[Antiviral activity of the organic germanium complex with aciclovir against herpes simplex virus (Herpesviridae: Alphaherpesvirinae: Simplexvirus: Human alphaherpesvirus 1/2) in the in vitro and in vivo systems].

INTRODUCTION: A significant increase in the incidence of various forms of herpesvirus infection (HVI) determines the need to search for new approaches to the modification of one of the basic antiviral drugs aciclovir (ACV) and its dosage forms to improve their biopharmaceutical characteristics and increase the effectiveness of therapy. In this aspect, an innovative organic germanium complex with aciclovir (OGCA) is promising.The aim of the study was to assess the antiviral activity of OGCA against the herpes simplex virus (HSV) (human herpes virus, HHV) on the HVI models both in vitro and in vivo. MATERIAL AND METHODS: We studied the activity of OGCA in a therapeutic regimen against HSV-1 (HHV-1) (Kl strain), HSV-2 (HHV-2) (VN strain) using virological and statistical research methods in the in vitro model of HVI on Vero cell culture and the model of genital herpes (GH) caused by HHV-2 (VN strain) in male guinea pigs (Canis porcellus). RESULTS AND DISCUSSION: It was found OGCA inhibits the replication of HHV-1 and HHV-2 in Vero cells, and has anti-HHV activity in the GH model in male guinea pigs, leading to a decrease in the severity and duration of the disease, the intensity and duration of viral shedding. The most pronounced activity was detected when preparation was applied topically 5 times a day for 5 days at the early stages of infection (3% gel). The delayed use of OGCA (48 hours after infection) also had statistically significant efficacy compared to commercial reference drugs containing aciclovir or its pro-drugs: aciclovir (5% cream), AIL (acyclovir+interferon alfa-2b+lidocaine, 3% ointment), penciclovir (1% cream). OGCA significantly reduced the number of days of the pathogen shedding, as well as its infectivity, compared to animals in the control group and ones receiving placebo. The activity of OGCA, apparently, is due to its improved biopharmaceutical characteristics compared to aciclovir, as well as the presence of a number of biological activities of its constituent components. CONCLUSION: The results of the study allow us to consider OGCA as the basis for the development of antiviral agents for the treatment of HVI.

[Study of the antiviral activity of the liquid corneal storage medium in relation to the herpes simplex virus in vitro].

The aim of the study was to assess the antiviral activity of the 7 types of liquid corneal storage medium on an experimental model of herpesvirus infection in vitro. MATERIAL AND METHODS: The study of antiviral activity of 7 samples of liquid corneal storage medium on a model of herpesvirus infection caused by the herpes simplex virus type 1 in Vero-cell using virological and statistical research methods was carried out. RESULTS AND DISCUSSION: All the studied images of the corneal storage medium, including the Borzenka-Moroz base medium, did not have a cytotoxic effect on Vero cell culture. Out of of 7 samples of liquid corneal storage medium, 4 samples had reliable antiviral activity against HSV-1 when used under the therapeutic regimen (1 hour after infection) and under the preventive regimen (2 hours before infection). Antiviral activity was established in 2 samples containing the interferon inducer cycloferon at a concentration of 10 mg/kg and 30 mg/kg (sample 2, 3), in a sample containing the interferon inducer gamapren 15 mg/kg (sample 5), and in a sample containing a combination of drugs - 10 mg/kg cycloferon and an acyclic nucleoside analog-acyclovir 10 mg/kg (sample 6). According to the results of 2 test regimens, the maximum statistically significant inhibitory effect in relation to HSV-1 was detected in sample 6, containing a combination of drugs. Against the background of sample 6, the infectious activity of the test virus decreased by an average of 3.2 lg, the inhibition coefficient was 54.5%. CONCLUSION: The results of the study indicate the prospects of using types of media with antiviral activity (samples 2, 3, 5, 6) for storing donor corneas in order to increase the effectiveness of keratoplasty in patients with ophthalmic herpes.

[THE SPECTRUM OF MARKERS OF HERPES VIRAL INFECTIONS AND ALGORITHM OF THEIR LABORATORY DIAGNOSTIC IN CHILDREN WITH INFLAMMATORY PROCESSES OF UPPER RESPIRATORY WAYS AND ENT-ORGANS].

