[Isolation of the Chikungunya virus in Moscow from the Indonesian visitor (September, 2013)].

The results of the virological identification of the Chikungunya fever case in Moscow (September, 2013) in an Indonesian visitor are presented. The clinic, electron microscopy, and molecular genetic data are discussed. The Ghikungunya virus (CHIKV) strain CHIKVILEIV-Moscow/1/2013 belonging to the Asian genotype (ID GenBank KF872195) was deposited into the Russian State Collection of viruses (GKV 1239; 18.11.2013).

[Methods for diagnosis of prion diseases].

The epidemic of bovine spongiform encephalopathy (BSE) in the United Kingdom, its related occurrence of a new type of Creutzfeld-Jacob disease and proven cases of this type of the disease transmitted by blood transfusion initiated intensive studies to develop a inexpensive, prompt, and sensitive method for the early lifetime diagnosis of prion diseases. This would permit initiation of the timely treatment of the patients and prevention of contamination of foodstuffs. However, despite significant progress made in this direction, this objective has not yet been achieved. The present review highlights the currently available methods for the diagnosis of transmissible spongiform encephalopathies, as well as the latest developments in the ultrasensitive detection of these diseases, which is based on the misfolded prion protein complex.

[Detection of hepatitis C virus RNA in the liver biopsy specimens from patients with chronic hepatitis C and hepatocellular carcinoma by means of in situ RT-PCR].

The in situ RT-CPR technique has been first adapted in Russia to detect hepatitis C virus (HCV) RNA in the samples from patients with chronic hepatitis C (CHC) and hepatocellular carcinoma (HC). A total of 18 patients with CHC and HC were examined. Fourteen (78%) samples were ascertained to be positive. A positive reaction was found in 2 of 3 patients with HC. A label was revealed in individual hepatocytes without any regularity in the distribution along the tissue section. A reaction was negative in all samples from control groups. The label was observed in both the cytoplasm and the hepatocytic nuclei. There was no correlation between the degree of hepatic lesion, estimated after Knodelle, the results of the classical RT-CPR and the in situ RT-CPR techniques.

Detection by immunogold techniques of HIV antigens in Lowicryl ultrathin sections of infected cells.

A post-embedding technique for immunocytochemical analysis at the ultrastructural level was used to detect and localize HIV antigens on ultrathin sections of Lowicryl K4M-embedded HIV-infected cells. With serum from an AIDS patient, specific immunogold labelling was obtained exclusively on mature viral extracellular structures. The more intense reactivity was obtained with core antigens. The present immunoelectron microscopy method provides several advantages - high sensitivity of immunodetection, good preservation of cellular morphology, easy preparation procedure - which could lead to the use of this method for HIV-infected human tissues.

Localization by immunogold labelling of HIV-1 structural proteins on Lowicryl embedded HIV-1 infected cell ultrathin sections.

Using several HIV-1-specific antibodies and the immunogold labelling technique, we have detected and localized distinct viral proteins on ultrathin sections of HIV-1 infected cells embedded in the Lowicryl K4M resin. Monoclonal antibodies (MAbs) against p24, p17 and gp160/gp41 showed a preferential labelling on viral formations still attached to the cell membrane (budding process) or free in the extracellular space. The anti-p24 and the anti-p17 MAb yielded a gold labelling not only on mature but also on immature virions where the gold particles were associated with the ring-like electron-dense material. The three HIV-1-specific antibodies against p24, p17 and p55 yielded a cross-reaction with HIV-2 in agreement with the conservation of the internal antigenic determinants of both viruses. In all instances, there was no specific immunogold labelling over the cells, suggesting that once the virus structural proteins were synthesized, they were promptly utilized for virus assembly at the plasma membrane level.

Investigations on the structure and morphogenesis of some arboviruses isolated in the U.S.S.R.

