[Diagnosis and importance of the opportunistic herpes-virus infection in etiology and pathogenesis of different eye diseases]. / Gerpesvirusnaia infektsiia v étiopatogeneze razlichnykh zabolevanii glaz.

A total of 439 patients (adults--184, children--255) with herpetic keratitis as well as with uveitis and choriorenitis of different geneses and localizations were examined. An aggravation of HSV-infection was determined by detecting, in the blood), anti-bodies to the unstructured early viral antigens by using the immune enzyme analysis (IEA). The efficiency of the test was demonstrated during the examination of patients with different ophthalmic pathologies. An active infection was detected in 16-39% of patients with various ocular diseases of non-herpetic etiology. The pathogenetic value of reactivated HSV infection is diverse: it can function as an etiological or trigger factor and it can aggravate the main disease through causing, postoperatively, complications.

[Virus-like particles as a source for obtaining antigens for diagnosing rubella]. / Virusopodobnye chastitsy kak istochnikh polucheniia antigenov dlia diagnostiki krasnukhi.

Rubella diagnostic agents for hemagglutination inhibition (HI) and enzyme immunoassay (EIA) based on rubella virus-like particles (RVLP) have been developed. Noninfectious RVLPs containing three structural E1, E2, and C proteins were expressed in transfected CHO24S cell culture. HI titer in culture medium was 1:256. Tween-80 treatment and ether increased HI titer 4-6-fold and rendered the antigen higher stability. Immunogenic properties of RVLPs were similar to the native rubella virus in HI test with international reference human rubella serum and sera from convalescents after rubella. Serum of mice immunized with RVLP reacted similarly with RVLP antigens and native virus. Antigen for EIA from RVLP was prepared by concentrating RVLP from culture fluid and partial purification by ultracentrifugation. The results of human sera testing by HI and EIA with RVLP and native virus antigens coincided. RVLP are identical to antigenic structure of the virus, are stable and easily purified, and can therefore be used for commercial production of HI and EIA antigens.

[Prevalence and clinical significance of active cytomegalovirus infection in patients with inflammatory ophthalmopathy]. / Rasprostranennost' i klinicheskoe znachenie aktivnoi tsitomegalovirusnoi infektsii u bol'nykh s oftal'mopatologiei vospalitel'nogo kharaktera.

A total of 374 patients with different inflammatory diseases of the eyes were examined (227 children and 147 adults). For detecting active cytomegalovirus (CMV) infection, sera were tested for antibodies to nonstructural intact CMV proteins. Antibodies were detected in 26 of 147 adults (17.7%) and 15 of 227 children (6.6%). Difference in the frequency of detection of active CMV infection in adults and children is statistically significant (p < 0.01). Three groups of patients with active CMV were distinguished: children with congenital diseases, immunocompromised patients, and the most numerous group of patients with isolated ocular diseases without manifest signs of immunodepression. In this latter group active CMV was detected in patients with different clinical diagnoses, including ophthalmic herpes. Russian commercial enzyme immunoassay kit CMV-Control proved to be a highly sensitive, simple, and cheap test for the diagnosis of exacerbations of chronic CMV infection.

[Cytomegalovirus infection in children with endogenous uveitis]. / Tsitomegalovirusnaia infektsiia u detei s éndogennymi uveitami.

A total of 405 children aged 3 months to 15 years with uveitis of different origin and localization and 50 mothers of children with intrauterine uveitis were tested for cytomegaloviruses (CMV). Chronic CMV infection was detected in 79% children and 88% mothers. Active CMV infection was diagnosed in 7.1% children with various clinical forms of uveitis; it was somewhat more frequent in cases with grave posterior uveitis and panuveitis complicated by detachment of the retina and vitreous fibrosis. Anti-CMV IgG antibodies indicate an infection, but their detection is insufficient for identifying the etiology of uveitis. Active CMV infection can be a cause of uveitis or aggravate uveitis of another etiology and favor the development of postoperative complications. In many cases active CMV infection was detected in children during remission without clinical signs of uveitis activity. Individual analysis of clinical laboratory data is needed for each patient in order to evaluate the etiological and pathogenetic role of active CMV infection.