The purpose of study is to explore markers of persistent herpes viral infections in children with inflammatory processes of upper respiratory ways and ENT-organs. The sampling included 118 examined patients aged from 1 month to 17 years. The complex of standardized viral, immunological, molecular genetic methods was applied to detect (to exclude) herpes infection: cytomegalovirus infection, Epstein-Barre virus infection, simplex herpes virus infection. The diagnostic algorithm of examination of children with diseases of upper respiratory ways for herpes infection is presented. The dominating significance of simplex herpes virus and Epstein-Barre virus and also Staphylococcus aureus and Streptococcus haemolyticus-ß group A at the analysis of microbial landscape. In 83.9% of children with diseases of upper respiratory ways chronic infections of simplex herpes virus, Epstein-Barre virus, cytomegalovirus; in39.39% - mixed-infection; in 41.03% - combination of simplex herpes virus and Epstein-Barre virus infections; in 33.33% - combination of simplex herpes virus and cytomegalovirus infections; in 7.69% - combination of simplex herpes virus and Epstein-Barre virus and cytomegalovirus infections; in 17.94% - combination of Epstein-Barre virus and cytomegalovirus infections; The particularity of course of persistent herpes infection in children had to do with absence of specific symptoms of nosologic form in 59.2% of cases. The results of analysis of smears from nasopharynx of children infected with herpes viruses permitted to detect: Staphylococcus aureus in 36.36%; Streptococcus haemolyticus-ß in 32.32%; Streptococcus haemolyticus-α in 11.11%; Candida albicans of mucous membranes in 4.04% of children. The viral bacterial mixed-infection was detected in 44.44%. The laboratory signs of activity of immune inflammation were detected: increasing of content of TNАα and decreasing of level of IFNγ. The results of study substantiate necessity of individual approach to therapy of children with diseases of upper respiratory ways and ENT-organs and with implementation of complex of curative rehabilitating activities.The purpose of study is to explore markers of persistent herpes viral infections in children with inflammatory processes of upper respiratory ways and ENT-organs. The sampling included 118 examined patients aged from 1 month to 17 years. The complex of standardized viral, immunological, molecular genetic methods was applied to detect (to exclude) herpes infection: cytomegalovirus infection, Epstein-Barre virus infection, simplex herpes virus infection. The diagnostic algorithm of examination of children with diseases of upper respiratory ways for herpes infection is presented. The dominating significance of simplex herpes virus and Epstein-Barre virus and also Staphylococcus aureus and Streptococcus haemolyticus-ß group A at the analysis of microbial landscape. In 83.9% of children with diseases of upper respiratory ways chronic infections of simplex herpes virus, Epstein-Barre virus, cytomegalovirus; in39.39% - mixed-infection; in 41.03% - combination of simplex herpes virus and Epstein-Barre virus infections; in 33.33% - combination of simplex herpes virus and cytomegalovirus infections; in 7.69% - combination of simplex herpes virus and Epstein-Barre virus and cytomegalovirus infections; in 17.94% - combination of Epstein-Barre virus and cytomegalovirus infections; The particularity of course of persistent herpes infection in children had to do with absence of specific symptoms of nosologic form in 59.2% of cases. The results of analysis of smears from nasopharynx of children infected with herpes viruses permitted to detect: Staphylococcus aureus in 36.36%; Streptococcus haemolyticus-ß in 32.32%; Streptococcus haemolyticus-α in 11.11%; Candida albicans of mucous membranes in 4.04% of children. The viral bacterial mixed-infection was detected in 44.44%. The laboratory signs of activity of immune inflammation were detected: increasing of content of TNАα and decreasing of level of IFNγ. The results of study substantiate necessity of individual approach to therapy of children with diseases of upper respiratory ways and ENT-organs and with implementation of complex of curative rehabilitating activities.

[Evaluating the effectiveness of Solanum tuberosum shoots extract in experimental ocular herpes].

