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Biomedical subjects

R R Dourmashkin

Publications and source records attributed to R R Dourmashkin.

16 recordsLinked to original sources

Studies on the mechanism of influenza virus entry into cells.

Inhibitors of glycolysis, oxidative phosphorylation, protein synthesis, membrane Na&-K& transport and microfilament and microtubule function have been employed to elucidate the mechanism of influenza virus uptake by CAM and CEF cells. Electron microscopy demonstrated uptake of virus by viropexis in the presence of all these inhibitors. Utilizing a pulse labelling technique, virus entering CEF cells in the presence of inhibitors was shown to initiate specific virus polypeptide synthesis after neutralization of remaining extracellular virus and removal of the inhibitors. As a consequence of these findings an energy independent mechanism of viropexis has been proposed.

Animals

Interaction of Mycoplasma pneumoniae with human lung fibroblasts: characterization of the in vitro model.

The interaction of pathogenic Mycoplasma pneumoniae and host cells was studied in cell cultures of MRC-5 human lung fibroblasts. A comparison of results obtained with fibroblasts in a monolayer format and with hamster tracheal explant cultures indicated that the former can bind significantly larger numbers of mycoplasmas. In addition, the attachment was 96% specific, that is, mediated through a neuraminidase-sensitive receptor on the host cell. Uptake of mycoplasmas was directly related to the number of mycoplasma cells present in the inoculum, and attachment was virtually complete within a 30-min period at 37 degrees C. High doses of M. pneumoniae induced a marked cytopathic effect, whereas doses of less than or equal to 10(6) colony-forming units per ml produced grossly observable cell damage that was moderate and variable. Transmission electron microscopy studies indicated that attachment of M. pneumoniae to the surface of lung fibroblasts occurred with the specialized terminal structure or binding site oriented closest to the epithelial cell surface. The filamentous mycoplasma cells were spatially arranged in several configurations and were not limited to a vertical orientation. The advantages and disadvantages of human lung fibroblast monolayer cultures, in reference to other in vitro models are discussed. A new mycoplasma agar medium (G-200 agar) with a defined tissue culture base and 10% horse serum is also described.

Binding Sites

A year's experience of the rotavirus syndrome and its association with respiratory illness.

In a hospital study rotavirus was identified in 51% of 152 children with diarrhoea. These patients showed a clinical pattern that was distinct from patients in whom the diarrhoea was associated with bacteria, other viruses, or no pathogens. A respiratory illness was described in 66% of rotavirus patients and usually preceded the gastrointestinal symptoms. Vomiting lasted between one and 3 days and was curtailed by substituting the normal diet with clear fluids. Watery diarrhoes continued for 4 or 5 days, even when rehydration was by the intravenous rather than the oral route. Prolonged diarrhoea was rare. Most children infected with rotavirus were under 2 years of age, but dehydration was most severe in infants aged between 12 and 18 months. A clinician can thus recognise the rotavirus syndrome and expect spontaneous recovery if adequate rehydration is maintained for a critical few days.

Adolescent

Growth and effect of chlamydiae in human and bovine oviduct organ cultures.

Organ cultures of 10 Fallopian tubes were inoculated with a genital strain of Chlamydia trachomatis and seven were infected. Infection was enhanced by centrifuging the organisms on to the tissues, larger numbers of organisms being reisolated from the tissues after this procedure. There was evidence of chlamydial multiplication because the number of organisms which were recovered from the tissues three to five days after inoculation had increased. Recovery was rare, however, after the sixth day, thus suggesting a self-limiting infection. Organ cultures of two bovine oviducts were infected with the bovine abortion strain of Chlamydia psittaci, but in these experiments centrifugation of the inocula did not enhance infection. The organisms were found in both the tissue and medium of cultures up to 18 days after inoculation and in much greater numbers than in the C. trachomatis-infected Fallopian cultures. Chlamydial infection was not entirely host-tissue specific, because C. trachomatis organisms were isolated from bovine oviduct cultures. Inclusions, however, were not detected histologically or electron microscopically in the epithelium of C. trachomatis-infected cultures, but they were detected by these means in C. psittaci-infected bovine cultures. All the elements of the chlamydial growth cycle were seen by electron microscopy, organisms being found in ciliated and possibly non-ciliated cells, and shedding of some infected epithelial cells was observed. No evidence of extensive epithelial cell damage was observed, however, and no loss of ciliary activity was detected in cultures infected with either C. trachomatis or C. psittaci when compared with uninoculated cultures. Thus acute salpingitis, when caused by chlamydial infection, is probably immunologically mediated.

Animals

The structural events associated with the attachment of complement components to cell membranes in reactive lysis.

