The aim of this study was to examine in vitro the antioxidant and antiviral activity of echinochrome A and echinochrome-based antioxidant composition against tick-borne encephalitis virus (TBEV) and herpes simplex virus type 1 (HSV-1). Materials and methods. TBEV (Dal’negorsk strain, Far Eastern subtype) grown in PK cells, and HSV-1 (VR3 strain) in Vero cells. The antioxidant activity of the compounds was determined using the linetol peroxide oxidation model. The cytotoxicity and antiviral activity of the compounds were assessed by cell viability (PK- and Vero cells) and by cytopathic effect inhibition of viruses (TBEV and HSV-1) using the MTT test. Results. The antioxidant composition, which is a mixture of echinochrome A, ascorbic acid and α-tocopherol (5: 5: 1), showed a higher antioxidant and antiviral efficacy than echinochrome A. The antiviral mechanisms on of echinochrome A and antioxidant composition are caused by direct inactivation of TBEV and HSV-1 viruses and inhibition of virus penetration into cells. Conclusion. The results obtained allow considering the echinochrome A and the composition of antioxidants on its basis as the promising agents of a broad-spectrum antiviral activity.

The aim of the work was to study the activity of echinochrome A, a naphthoquinoid pigment from sea urchins, and its antioxidant composition against herpes simplex virus type 2 (HSV-2) in vitro and in vivo.Materials and methods. Strain HSV-2 (G ATCC VR-734) was grown in Vero cells. The cytotoxic and anti-HSV-2 activity of the compounds was assessed in vitro by the cell viability and by cytopathic effect inhibition of virus using MTT test. The efficacy of compounds in mice model of vaginitis caused by HSV-2 was determined by the average lifetime, body weight and viral load changes.Results and discussion. The antioxidant composition (echinochrome A, ascorbic acid and α-tocopherol (5:5:1)), showed a higher antiviral efficacy than echinochrome A alone. Oral administration of the antioxidant composition protected 90% of the infected mice against death and reduced vaginal viral loads. The antiviral activity of echinochrome A and the antioxidant composition is probably due to the virus-inhibiting activity of the compounds and their antioxidant properties.Conclusion. The results obtained allow considering the tested compounds as promising agents with antiviral properties.

The aim of this study was to examine in vitro the antioxidant and antiviral activity of echinochrome A and echinochrome-based antioxidant composition against tick-borne encephalitis virus (TBEV) and herpes simplex virus type 1 (HSV-1). Materials and methods . TBEV (Dal’negorsk strain, Far Eastern subtype) grown in PK cells, and HSV-1 (VR3 strain) in Vero cells. The antioxidant activity of the compounds was determined using the linetol peroxide oxidation model. The cytotoxicity and antiviral activity of the compounds were assessed by cell viability (PK- and Vero cells) and by cytopathic effect inhibition of viruses (TBEV and HSV-1) using the MTT test. Results . The antioxidant composition, which is a mixture of echinochrome A, ascorbic acid and α-tocopherol (5: 5: 1), showed a higher antioxidant and antiviral efficacy than echinochrome A. The antiviral mechanisms on of echinochrome A and antioxidant composition are caused by direct inactivation of TBEV and HSV-1 viruses and inhibition of virus penetration into cells. Conclusion . The results obtained allow considering the echinochrome A and the composition of antioxidants on its basis as the promising agents of a broad-spectrum antiviral activity.