[Diagnosis of the active form of cytomegalovirus infection based on determination of IgG antibodies to the immediate-early cytomegalovirus protein by lanthanide immunofluorescence analysis]. / Diagnostika aktivnoi formy tsitomegalovirusnoi infektsii na osnove opredeleniia IgG-antitel k predrannemu belku tsitomegalovirusa metodom lantanidnogo immunofliuorestsentnogo analiza.

A system for time-resolved fluoroimmunoassay (TR-FIA) is developed for detecting early IgG during the active stage of cytomegalovirus infection on the basis of CMV-control enzyme immunoassay system. The antigen is cloned and expressed in E. coli recombinant main immediate early protein of human CMV. The active form of CMV infection was detected additionally in 5 out of 25 women who gave birth to sick babies by means of the new test system in complex with other laboratory diagnostic methods. The test is recommended for prenatal diagnosis of active CMV infection in pregnant women and for predicting the risk of neonatal disease.

[Effect of L-lysine-a-oxidase on reproduction of herpes simplex type I virus in vitro]. / Vliianie L-lisin-a-oksidazy na reproduktsiiu virusa gerpesa prostogo pervogo tipa in vitro.

L-lysin-a-oxidase (LO), a fungal enzyme catalysing oxidative deamination of L-lysin, was used for the inhibition of the reproduction of herpes simplex virus type 1 (HSV-1). Antiviral activity of LO was tested in vitro. The expression of viral antigens and CPE of HSV-1 was inhibited by LO at a concentration 0.7 mg/ml.

[Detection of herpes simplex virus by DNA-DNA hybridization method]. / Vyiavlenie virusa gerpesa prostogo metodom DNK-DNK-gibridizatsii.

Eight recombinant clones were obtained by insertion of BamHI fragments of herpes simplex type I viral DNA into a vector plasmid pUC19o. Of the obtained clones 5 were found to hybridize with herpes simplex type I and 2 viral DNA while 3 clones revealed a positive reaction with the Vero cells DNA. A constructed DNA-probe possessing the highest level of activity was selected for further studies. The probe is a BamHI fragment of herpes simplex type I viral DNA labelled with 32P dTTP. Probe sensitivity in blot hybridization is 10 pg for identification of type I viral DNA and 50 pg for type 2 viral DNA. The DNAs of cytomegalovirus and herpes zoster virus do not show positive signals with the probe. The increased sensitivity of the used dot hybridization as compared with biological or IEA antigen identification of the virus was confirmed with the clinical material from 59 patients with the different clinical manifestations of the herpes viral infection.

[The use of the indirect hemagglutination reaction for the serodiagnosis of HIV infection]. / Primenenie reaktsii nepriamoi gemaggliutinatsii dlia serodiagnostiki VICh-infektsii.

An assay system, based on the passive hemagglutination test and permitting the serodiagnosis of HIV infection with correct results in more than 99% of cases, has been developed. Three kinds of freeze-dried erythrocyte diagnostica (with shelf life exceeding 6 months), possessing high serological activity and sensitized with recombinant gene-engineering polypeptides, have been obtained. The proposed assay system is highly promising for mass examination of sera for the presence of antibodies to HIV due to the simplicity of assay techniques, the possibility of storing the diagnostica within a wide range of temperatures (4 degrees-30 degrees C) and obtaining results in a short time (3 hours).

[Structural analysis of alphoid DNA of primates. I. Heterogeneity of nucleotide sequence of alphoid repeats in human DNA]. / Strukturnyi analiz al'foidnoi DNK primatov. I. Geterogennost' nukleotidnykh posledovatel'nostei al'foidnykh povtorov DNK cheloveka.

