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NEW HSV 1 TARGETS OF ACiCLOVIR ANd ITS METAL COMPLEXES

T. Varadinova, P. Genova, A. Terron, A. Garcia-Rasso, J. Fiol
Laboratory of Virology, Sofia University, Bulgaria
Department de Quimica, Universitat de les Illes Balears, Palma de Mallorca, Spain

Previously we have shown that when acetylated (Ac-ACV) or bond to Zn(II) ion the anti-HSV activity of ACV is increased and the existing to ACV resistance was overcome. Here we present data on some new virus but not cell specific targets of ACV and its derivatives. Experiments were done on HSV 1 infected MDBK cells treated with ACV, Ac-ACV, Zn-Ac-ACV, Zn(NO3)-ACV, Zn(Cl)-ACV or Ag-ACV. Virus specific proteins were identified on the 15 h by Western Blot analysis. The induction of apoptosis was determined on the 8 h by acridine orange and Janus green B dye staining. Eleven virus specific proteins were identified in viral control: VP5, VP22, VP22/22a, aTIF, VP11/12, gB, gC, gE, gD, gH and gG. In samples treated with ACV and its metal complexes VP22/22a, aTIF, VP11/12, gG/gD and gH were not identified while under the action of Ac-ACV and Zn-Ac-ACV, aTIF, VP11/12, gG/gD, gH, gE and gC were absent. These data suggest that ACV and its derivatives also suppress the morphogenesis, cell-to-cell spread and transactivation of virus genomes. These effects strongly depended on the modifications in ACV structure, as the most active against HSV 1 replication were Zn-Ac-ACV followed by Ac-ACV. All the compounds tested did not induce apoptosis both in infected and in noninfected cells. (COST D20, 0006-01)

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APPLICATION OF HERPETIC VACCINA FOR THE PREVENTION OF HERPETIC INFECTION RELAPSES

V.M. Semenov, N.F. Akulich
Vitebsk State Medical University, Republic Belarus

The aim of our study was to assess the efficiency of herpetic vaccina for the prevention of herpetic infection relapses. 200 patients ill with relapsing herpetic infection aged from 12 to 75 (136 women and 64 men) underwent our supervision. Herpes genitalis was revealed in 62% of patients, herpes labialis and nasalis – in 14%, 24% of cases were the disseminative forms of the disease. Antibodies to herpes viruses of 1st and 2nd types were detected by application of ELISA. Antibodies to type 1 herpes virus were found in 98.5% of patients, and to type 2 herpes virus – in 69%. 1.5% of patients didn’t conent antibodies to any type of herpes viruses.
Herpetic vaccina containing inactivated antigens of herpes simplex viruses of 1st and 2nd types was injected intramuscularly in a volume of 0.2 ml once per four days, totally 5 injections. The criteria of vaccinaion efficiency were the decrease of incidence and duration of relapses, the prolongation of remissions, the decrease of the rash elements quantity, disappearing of itch, burning and pain. We have achieved the absence of relapses for the time of follow-up (3-10 years) in 61% of patients. In patients having antibodies to one type of virus vaccination was effective in 89.5% of cases; in patients with antibodies to both virus types – in 50.5% only. The people with antibodies to both virus types composed 89% of persons who didn’t develop immunity. Thus, herpetic vaccina is efficient in 61% of patients. The absence of constant effect was seen in patients infected with both types of herpes simplex virus.

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STUDY OF THE NATURAL ARONIA MELANOCARPA JUICE FOR ANTI-VIRAL ACTIVITY

V. Russev, S. Valcheva-Kuzmanova, A. Belcheva
Pharmacology Section, Department of Preclinical and Clinical Pharmacology and Biochemistry, Varna Medical University, Varna, Bulgaria
Department of Microbiology and Virology, Varna Medical University, Varna, Bulgaria

The natural juice from Aronia melanocarpa fruit is rich in polyphenols most of which are flavonoids (mainly anthocyanins) and tannins. These products and other natural compounds isolated from plants are recently evaluated for their anti-infectious activity. Certain perspectives for finding new active antiviral factors are promising. The aim of the present study is to investigate the effect of Aronia melanocarpa natural juice on the reproduction in ovo of Influenza virus type A (H3N2). A concentration of 2 000 mg%/100 ml, with initial pH 3.76, is used for the experimental trial. Further in the experiments we apply pH 7.00 to pH 7.50, which corresponds to normal human pH. The results show that the Aronia juice, with concentration and pH, as indicated before, inhibits the reproduction of Influenza virus in its initial stages. The effect most probably is due to the formation of complex compounds between the virion, from one hand, and the flavonoids and tannins, from the other hand. The authors presume that this result influences the adsorption of Influenza virus on the cell surface.