The morphology and some stages of morphogenesis in suckling mouse brains of hitherto serologically unclassified viruses Kaspiy (LEIV-A-63), Zavashan (LEIV-Ap-6158), Artashat (LEIV-A-2366), and Paramushir (LEIV-C-2268), isolated in the U.S.S.R., of GM-710 virus isolated in Scotland, and of Sokuluk (LEIV-K-400) virus belonging to the genus Flavivirus (family Togaviridae) were studied. Virion sizes were determined and changes in infected cells described. Based on their structure and morphogenesis, the viruses Kaspiy, Artashat, Zavashan and Paramushir were referred to the family Bunyaviridae and GM-710 virus was referred to the genus Orbivirus (family Reoviridae).

Razdan virus, a new ungrouped bunyavirus isolated from Dermacentor marginatus ticks in Armenia.

A virus, designated Razdan, was isolated from Dermacentor marginatus ticks in the Armenian S.S.R. in 1973. The complement fixation tests revealed no antigenic relationships to 74 tick-borne arboviruses. The size of the virus is about 100 nm; it agglutinates goose erythrocytes at pH 5.5--7.0, is pathogenic for newborn, 14-day-old and adult white mice and multiplies in primary and continuous cell cultures. Morphological properties of the virus permit its classification as a member of the family Bunyaviridae.

Persistent SV5 virus infection in continuous cell cultures.

A continuous line of guinea pig kidney cells (CGPK/H) and a continuous line of mouse fibroblasts (L/H) spontaneously infected with parainfluenza virus SV5 were found. These cultures showed no enhanced cell degeneration or symplast formation, nor was haemagglutinin accumulation or infectious virus demonstrated in them. However, regular reproduction of ribonucleoprotein (RNP) characteristic of parainfluenza viruses, morphologically complete virions and antigens producing antibody to SV5 virus were found in the cells. Focal haemadsorption neutralized by antiserum to SV5 virus was also demonstrated. The infection persisted in the cell populations for over 2 years (the observation period) under standard conditions of cell dispersion and subcultivation.

Sikhote-Alin virus, a new member of the cardiovirus group (Picornaviridae) isolated from Ixodes persulcatus ticks in Primorie Region.

A virus, designated Sikhote-Alin, was isolated in 1970 from Ixodes persulcatus ticks collected from a wild boar in the Primorie region (U.S.S.R.) Sikhote-Alin virus showed no haemagglutinating activity and no antigenic relationships with arboviruses of 12 antigenic groups, 17 ungrouped tick-borne arboviruses, porcine enteroviruses and coxsackie A (types 1-18) viruses. An one-way antigenic relationship was demonstrated by complement fixation with cardioviruses (Mengo and Columbia-SK strains). The virus contains RNA, is resistant to lipid solvents, highly thermostable in the presence of 1 M MgCl2 and its size is over 20 nm but less than 25 nm. All these properties make it possible to consider it as a new member of the cardiovirus group (genus Enterovirus; Picornaviridae).

Khasan virus, a new ungrouped bunyavirus isolated from Haemaphysalis longicornis ticks in the Primorie region.

Strain LEIV-776P, possessing no haemagglutinating activity was isolated in 1971 in the Primorie region (U.S.S.R.) from Haemaphysalis longicornis Neumann 1091 tick. Complement fixation (CF) tests revealed no antigenic relationship with 24 antigenic groups of arboviruses or 21 ungrouped viruses isolated from ticks. The virus isolate contains RNA and is relatively sensitive to ether and sodium deoxycholate; it is pathogenic for suckling mice and two-week-old white mice by the intracerebral route, and replicates in primary cultures of chick and duck fibroblasts and green monkey kidneys and in a continuous line of pig embryo kidney cells without any cytopathic effect. According to electron microscopy, its size is 90-110 nm. Its morphological properties permit to classify it as a member of the family Bunyaviridae.

[Structure and regulation of the assembly of bacteriophage T4 fibrillar proteins. II. Isolation and initial structural characteristics of whiskers]. / Struktura i reguliatsiia sborki fibrilliarnykh belkov bakteriofaga T4. II. Vydelenie i nachal'naia strukturnaia kharakteristika vorotnichkovykh nitei.