UNLABELLED: Ocular herpes (OH) is an infectious disease caused by the herpes simplex virus (HSV) characterized by a variable clinical presentation and often accompanied by complications that may lead to deterioration of visual functions, cataract development, or even blindness. Its treatment is arduous. The aim of this work was to evaluate the effectiveness, tolerability, and safety of Panavir eye drops in a rabbit model of OH. MATERIAL AND METHODS: Ocular infection was induced with HSV-1 (EU strain) in grey rabbits (all males, 2.5-3.0 kg) according to the standard technique. The treatment included Panavir-GLA (Panavir-gamma-linolenic acid) and Panavir medications. RESULTS: Panavir eye drops instilled 6 times daily for 8 days showed a pronounced therapeutic effect and prevented the development of severe corneal opacities. The most rapid and significant results were seen in rabbits with epithelial keratitis and those with short-term persistence of the virus. Generally, the effectiveness of Panavir eye drops was comparable with that of the reference drug (Oftalmoferon). Panavir instillations caused no irritation, toxic and/or allergic effects and were well tolerated by the rabbits. CONCLUSION: The data obtained suggest that Panavir eye drops may be included in OH treatment schemes.

[Results of using Solanum tuberosum sprouts extract for the treatment of ophthalmoherpes].

The efficacy, tolerability and safety of the extract of Solanum tuberosum sprouts (Panavir eyedrops) have been studied on the model of ophthalmic disorder in rabbits caused by herpes simplex virus (HSV) type 1. It is established that Panavir applied via 6 instillations per day for a period of days has potent therapeutic efficacy and prevents the development of gross corneal opacity in rabbits. Instillation of Panavir eyedrops does not cause irritation, toxic and allergic effects and are well tolerated by rabbits. The fastest and most pronounced effect of Panavir eyedrops was observed in the treatment of epithelial keratitis, as well as for not prolonged persistence of HSV. The effectiveness of Panavir eyedrops is comparable with that of the reference preparation OphthalmoferonR.

[Cycloferon and management of herpes virus infection].

The treatment of patients with various forms of herpes requires a complex approach with using chemo- and immunotropic drugs. The use of Cycloferon, an interferon inductor (12.5% injection solution, 150 mg tablets or 5% liniment) was shown efficient. It had antiviral and immunotropic action in the mono- and combination therapy of herpes simplex of the skin and mucosa, genital herpes, ophthalmoherpes, herpes zoster, infectious mononucleosis. Cycloferon lowered the level and period of the disease clinical signs, prolonged the remission, corrected the immunity shifts, prevented the complications. The results of the study presented a conclusive proof for recommending such a use of Cycloferon in wide medical practice.

[Vaccines as an approach to the immunocorrection in herpetic infections].

Development of vaccines for immunologic correction in herpetic infections is an important problem that raises a growing concern worldwide. The data on the experimental studies of the efficacy of an inactivated whole-virion vaccine against herpes simplex viruses types 1 and 2 (HSV-1 and -2) using an animal model are discussed. The results of the multiyear application of the vaccine to ophthalmology and dermatology practice are also presented. The results unambiguously show a high efficacy of the vaccine in the prevention of recurrences of the infections based on activation of specific T-cell response. A live vaccine against the varicella zoster virus (VZV) was developed for control of the infection in children. For the cytomegalovirus (CMV) infection in adults, inactivated whole-virion vaccines are at the stage of development. An important part of the study addresses a combined application of the inactivated vaccines with immunomodulators.

[Immunomodulators and specific inactivated vaccines in urgent prophylaxis under experimental arboviral infection].

A reliable protective activity of the home-manufactured immunomodulators (ridostin, polyribonate glucosemuramyl-dipeptide, Mylife, and peptidoglycane-160) was detected in mice. The mice were infected with the equine eastern encephalomyelitis virus (EEEV, an alphavirus), or with the tick-borne encephalitis virus (TBEV), or the yellow fever (YF) virus (both flaviviruses). The effect of the urgent vaccination reliably increases when the vaccination is combined with the immunomodulators listed above. Under the alphavirus infection, the combined effects of the vaccine and ridostin were accompanied with increased specific humoral and cellular immune response (virus-specific antibodies and adoptive transfer of immune lymphocytes). The combined application of the specific vaccine and ridostin can be recommended for clinical trials of TBE in the foci of Infection.