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NEW AUTOMATED MONITOR FOR DIRECT VIRUS-CELL INTERACTION MONITORING

O.P. Fedchuk, A.O. Fedchuk, A.S. Fedchuk, P.O. Fedchuk
I.I. Mechnikov Odessa National University, Odessa, Ukraine
Ukrainian I.I. Mechnikov Anti-Plague Research Institute, Odessa, Ukraine
Odessa State Academy of Refrigeration, Odessa, Ukraine

Influenza virus A2/Hong Kong/1/68 was chosen to be the model object of the present study. The preparations with various haemagglutinating infectious activities (HIA) were under investigation. Chicken embryos were infected by this virus. After 48 hours the allantoic liquid was collected and new viral generation HIA was checked up through HA reaction. The virus-containing material was titrated on chorioallantoic membranes’ fragments and the infectious activity TID50 was evaluated. We have measured the fractal dimension (FD) of the virus containing preparations using the fractal diffraction patterns monitor (FDPM). The DP images’ frames were registered by digital VZM 1000 Color LabVideo system and processed with the use of original software FractImagePro elaborated in our group. It was demonstrated experimentally for the first time that FD is the reliable, objective and precise numerical parameter that describes the presence as well as HIA of a given viral material. The addition of anti-influenza immunoglobulin to the virus containing preparations has led to the significant changes of their FD. These changes, most probably, are due to the reaction of antigen-antibody type that takes place. The proposed FDPM method allows to detect the virus-cell interaction occurrence without any coloring and at the minimal virus-containing concentrations some minutes after the reaction has started. The application of FDPM method could be successfully performed even in the case of enveloped viral particles detection. We have shown experimentally for the first time that the FDPM method and device could be used for express diagnostics in drug design and clinical practice.

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COMBINED THE HUMAN IMMUNODEFICIENCY VIRUS AND NEUROSYPHILIS. ASPECTS OF TREATMENT OF THE LATTER

S.N. Potekayev, T.P. Bessarab, V.V. Belyaeva, E.V. Ruchkina, A.P. Kutsemilova, V.N. Dumkin, V.V. Pokrovsky
Russia Aids Center, Moscow, Russia

Now interaction between the human immunodeficiency virus (HIV) and Treponema pallidum (TP) is known. Such TP is more aggressive with enhanced multiplying.

Purpose: To summarize patterns of neurosyphilis (NS) in our HIV-positive patients (pts) in regard of the interaction mentioned above.

Methods: The diagnosis of HIV was based on ELISA and Western Blot, of NS – on Wassermann reaction with both antigens, FTA-ABS, TPI in the serum and CSF. (In the serum FTA-200, FTA-ABS, in the CSF FTA with undiluted CSF and Lange reaction were used.) Audiological investigations (audiometry, vestibulometry).

Results: Total 10 pts with HIV and NS were observed. All pts were homosexual males aged 24-41. CD4 number 450-50/mm³.
Two had asymptomatic NS (HIV signs: in 1 - GLP and perianal condylomata acuminata and in the other - acneform folliculitis of the back.). 1 pt had paralysis progressiva-like NS. (HIV: large-focus pityriasis versicolor of the thorax and of the neck. Audiologically: sensoneural deafness, moderate hearing loss.). Its peculiar signs were greatly reversible mind deficit and self-critics loss. 4 pts had tabes dorsalis (TD). (HIV: in 1 - cachexy, tbc and P. carinii pneumonia (AIDS), central vestibular syndrome; in 1 - polyneuropathy of the legs and of the feet, sensoneural deafness, mild hearing loss; in 1 - herpes simplex labialis, hyposmia; and 1 pt was asymptomatic HIV-carrier.). Its peculiarities were extremely severe ‘cold’ and ‘gunshot’ sensations in the legs and in the feet in 2 pts and prominent anisocoria in one. In the latter HIV-infection combined with TD progressed from asymptomatic to AIDS promptly. 2 pts had encephalitis. (HIV: in 1 - oral candidiasis; in 1 - recidiving herpes simplex labialis. Both had central vestibular syndrome, sensoneural deafness, mild hearing loss.). The encephalitis was distinguished with unusually severe dysarthria and headache. In 1 pt (lost to follow-up) cerebral gummas and meningoencephalitis were revealed as postmortem findings. (HIV: cachexy (AIDS)). High-dose of benzyl-PNC-natrii was a treatment of choice.