Electron microscopic study of the events occurring at the cell membrane during reactive lysis by complement, showed that a foliaceous particle was formed at the C5b-7 stage, that enlarged to a particle with a variable number of arms at the C5b-8 stage. Up to this point, no typical complement lesions were found. At the C5b-9 stages, the particles were completely converted to typical complement lesions, i.e. hollow cylinders projecting from the cell membrane and partly penetrating it. C5b-9 complexes assembled in the fluid phase did not show the typical structure of the lesions, but were amorphous masses of fibres.

Complement C5

Relationship of pili to colonial morphology among pathogenic and nonpathogenic species of Neisseria.

Growth in colonies with type 1 morphology and the presence of pili are characteristics that have been associated with virulence of gonococci for humans. To determine whether the presence of pili per se might be responsible for colony type 1 morphology, the relationship of pili to colony type was examined in various species of Neisseria. Short pili (175 to 210 nm in length) were seen only on nonpathogenic neisseria, whereas long pili (up to 4,300 nm) were seen on organisms of both nonpathogenic and pathogenic species. Although long pili, similar to those found on organisms from high-domed, type 1 colonies of gonococci, were observed on organisms from high-domed, type 1 colonies of nonpathogenic Neisseria species, they were also observed on low-convex, type 4 colonies of meningococci and nonpathogenic neisseria. Among meningococci there was no difference in the morphology of colonies consisting of organisms with many long pili and colonies consisting of organisms that completely lacked pili. Thus, there was no consistent relationship of pili to colonial morphology. Unless the pili of N. gonorrhoeae are unique among Neisseria species in their influence on colonial morphology, it is likely that factors other than pili determine colony type 1 morphology of gonococci. Whether these same factors, either alone or in conjunction with pili, are also responsible for gonococcal virulence warrants further investigation.

Neisseria

Polypeptides of the surface projections and the ribonucleoprotein of avian infectious bronchitis virus.

Purified avian infectious bronchitis virus was digested with bromelain (0.7 mg/ml), and the surface projections were removed. Polyacrylamide gel electrophoresis of the polypeptides from these bromelain-treated particles showed that VP1, VP2, and VP5 were missing from the seven polypeptides. VP1 to VP7, that were present in untreated virus preparations. Milder bromelain treatment (0.07 mg/ml) left visible surface projections and polypeptides comprising VP1 and VP2 intact, but removed VP5. Thus, there are apparently two types of surface projections on the virus particle. The ribonucleoprotein complex was released from virus particles disrupted with 1% Nonidet P-40. The proportion of VP6 in such preparations was greatly reduced, implying that VP6 is the structural polypeptide of the ribonucleoprotein. Polypeptides VP1, VP2, VP4, and VP5 are glycosylated, but none of the polypeptides contains lipid.

Bromelains

Lymphocytic infiltration of the brain in sleeping sickness.

Cerebrospinal fluid mononuclear cells from 40 patients with advanced Gambian sleeping sickness were examined for intracytoplasmic immunoglobulin and for B- and T-lymphocyte markers. About 5% of mononuclear cells were plasma cells. Most of the lymphocytes present were B cells. These findings suggest that the considerable lymphocytic infiltration of the nervous system seen in advanced sleeping sickness is not a cell-mediated immune reaction to trypanosomes. Immune complexes may play a part in producing the brain damage characteristic of this disease.

B-Lymphocytes

Nonpilar surface appendages of colony type 1 and colony type 4 gonococci.

The nature and distribution of surface appendages morphologically distinct from pili were examined in three strains of gonococci. As reported previously by others, large appendages were seen emanating from colony type 1 gonococci when negatively stained preparations were examined by electron microscopy. Whereas the diameter of pili was 4 to 6 nm, the appendages varied from 8 to 40 nm in diameter with bulbous enlargements of up to 130 nm. However, in contrast to previous findings, the appendages were not limited to virulence-associated colonial types but occurred with almost equal frequency on organisms from colonial types 1 and 4. Thus, the presence of these appendages on gonococci was unrelated to colonial morphology, and there is currently no reason to associate them with gonococcal virulence. The large appendages were seldom seen if the organisms were suspended in isotonic fluid rather than in distilled water, which was used in previous studies. This suggests that the large appendages may occur infrequently in the isotonic environment of tissues or in slightly hypertonic purulent exudates. Nevertheless, we did observe small appendages or cell wall blebs on the surface of gonococci suspended in isotonic medium and noted similar structures as spheres free in the surrounding milieu. These findings suggest that the material of which the large appendages consist may form small appendages or cell wall blebs and be shed into the milieu under isotonic conditions. Such cell wall blebs have been demonstrated on meningococci and been shown to consist of endotoxin.

Cell Wall