AIM:Comparative evaluation of effectiveness of traditional serologic and modified diagnostic methods of disease arising due to varicella and herpes zoster virus (VZV) reactivation.MATERIALS AND METHODS:2 groups of patients were examined. The main group consisted of 39 patients with manifest form of herpes zoster (HZ), control--20 healthy donors. Sex composition of the groups did not differ. Traditional method of serologic diagnostics included determination of anti-gE VZV IgG and anti-VZV IgG and anti-IgM in patient and donor blood sera by using EIA. Modified methods consisted of isolation in density gradient and cultivation for 48 hours of peripheral blood mononuclears (PBMC) in RPMI-1640 complete culture medium containing 10% of fetal bovine serum, 4 mM L-glutamin and gentamycin. Concentrations ofanti-VZV IgG and IgM were then determined in culture medium by using EIA.RESULTS:In all the examined HZ patients and healthy donors anti-VZV IgG were detected in blood. Only in 26 (67%) of 39 HZ patients anti-gE VZV IgG and anti-VZV IgM were determined in blood sera. Among donors false positive results for these markers were detected in 10% and 5% of cases, respectively. During simultaneous determination of anti-gE VZV IgG and anti-VZV IgM the specificity of the method increased to 100%, sensitivity of the diagnostic method based on simultaneous determination of anti-gE VZV IgG and anti-VZV IgM was 59%. During analysis of spontaneous production of anti-VZV antibodies by PBMC in 38 (97.4%) of 39 patients anti-VZV IgG were determined in PBMC culture, anti-VZV IgM production was observed only in 4 patients. In control group false positive results of anti-VZV IgG and IgM production by PBMC was not detected by the modified method (100% specificity). At equal specificity level sensitivity of the modified method based on determination of spontaneous anti-VZV IgG production by PBMC culture was significantly higher than effectiveness of the traditional serologic diagnostics (97.4% and 59%, p < 0.0001).CONCLUSION:The data obtained allow to recommend during diagnostics of manifest and atypical VZV infection forms arising due to endogenous virus reactivation the new modified method of laboratory diagnostics of the disease as having higher sensitivity compared with traditional serologic method.

The new Russian enzyme immunoassay system "CMV-Diagnost" based on the detection of low-avid IgG antibodies has been developed for the rapid diagnosis of cytomegalovirus infection. The system was found not only to determine the strained immunity in response to cytomegalovirus, but also to judge the current infection from the avidity index of detectable IgG antibodies with a high degree of validity. The antibody avidity index of less than 30% suggests an acute stage of primary cytomegalovirus infection. The minimum antibody threshold bodies (deltaOD has been established for the correct interpretation of data on low-avid antibodies. deltaOD of > or =0.6 optic units was for the developed test system "CMV-Diagnost. A correlation was found between the serum levels of low-avid antibodies and IgM antibodies to cytomegalovirus at the acute stage of the disease.

The antiviral action of a natural cytokine complex (NCC)--the preparation Superlymph and its peptide antimicrobial fraction (AMF)--in the culture of Vero cells infected with type 1 herpes simplex virus (HSV-1), strain VR-3, was studied. The NCC preparation did not alter the morphology of the cells for 6 days and was not toxic for the culture of Vero cells. The NCC and AMF produced a protective antiviral effect, which was manifested by the inhibition of the cytopathic action (CPA) of the virus. In the presence of the preparation, the CPA of HSV-1 was equal to 10(-4.67) ICPD50, while in the control CPA was equal to 10(-5.60). The fraction containing antimicrobial peptides (protegrins) and isolated from NCC, characterized by the method of mass spectrometry, produced the maximum antiviral effect on the cell strain Vero (10(-4.58) ICPD50). Thus Superlymph, an immunomodulator with antiviral activity, could be regarded as an effective preparation for the treatment of HSV infection. The action of such preparation was aimed at the inhibition of the CPA of the virus and the stimulation of the antiviral protective mechanisms of the cell.

A Russian immune-enzyme test-system ("HERPES-DIAGNOST") was designed on the basis of detection of low-avidity IgG antibodies in order to promote the laboratory value of serological examinations of patients with different clinical manifestations of the herpetic infection. The key test parameters were tuned; the immunosorbent production based on antigens (herpes simplex virus--HSV), types 1 and 2, was optimized; and the concentration was chosen for the main reagent that removes the low-avidity antibodies (8 M urea solution) and, finally, the temporal and temperature regimes were selected for testing. A system was elaborated for registering and interpreting the results. The avidity index of antibodies lower than 35% was found to be a reliable criterion confirming the presence of acute primary infection triggered both by HSV-1 and by HSV-2. If there is a relapse of herpetic infection, the avidity index of antibodies can range from 30 to 45%.