Conclusion: The interaction HIV-TP lead to unusual and more severe manifestations of NS in our pts. Otoneurological disorders were not uncommon findings. High-dose IV benzyl-PNC-natrii was quite effective in such NS.

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RECOMMENDATIONS FPR ANTIVIRAL TREATMENT AND PROPHYLAXIS OF INFLUENZA IN GERMANY

P. Wutzler, K.-D. Kossow, H. Lode, B.R. Ruf, H. Scholz, G.E. Vogel
Paul-Ehrlich Society for Chemotherapy, German Association for the Control of Virus Diseases, Germany

Antiviral drugs are a valuable supplementation to vaccines for the control and prevention of influenza. In Germany, amantadine, oseltamivir (Tamiflu®), and zanamivir (Relenza®) are approved. On behalf of the Paul-Ehrlich Society for Chemotherapy and the German Association for the Control of Virus Diseases, the first German Consensus Conference on the Antiviral Treatment and Prophylaxis of Influenza was held in June 2002. Based on available international and national data an expert panel of general practitioners, internists, paediatricians, gerontologists, epidemiologists, and virologists developed the following recommendations for the appropriate clinical use of the antiviral drugs: (1) Since oseltamivir (orally administered) and zanamivir (administered by inhalation) have apparently similar efficacy both drugs can be used alternatively. (2) Amantadine is not an alternative for the neuramindase inhibitors because it is not effective against influenza B viruses, it selects frequently resistant virus strains and it can cause adverse events of the central nervous system. (3) When influenza is prevalent in the community, patients with the clinical diagnosis of influenza should be treated if the symptoms are lasting not longer than 48 hours. Laboratory confirmation of influenza is necessary when influenza is not circulating in the community as well as in young children. (4) Although there are no data from clinical trials immuocompromised patients should also be treated when influenza has been diagnosed. (5) The prophylactic use of neuraminidase inhibitors can be recommended for persons with close contact to acutely ill persons.

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IMMUNOLOGICAL, EPIDEMIOLOGICAL AND ECONOMICAL EFFECTIVENESS OF ANNUAL INFLUENZA VACCINATION AMONG HEALTHY WORKING ADULTS

T.B. Spiridonova, S.G. Badogin, O.V. Demikhovska
Mechnikov Regional Hospital, Folk Medicine Institute, Dnipropetrovsk, Ukraine

Economic evaluation of seasonal influenza (FLU) and acute respiratory viral infections (ARVI) prevention still remains one of the most important points in public health, especially for healthy working adults in age group 18-65 years where the problem have been studying worst. Although the immunological and epidemiological aspects of annual FLU vaccination are known better, but some of them are also still discussible.

Methods: 49 462 workers of four large industrial enterprises were vaccinated annually during 4 years by influenza split-vaccine Fluarix (GSK). Non-vaccinated 1 831 workers of the same enterprises and 10 990 workers of two other similar enterprises formed internal and external control groups. Levels of serum antibodies against influenza were detected before, 7 and 30 days after vaccination. Coefficient of epidemiological efficacy (CEE) and efficacy index (EI) were calculated as a relation of FLU or/and ARVI morbidity levels in vaccinated and non-vaccinated groups. Evaluation of economic benefits was based on calculations of average illness duration, lost workdays and direct/indirect costs associated with vaccination against direct/indirect costs prevented by vaccination.

Results: 89-92% of vaccinated developed protective antibody level rapidly within
7 days after vaccination, 97-98% within 30 days. Fluarix vaccination demonstrated excellent epidemiological outcomes with CEE about 90% (84.8%-95.7%), EI from 7.1-8.5 up to 49.0 in season 2002/03 and extremely low level of adverse events (0.01-0.10% – active questioning). Significant economical benefits were one of the most important conclusions from the study: lost workdays level decreased by 59.25-79.35 for each 100 workers with economical effectiveness coefficient 9.08.