The nucleotide sequence of two cloned fragments of human alphoid DNA was established. These fragments were earlier characterized in our laboratory as molecular markers of the 3rd (pHS05) and 11th (pHS53) chromosomes. Fragment pHS53 (2546 bp) contains alphoid repeats tandemly arranged and organized into three highly homologous pentamers. The heterogeneity of monomeric sequences within individual pentamers reaches 24-33%. Structural analysis of EcoRI subfragment pHS05 showed that this alphoid tetramer consists of two dimers 340 bp long. These dimers differ up to 16% from each other and from the so-called consensus sequence of the EcoRI-340 bp-restriction fragments family reported earlier by Wu and Manuelidis. The primary structure of four cloned fragments of EcoRI-340 bp-family was established. The data show that human alphoid DNA is highly heterogeneous. This conclusion is opposite to the view suggesting that alphoid DNA is a highly homogeneous class of reiterated sequences of the human genome.

[Structural analysis of alphoid DNA of primates. II. Evolution and possible origin of alphoid DNA of primates]. / Strukturnyi analiz al'foidnoi DNK primatov. II. Evoliutsiia i vozmozhnoe proiskhozhdenie al'foidnoi DNK primatov.

A computer analysis of human and primate alphoid DNA was performed. The number and localization of short inverted complete repeats within alphoid DNA dimers (but not monomers) remain conserved. Thus, in spite of high heterogeneity of the primary structure the conserved secondary structure of alphoid DNA might be functionally important. The analysis of internal periodicity of the monomeric sequences of human and primate alphoid DNA revealed its potential ancient sequence, that is a simple satellite DNA with a reiterated heptanucleotide TGAAAAA, which is suggested to be the ancestor of satellite DNase of rodents. The facts reported propose the ancient origin and possible functional role of alphoid-like DNA as a universal pericentromeric superfamily of DNA.

[Structural analysis of the Alu-containing DNA fragment with disperse distribution in human chromosomes]. / Strukturnyi analiz Alu-soderzhashchego fragmenta DNK s dispersnym raspredeleniem v khromosomakh cheloveka.

Nucleotide sequencing of two terminal subfragments of a cloned human DNA fragment has been done. The fragment cloned hybridized dispersively along all chromosomes except for Y chromosome and C heterochromatic chromosome regions. Both subfragments sequenced contain Alu repeats, whose structures differ partially from those of accurately determined sequences of human Alu repeats. The results obtained are discussed in respect of possible usage of Alu repeats containing sequences for construction of special polymorphic molecular markers of human chromosomes.

[Vacuum transfer of DNA to filters for detecting interindividual polymorphism by the Southern blotting hybridization method]. / Vakuumnyi perenos DNK na fil'try dlia vyiavleniia mezhindividual'nogo polimorfizma metodom "blotting"-gibridizatsii Sauzerna.

The data are provided on the possibility of detecting interindividual polymorphism from the organization of repeated DNA sequences of the alphoid type in human genome. The data were obtained on the basis of the Sothern blotting hybridization method whose efficacy was raised by means of vacuum transfer of DNA from gel to nitrocellulose filters.

[Interaction of DNA with RNA-polymerase from Escherichia coli cells infected and uninfected with T2 phage]. / Vzaimodeistvie s DNK RNK-polimerazy nezarazhennykh i zarazhennykh fagom T2 kletok Escherichia coli.

RNA polymerase from T2 infected E. coli has greatly reduced activity as compared to the enzyme from uninfected bacteria. Nevertheless both RNA polymerases synthesize heterogenous RNA species with the same maximum corresponding to chain length of 5600 nucleotides on T2 DNA. Rifampicin-challenge experiments suggest that these enzymes have identical kinetics of RNA chain initiation and elongation but their ability to form rapidly starting (RS) and delay starting (I) binary complexes with phage DNA are different. The temperature of I leads to RS transition on T2 DNA is 15 degrees for E. coli holoenzyme, but is 35 degrees for the RNA polymerase from infected cells. The transition temperature depends both on the core and on sigma fraction. Shift temperature technique was developed to investigate the kinetics of I in equilibrium RS complexes rearrangements and their temperature dependence. The rate of these rearrangements is strongly temperature dependent for E. coli holoenzyme, while for RNA polymerase from infected cells it is much lower and is practically temperature independent. From the kinetic data and from the temperature dependence of equilibrium RS-complexes concentration, the rate constants of RS-complexes formation and decay are calculated. The kinetic data obtained in rifampicin challenge experiments are in agreement with the data on the dissociation of DNA-enzyme complexes performed by the filter assay.