. This suggests a completion of the carbohydrate chains under these conditions.

Cells, Cultured↗
PubMed1975

Inhibition of cellular protein synthesis by simultaneous pretreatment of host cells with fowl plague virus and actinomycin D: a method for studying early protein synthesis of several RNA viruses.

A method is described for analysis of viral protein synthesis early after infection when minute amounts of viral proteins are effectively concealed by large amounts of produced host-specific proteins. The method is superior to a radioimmune assay, since all virus-induced proteins can be measured independent of their immunological reactivity. Host-specific protein synthesis can be suppressed by infection with fowl plague virus. Addition of actinomycin C 1.25 h postinfection does not prevent this suppression, but it does block effectively the formation of fowl plague virus-specific proteins. Such cells synthesize only small amounts of cellular proteins, as revealed by polyacrylamide electrophoresis. They can be superinfected with several different enveloped viruses, however, without significant diminution of virus yeilds. In pretreated cells the eclipse is shortened for Semliki Forest virus, Sindbis virus, and vesicular stomatitis virus, but prolonged for Newcastle disease virus. The onset of protein synthesis, specific for the superinfecting virus, could be clearly demonstrated within 1 h after superinfection. At this time, in cells superinfected with Semliki Forest virus, great amounts of NSP 75 (nonstructural protein; molecular weight, 75 X 10(3)) and reduced amounts of the core protein C could be deomonstrated. The precursor glycoprotein NSP 68 is followed by a new polypeptide, NSP 65: three proteins with molecular weights exceeding 100 X 10(3) were observed which are missing later in the infectious cycle. Similar results were obtained after superinfection with Sindbis virus. The formation of a new polypeptide with a molecular weight of about 80 X 10(3) was detected. After superinfection with vesicular stomatis virus or Newcastle disease virus the formation of new proteins, characteristic for the early stage of infeciton, was not observed.

Cell Line↗
PubMed1975

Interference with the glycosylation of Semliki Forest virus proteins.

It has been shown that under defined conditions the multiplication of SFV can be inhibited by specific interference with the production of viral glycoproteins. These conditions are: a) Either 0.2 mM dGlc or 2 mM glucosamine in glucose-containing medium, b) lack of any sugar or energy source in the culture medium, c) pyruvate as energy source, late in the infectious cycle. Under these conditions underglycosylated glycoproteins occur, which cannot be used to form physical or infectious particles. It is not yet clear whether the appearance of a heterogeneous population of molecules in the molecular weight range between 50 and 63,000 is due exclusively to different lengths of the carbohydrate side chains, or also due to wrong splitting or partial degradation of the precursor molecules by cellular proteases. Glucosamine seems to act via the accumulation of UDP-N-acetylglucosamine (or a derivative of it), while dGlc is metabolized along the pathway of mannose and acts mainly as an antimetabolite of mannose.

Culture Media↗
PubMed1974

Inhibition of the multiplication of vesicular stomatitis and Newcastle disease virus by 2-deoxy-d-glucose.

The production of infectious vesicular stomatitis (VSV) and Newcastle disease virus can be completely inhibited by 2-deoxy-d-glucose in pyruvate-containing medium, if virus either grown in pyruvate-containing medium or dialyzed against phosphate-buffered saline is used for infection. Under these conditions, the synthesis of all VSV proteins is reduced. VSV RNA, which is synthesized at reduced rates, seems to be unstable. The effect is completely reversible. If virus grown in glucose-containing medium is used for infection, the production of both viruses is not significantly inhibited by 2-deoxy-d-glucose. Under these conditions the production of the VSV glycoprotein is specifically impaired, but does not lead to a marked reduction of the yield of infectious virus.

Amino Acids↗