A procedure for purification of bacteriophage T4 whiskers and it's monomeric subunits--gene product wac--has been developed. We have shown, that the whiskers are composed of two identical copies of gene product wac with molecular weight of 56 kDa each. The dimer of gene product wac is a highly ordered structure and it's length is about 70.0 +/- 10.0 nm, as revealed by electron microscopy. The amino acid composition of whiskers is very similar to that of watersoluble keratins. We have proposed a new term for the definition of the whiskers--the fibritin.

[Effectiveness of incorporation of spermine-condensed viral DNA into liposomes. Interaction of liposomes with cells]. / Effektivnost' vkliucheniia kondensirovannykh sperminon virusnykh DNK v liposomy. Vzaimodeistvie liposom s kletkami.

DNA from bacteriophage lambda or monkey adenovirus type 7 have been condensed with spermine and included into three types of liposomes: ethereal, obtained by inverted phases technique and by Ca++ fusion. Compact DNA form presents a tor with 100 nm external diameter and 430 nm width. It can be included into 100 nm or larger liposomes. Total preparation contains 15-18% of the required liposomes. Liposomal fractions with included DNA were separated from empty liposomes by step gradient of ficoll 400. Liposomal fraction having included DNA contains 15-18% of common lipid. Liposomal interaction with monkey cell cultures has been studied.

[Morphogenesis of Okhotsky virus from the genus orbisvirus]. / Morfogenez virusa Okhotskii iz roda orbisvirusov

Morphogenesis of Okhotsky virus isolated in the USSR was studied. Virions of 50-65 nm in diameter were found in the peripheral zone of cytoplasmic matrices. Crystalline-like arrays of virions were observed not reported by other investigators studying morphogenesis of orbisviruses. Besides, not directly associated with matrices, crystalline arrays of some proteins were found in infected cells for the first time with orbisviruses. Nevertheless the main stages of morphogenesis of Okhotsky virus were similar to those of other members of the orbisvirus group.

[The localization of delta antigen in the hepatocytes in chronic delta hepatitis]. / Lokalizatsiia del'ta-antigena v gepatotsitakh pri khronicheskom gepatite del'ta.

The results of immunohistochemical and electron microscopic examinations of liver biopsies from 47 patients with chronic delta hepatitis are presented. Delta antigen (HDAg) was detected by immunoperoxidase method in the liver of 35 (74.4%) patients. HDAg was demonstrated in hepatocyte nucleus alone in 22 (62.9%) patients, simultaneously in the nucleus and the cytoplasm in 9 (25.7%), in the cytoplasm alone in 4 (11.4%). In nuclear localization, delta antigen may fill the entire nucleoplasm or only the peripheral zone of the nucleus. When delta antigen fills the entire nucleus, hepatocytes undergo more marked pathological changes than those in which HDAg is localized in the nuclear periphery or those where no delta antigen was found.

[Possibility of detecting the matrix protein of the influenza virus in intact and disrupted virions by immunoenzyme analysis and immune electron microscopy]. / Issledovanie vozmozhnosti vyiavleniia matriksnogo belka virusa grippa v intaktnykh i razrushennykh virionakh s ispol'zovaniem immunofermentnogo analiza i immunnoi élektronnoi mikroskopii.

ELISA and immune electron microscopy were used to study possible causes of the detection of antigenic reactivity of influenza virus matrix protein in purified virus suspension directly adsorbed on polystyrene. No interaction of antibody to M protein with the surface of virus and subvirus particles was observed. The process of virus sorption on polystyrene for a long period was shown not to lead to disruption of intact virus particles, and the detection of the internal matrix protein in preparations of purified influenza virus was due only to the presence of partially or completely destroyed virions in the viral suspension.

[Isolation and characteristics of endocytic vacuoles containing the influenza virus]. / Vydelenie i kharakteristika éndotsitarnykh vakuolei, soderzhashchikh virus grippa.