Examinations of 202 newborn babies for a representative group of viral infections by detection of viral antigens in cells of urine sediment and in the autopsy materials by indirect immunofluorescence permitted diagnosis of a congenital viral infection in 92% of patients with intrauterine and perinatal pathology; in 72.5% it was a mixed infection. In the patients the virus-virus associations were, as a rule, represented by enteroviruses of Coxsackie group and/or influenza A, B, and C viruses. Most frequently (83.3-100%) mixed virus infection was detected in newborn babies with the severest pathology (meningoencephalitis, encephalitis, sepsis, intrauterine pneumonia), as well as in fatal cases.

Examinations of 202 newborn babies for a representative group of viral infections by detection of viral antigens in cells of urine sediment and in the autopsy materials by indirect immunofluorescence permitted diagnosis of a congenital viral infection in 92 % of patients with intrauterine and perinatal pathology; in 72.5 % it was a mixed infection. In the patients the virus-virus associations were, as a rule, represented by enteroviruses of Coxsackie group and/or influenza A, B, and C viruses. Most frequently (83.3-100 %) mixed virus infection was detected in newborn babies with the severest pathology (meningoencephalitis, encephalitis, sepsis, intrauterine pneumonia), as well as in fatal cases.

The authors analyze the relationships between various forms of lupus erythematosus with a chronic viral infection. Three cases with cicatrizant erythema associated with high titers of antiviral antibodies are discribed.

The authors demonstrate a high sensitivity and specificity of an enzyme immunoassay test system for the diagnosis of genital herpes.

The results are presented of comprehensive virological studies and clinical description of a 10-year-old boy suffering from atopic dermatitis from the age of 1 month. Oral and external alpizarin therapy of herpes was fairly effective.

The data on the use of a commercial EIA test system for detection of antibodies in control of preparations against herpes simplex and cytomegaloviruses are presented. The enzyme immunoassay test system for antibody determinations to herpes simplex virus produced by the Odessa bacterial preparations enterprise was shown to be suitable for determination of the specific potency (antigenicity) of herpes simplex vaccine. The advantages of this method over the currently used neutralization test were established. Titration of commercial immunoglobulins detects lots with high litres of antibody to herpes simplex virus. For the same purpose, lots of commercial immunoglobulins were tested for antibodies to cytomegalovirus using a West Germany test-system (Behring). It is concluded that enzyme immunoassay test systems for antibody determinations may be used for screening of lots of immunoglobulins of special effects (against herpes simplex and cytomegalovirus infections) both at the stage of serum and final preparation screening.

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 data on the use of a commercial EIA test system for detection of antibodies in control of preparations against herpes simplex and cytomegaloviruses are presented. The enzyme immunoassay test system for antibody determinations to herpes simplex virus produced by the Odessa bacterial preparations enterprise was shown to be suitable for determination of the specific potency (antigenicity) of herpes simplex vaccine. The advantages of this method over the currently used neutralization test were established. Titration of commercial immunoglobulins detects lots with high litres of antibody to herpes simplex virus. For the same purpose, lots of commercial immunoglobulins were tested for antibodies to cytomegalovirus using a West Germany test-system (Behring). It is concluded that enzyme immunoassay test systems for antibody determinations may be used for screening of lots of immunoglobulins of special effects (against herpes simplex and cytomegalovirus infections) both at the stage of serum and final preparation screening.

: The result obtained in the study of the possibility of using the method for the determination of the titer of antibodies to herpes simplex virus by EIA techniques in a single dilution of the serum under test are presented. This method is based on the determination of the optical density of the serum titer (rcut) in different groups of sera with the use of the assay system, permitting the evaluation of the positive results obtained in the determination of their final dilution. The results obtained with the use of this method showed that error was 50% for high-titer sera, 60% for medium-titer sera and 30% for low-titer sera.