[Efficiency of coadministration of immunomodulators and vaccine in an experiment on tick-borne encephalitis].

Experiments on a tick-borne encephalitis (TBE) model in CBA and BALB/c mice demonstrated that immunomodulators (ridostin, polyribonate, and peptidoglycan-160) and a specific vaccine against TBE were significantly effective in increasing the level of a protective effect and life expectancy in the experimental group as compared to the control group. The findings allow one to recommend the immunomodulator ridostin in combination with the inactivated vaccine for the emergency prophylaxis of TBE in its virus-infected subjects in the foci of infection.

[Combined effects of immunomodulators and specific inactivated vaccines against alpha- and flavivirus infections].

The combined effects of immunomodulators, such as ridostin, polyribonate, glucose muramyl dipeptide, and peptidoglycan-160, and specific vaccines on survival of mice with alpha- (eastern equine encephalitis) and flavivirus (tick-borne encephalitis (TBE) infections were found to be significant. In alphavirus infection, the combined effects of the specific vaccine and ridostin were accompanied by increases in specific humoral immunity (specific antibodies) and cellular immunity (adoptive transfer of immune lymphocytes). The concurrent use of the specific vaccine and the immunomodulator ridostin is recommended in clinical trials of TBE in the foci of infection.

Functional activity of peritoneal macrophages from sensitive and resistant mouse strains during intravaginal infection with herpes simplex virus type 2 and mucosal vaccination.

In the early period after intravaginal infection with herpes simplex virus type 2 (2 h), macrophages from sensitive DBA/2 mice were characterized by higher capacity to engulf the antigen, decreased function of the lysosomal apparatus, lower activity of cathepsin D, and reduced oxygen metabolism compared to cells from resistant BALB/c mice. Mucosal vaccination with herpes vaccine and hyaluronic acid promoted the increase in functional activity of macrophages and improved survival of sensitive mice (by 60%).

Immunostimulating activities of the novel peptidomimetic L-glutamyl-histamine.

An original representative of histamine-containing peptidomimetics L-glutamyl-histamine (L-Glu-Hist) was synthesized and characterized as a cytokine mimic leading to cellular responses of improved specificity. The energy-minimized 3-D conformations of L-Glu-Hist derived from its chemical structure resulted in stabilization for Fe(2+) chelating complexes. L-Glu-Hist accelerated the decrease of ferrous iron in the ferrous sulphate solution in a concentration-dependent mode and showed the ferroxidase-like activity at concentrations less than 3 mm in the phenanthroline assay, whereas in the concentration range 3-20 mm L-Glu-Hist restricted the availability of Fe(2+) to phenanthroline due to binding of ferrous ions in chelating complexes. L-Glu-Hist showed a stimulatory effect on phosphatidylcholine liposomal peroxidation (LPO) catalysed by the superoxide anion radical (O(2) (*))-generating system (Fe(2+)+ ascorbate) at low (less or about 1 mm) L-Glu-Hist concentrations and both revealed the inhibitory effect on LPO in this system of high ( approximately 10 mm) L-Glu-Hist concentration. L-Glu-Hist released O(2) (*) in concentrations which stimulated [(3)H]-thymidine incorporation into DNA and proliferation of mouse spleen lymphocytes and mononuclear cells from human blood. The structural peptide-like analogues of L-Glu-Hist such as L-Glu-Trp, carcinine (beta-alanylhistamine), but not L-Pro-Glu-Trp were active in stimulating thymidine incorporation and in inducing proliferation of mononuclear cells compared to mitogen concanavalin A at doses 2.5-25.0 microg/ml. Our data provide evidence that L-Glu-Hist may act as a very fast and sensitive trigger for lymphocyte proliferation and immunoregulation.