Endocytic vacuoles (receptosomes) containing influenza virus were isolated from the cytoplasm of Ehrlich ascitic carcinoma cells and characterized. In the sucrose density gradient, the virus-containing material was detected in two peaks with a buoyant density of 1.175-1.16 and 1.155-1.135 g/cm3 with which the activity of marker enzymes of cell plasma membranes was associated. The virus was present in receptosomes in morphologically and electrophoretically intact condition. Examinations for the lipid composition of endocytic vacuoles showed the presence in their membranes of large amounts of cholesterol and glycolipids, particularly asialo-GM1 which, according to some authors may enhance the fusion of viral and cell membranes.

[Isolation of polymeric glycoprotein complexes of enveloped viruses using the Quill A glycoside and a study of their immunogenic activity]. / Poluchenie mul'timernykh kompleksov glikoproteidov obolochechnykh virusov s glikozidom Kvil A i izuchenie ikh immunogennoi aktivnosti.

Specific multimer complexes of glycoproteins of enveloped viruses were prepared by treatment of suspensions of purified concentrated virus with a non-ionic detergent MECK mixed with 0.2% glycoside Quil A. Mixed complexes of glycoproteins with glycoside Quil A were formed upon removal of the detergent from the mixture of solubilized glycoproteins and glycoside Quil A. According to the results of electron microscopy, the formed complexes differed morphologically from the conventional micelles of glycoproteins and presented spherical reticular structures 20-30 nm in diameter. The immunogenic potency of the complexes was much higher than that of virus particles and micelles of purified glycoproteins and was comparable to the immunogenic potency of glycoproteins mixed with complete Freund adjuvant. The protective activity of the complex of protein G of rabies virus with glycoside Quil A was higher than that of the subunit rabies vaccine adsorbed on aluminium hydroxide.

[Expression of human immunodeficiency virus gag antigens and formation of virus-like particles in a cell culture, infected with recombinant vaccinia virus strains (an electron-microscopic study)]. / Ekspressiia gag-antigenov virusa immunodefitsita cheloveka i obrazovanie virusopodobnykh chastits v kul'ture kletok, infitsirovannykh rekombinantnymi shtammami virusa ospovaktsiny (élektronno-mikroskopicheskoe issldovanie).

The synthesis of gag antigens of human immunodeficiency virus (HIV) by recombinant vaccinia virus strains containing the expressed genes gag and gag-pol and the capacity of these proteins to formation of virus-like particles during infection of various cell cultures were studied. The recombinant strain containing the truncated gag gene (p48gag) was shown to effectively synthesize gag polypeptides and to form immature virus-like particles during infection of all the cell cultures tested (CV-1, Hep-2, HT-29). The morphogenesis of mature virus-like particles was detected by electron microscopy only in infection of Hep-2 cells with a strain containing a complete gag-pol sequence of HIV.

[Structural study of influenza A virus virions using cryogenic methods]. / Issledovanie struktury virionov virusa grippa A kriogennymi metodami.

The method of cryospalling was used to study formation of influenza A virus virion on the plasma membrane and to study the inner topography of virus membrane. This method is significantly superior to the others in that cryofixation of the native material constitutes one of its steps. The experiments showed that all budding virus particles were characteristically elongated and virions separating from the membrane acquired spherical shape. The mechanism of this phenomenon is discussed. Characteristics of four types of spalls of isolated virions are presented. An assumption is made with regard to the nature of intramembrane particles on the E-plane of influenza virion lipid membrane.

[Electron microscopic study of genome homology in 2 clones of the simian adenoviruses type 7]. / Elektronno-mikroskopicheskoe izuchenie gomologii genomov dvukh klonov adenovirusov obez'ian tipa 7.

Homology of the genomes of large-plaque (lp) and small-plaque (sp) clones of simian adenovirus type 7 (SV7) was studied experimentally. Guanidine chloride helped to isolate circular DNA molecules of identical length for both clones within the measurement error (1 = 11 +/- 0.2 micrometer). A heteroduplex SV7lp--SV7sp molecule was found to have two nonhomology sites 0.5 +/- 0.3 mu and 1.0 +/- 0.5 micrometer long located at a distance of 2 and 4 micrometer from one of the ends of the molecules, respectively.