Antiviral activity of liposomal preparations of antibiotic geliomycin.

A liposomal preparation with maximally possible content of incorporated geliomycin was obtained. Its cytotoxicity was studied in a culture of human embryonal diploid fibroblasts. Antiviral activity was studied on a model of cytomegaloviral infection in vitro by the capacity to plaque formation. Liposomal geliomycin was 10-fold less toxic for human cells than the solution of the antibiotic in DMSO and exhibited antiviral activity towards cytomegaloviral infection at a concentration of 0.042 microg/ml.

[Detection of direct markers of cytomegalovirus. Research of the spectrum and avidity of antiviral antibodies in infants and preschool children].

Thirty-three children aged 1 month to 3 years were examined within the case study. spELISA, immunoblot (IB), shell vial method (SVM) and PCR, were used for the detection of anti-CMV IgM and IgG, in the diagnosis of cytomegalovirus (CMV). Clinical signs of CMV infection (CMVI) were registered in 20 children (group 1); no CMVI specific signs were detected in the remaining 13 children (group 2). Class M antibodies were identified in 50% of group-1 sera. Around 80% of children in the group had anti-CMV-IgG. AI < 0.6 was in 3 (20%) of 15 examinees. Direct CMV markers (DNA and infection activity) were detected in 13 (65%) of 20 children. Sera of 13 children with non-specific symptomatology (group 2) had no anti-CNV-IgM, while IgG were found in 54% examinees in the group. The infectious active virus was not detected in a single baby. The used laboratory tools enhance the efficiency of CMVI diagnosis and denote a disease variation.

[Activation of transcription of ribosome genes following human embryo fibroblast infection with cytomegalovirus in vitro]. / Aktivatsiia transkriptsii ribosomnykh genov pri tsitomegalovirusnoi infektsii fibroblastov émbriona cheloveka in vitro.

Cytomegalovirus (CMV) infection of human diploid embryo fibroblasts in vitro causes a massive cell death on the 3-4th day of infection with a high primary infection coefficient (1-5 U/cell). Cytopathological effects of viral infection on the 3-4th day includes diminishes of the cell size, changes in their form, compaction of the nuclear chromatin, and disorganization and inactivation of the nucleolus. However, the early stages of the viral infection progression (the 1st-2nd day) are accompanied by unequivocal activation of rDNA (ribosomal gene) and the bulk of chromatin transcription. There are several features to support this conclusion: In the early CMV-infected cell 1) the nucleolar size is increased; 2) the number of intranucleolar foci binding the specific RNA-polymerase I transcription factor (UBF) is augmented; 3) the Ag-NOR staining is enhanced; 4) 3H-uridine incorporation to the nucleoli is activated; 5) the ultrastructure of the nucleolus is changed. Altogether, these data argue in favor of activation of rDNA transcription in human fibroblasts in vitro at the initial stages of infection.

[Comparative efficacy of laboratory methods for cytomegalovirus detection in autopsy material]. / Sravnenie éffektivnosti laboratornykh metodov vyiavleniia tsitomegalovirusa v autopsiinom materiale.

Eighty eight autopsy specimens obtained from 30 fetuses, still-borns and infants died during the first year of life, all suspected for congenital virus infection at postmortem examination, were studied. The specimens were analyzed by 3 techniques: rapid culture method (RCM) for detection of cytomegalovirus (CMV) infectious activity, the immunocytochemical method for detection of CMV antigen in prints of organs and polymerase chain reaction (PCR) for detection CMV DNA. CMV was detected in 16 out of 26 specimens (61.5%) by PCR, in 43 out of 88 specimens (49%) by RCM and in 15 out of 64 specimens (23%) in prints. The comparison of immune reagents revealed that monoclonal antibodies (McAb) were more specific than polyclonal serum antibodies, as the latter yielded the positive reaction in 10 out of 26 cases (38%), found to be negative in PCR. The data thus obtained indicate that complex techniques, including PCR and RCM in combination with McAb, should be used for evaluation of CMV infection role in child mortality.

[Suppression of cytomegalovirus infection in cell system by fullerene amino acid derivates]. / Podavlenie tsitomegalovirusnoi infektsii v kletochnoi sisteme aminokislotnymi proizvodnymi fullerena.

Effects of two water-soluble derivatives of fullerene C60-o-aminocaproic acid (C60-ACA) and C60 sodium salt of omega-aminocaproic acid (C60-Na-ACA) on in vitro cytomegalovirus (CMV) infection were studied. C60-Na-ACA 4-5-fold inhibited the cytopathic effect of CMV in comparison with C60-ACA, the effective dose for C60-Na-ACA being 0.6 microgram/ml and that for C60-ACA 2.7 micrograms/ml. Immunocytochemical analysis of virus proteins in infected cells has shown that C60-Na-ACA inhibits the production of late structural CMV protein gB, but does not modify the expression of immediate early nonstructural protein IEp72. Studies of cell viability, growth characteristics, and DNA synthesis revealed that the cytotoxic effect of C60-Na-ACA on human diploid fibroblasts in negligible, the cytotoxicity index varying from 160 to 1500 micrograms/ml in different tests. Selectivity index for C60-Na-ACA is 267-2500, which differs negligibly from that of gancyclovir (100-1000), while the cytotoxicity of C60-Na-ACA is essentially lower.

[Effect of cytomegalovirus on cell cycle progression and formation of pathological mitoses in cultured human diploid fibroblasts]. / Vliianie tsitomegalovirusa na prokhozhdenie kletochnogo tsikla i formorovanie patologicheskikh mitozov v kul'ture diploidnykh fibroblastov cheloveka.

The effect of cytomegalovirus on the cell cycle was studied autoradiographically in an asynchronous culture of human diploid fibroblasts. The analysis of labeled mitosis showed that some cells infected in the S phase ceased to progress through the cell cycle at one of its phases (S, G2, or M); at the same time, at least part of infected cells remained capable of entering mitosis. Beginning from day 2 after infection by cytomegalovirus, the accumulation of pathological mitotic cells blocked at metaphase was observed in the culture. Approximately 50% of these cells contained 3H-thymidine label above chromosomes. This fact suggested the possibility of pathological mitosis in cells that were infected both at the S and other phases of the cell cycle. The detailed morphological analysis of chromosomes at different stages of infection demonstrated that the degree of their morphological changes increases from slight (stronger condensation) to severe pathology (fragmentation). In the aggregate, the results of the study suggested that abnormal chromosome morphology resulted from irreversible cell division arrest under the effect of cytomegalovirus.