[Antiviral activity of human recombinant gamma-interferon and recombinant hybrid protein of tumor necrosis factor-alpha-thymosin-alpha1 on models of herpes-virus and cytomegalovirus infection in vitro]. / Protivovirusnaia aktivnost' rekombinantnogo interferona-gamma cheloveka i rekombinantnogo gibridnogo belka alpha-faktor nekroza opukholei-timozin-alpha1 na modeliakh gerpes-virusnoi i tsitomegalovirusnoi infektsii in vitro.

The conducted studies showed a certain efficiency of gene-engineering preparation of IFN-gamma and TNF-T in virus infections caused by herpes simplex virus 2 (HSV-2) and cytomegalovirus (CMV) in cell cultures of human embryo fibroblast (HEF). The drugs have no viricidal action. IFN-gamma, when used according to the treatment scheme in vitro, proved to be more effective versus TMF-T both in HSV-2 and in CMV. It inhibited significantly the HSV-2 reproduction within the dilution range of 1:50 to 1:500. It was also effective, when used in CMV, within the dilution range of 1:5000 and lower, whereas TNF-T was effective within the range of 1:500 and lower as well as in 0.1 multiple infection. A significantly higher effect was ensured when the drugs were used for prevention. In HSV-2, IFN-gamma inhibited the virus reproduction, like in the treatment scheme, within the dilution range of 1:50 to 1:500, whereas TNF-T was effective in the range of 1:50. In CMV, the drugs' effect, when used for prevention, was similar to that observed in the treatment scheme. The highest inhibition values were registered for HSV-2, when it was used 24 hours before infection (IFN-gamma--2.25 Ig, dilution range of 1:50; TNF-2--1.0 Ig, dilution range of 1:50). IFN-gamma and TNF-2 exert a synergic action on different stages of virus reproduction. A reliable additive effect was ensured in prevention made 4 hour before infection by IFN-gamma and TNF-T only in experimental CMV infection.

[Polymorphism of herpes simplex virus type 1 and 2 DNA from laboratory strains and clinical material from patient with genital herpes]. / Polimorfizm DNK virusa prostogo gerpesa tipov 1 i 2 iz laboratornykh shtammov i klinicheskogo materiala ot patsientov s genital'nym gerpesom.

Polymerase chain reaction (PCR), that can amplify a fragment of the DNA-polymerase gene of 4 herpes viruses, i.e. herpes simplex viruses, type 1 (HSV-1), herpes simplex viruses, type 2 (HSV-2), Epstein-Barr virus and cytomegalovirus, was made use of to study the genetic polymorphism of HSV-1 and HSV-2 strains. The obtained amplicons were analyzed by the method of restriction-size fragments' polymorphism (RSFP) with restrictases Rsal, Taql and Hinfl. Four HSV-1 strains had an identical restriction profile. Strain G (HSV-2) also displayed the expected restriction profiles, however, contradictory results were obtained for strain BH (HSV-2): the restriction profiles with restrictases Hinfl and Rsal corresponded to HSV-2, and the restriction profile with Taql corresponded to HSV-1. The sequencing of appropriate fragments of strains G and BH revealed a dot-type mutation localized in Taql restriction site. The thus worked out PCR was used jointly with RSFP in the genotyping of 75 urogenital samples obtained from women with genital herpes who were treated at Moscow patient-care facilities. HSV-1 and HSV-2 were detected in 18 (24%) and 57 (76%) of samples, respectively. No changes were registered in the restriction profile for HSV-2 among the investigated samples and all of them had the restriction profile similar to that of strain G. The conclusion is that genital herpes associated with HSV-2 is genetically stable within its Moscow population.

[The antiviral activity of peptide immunomodulator "Gepon" in experimental infections caused by herpes simplex viruses types 1 and 2]. / Protivovirusnaia aktivnost' peptidnogo immunomoduliatora "Gepon" pri éksperimental'nykh infektsiiakh, vyzvannykh virusami prostogo gerpesa tipov 1 i 2.

Experimental data are reported on the antiviral activity of peptide immune-modulator "Gepon" in infections caused by herpes simplex virus (HSV), types 1 and 2, in vitro and in vivo. The drug proved to be non-toxic and did not possess any viricidal action. Its antiviral effect was registered in experiments with a multiple-infection cells' culture. The maximal effect was a 100-fold reduction of the viral titer when it was used in a concentration of 6.25 mcg/I as a preventive measure 24 hours before triggering the infection. The mentioned drug's effect was reliably higher than its use within a treatment scheme (1 hour after the infection onset). "Gepon" possessed the reliable protecting qualities (36% of protection with a mean increase of the infected mouse life by 1.9 days) in experiments with intraperitoneally infected mice (10 LD50/mouse of HSV, type 2), when the drug was administered in doses of 0.1 and 1 mcg/mouse.

[Virus isolated in the human blood leukocyte culture and its interaction with hepatitis C and G viruses]. / Virus, vydelennyi na kul'ture leikotsitov krovi cheloveka, i ego vzaimootnoshenie s virusami gepatitov C i G.

Electrone-microscopic investigations are indicative of that the cultures of healthy donors, stimulated by phytohemagglutinin (PHA), can be successfully used to study the etiology of parenterally transmitted hepatitis. An electronic-microscopic study of a virus, isolated from the blood serum of a patient with hepatitis on the basis of the PHA-stimulated human leukocyte cultures and named a hepatitis leukocytic virus (HLV), enabled, by using the negative contrasting method, to detect viral particles of the hexagonal shape, sized 50-65 nm, with a coating divided by a 4-5 nm light space. Therefore, the HLV was described as belonging to the Flaviviridae family. RNA of the C hepatitis virus was detected in the K HLV strain stored, for 24 years, at the Museum of the Viruses Research Institute, Russian Academy of Medical Sciences, in a lyophilizated bed at -5 degrees C, however, an attempt to genotype the RNA failed. No RNA donor leukocytes were found in the materials of further passing of HLV by using the PHA-stimulated cultures, which can be explained by an inactivation of HLV at storage. No RNA of the C hepatitis virus was found in the above materials either, however, in 1999, DNA of the TT virus was detected at passing the strain, which indicates that the virus is widely spread in the population of healthy donors, whose lymphocytes are used preparing the blood leukocyte cultures.

[Therapeutic effect of gefin, a new antiviral drug, in experimental genital herpes ]. / Lechebnoe deistvie novogo protivovirusnogo preparata gefina pri éksperimental'nom genital'nom gerpese.

Antiviral activity of Gefin was studied in guinea pigs infected with herpes simplex virus (HSV-2). Up to 54.9% animals challenged with HSV survived after 7-day treatment with Gefin (3 local applications a day). The prospects of further trials of the drug in HSV genital herpes are discussed.

[Combined use of killed vaccines and immunomodulator ridostin for urgent prevention of epidemic stomatitis, Aujeszky disease and carnivore plague in experiment]. / Sochetannoe ispol'zovanie ubitykh vaktsin i immunomoduliatora ridostina dlia ékstrennoi profilaktiki iashchura, bolezni Aueski i chumy plotoiadnykh v éksperimente.

Results of experimental studies of mice and pigs infected with foot-and-mouth disease virus, minks infected with Aujeszky's disease virus, and dogs infected with canine distemper virus are described. In animals with foot-and-mouth disease and Aujeszky's disease, combined treatment with killed vaccine and immunomodulator Ridostin by the scheme of urgent prophylaxis (3 days before infection) caused 75% (foot-and-mouth disease) and 100% (Aujeszky's disease) prevention of animal death and development of generalized infection. The use of Ridostin by the scheme of urgent prophylaxis in a canine distemper infection focus arrested clinical symptoms of the disease in 50% of animals received immunomodulator. Clinical symptoms of canine distemper in the other dogs treated with immunomodulator were manifested in a mild form, and their appearance was delayed to 23-25 days after contact with infected animal.

[Development of a suppository form of the interferon inducer poludan and potentiation of its action by hyaluronic acid]. / Razrobotka svechevoi formy induktora interferona poludan i potentsirovanie ego deistviia gialuronovoi kislotoi.

A promising dosage form of interferon inductor poludan has been developed for the treatment of patients with genital herpes and ophthalmic herpes: suppositoria. High interferon-producing and antiviral activity of poludan in suppositoria is potentiated by hyaluronic acid and antioxidants.