TABLE OF CONTENT

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1. EFFECT OF A FUNGIAL Cu/Zn SUPEROXIDE DISMUTASE ON THE FUNCTIONS OF PHAGOCYTIC CELLS IN YERSINIA ENTEROCOLITICA PYV (+) INFECTED MICE

E. Ivanova, D. Donkova, Z. Stefanova, R. Toshkova, M. Angelova, W. Voelter, H. Najdenski

ABSTRACT
Effect of Cu/Zn superoxide dismutase (SOD) isolated from the fungal strain Humicula lutea 103 (HLSOD) on the functions of phagocyte cells in Yersinia enterocolitica pYV(+) (YE) infected ICR mice and their survival rate was studied. Migration and killing activities of peritoneal macrophages, as well as phagocyte abilities of blood polymorphous-nuclear leukocytes (PMNs) and peritoneal macrophages were followed from days 3 to 21 post infection. Blood cultures and bacterial load of liver, spleen and small intestinies were examined too. A marked protective effect of HLSOD on the survivability of mice after lethal YE infection was established. The mortality percent (MP) in the control group of infected animals without treatment was 100% and strongly diminished (up to 17%) after treatment by 40U HLSOD. Acceleration of bacterial clearance from blood was another effect of HLSOD treatment. Augmented values of macrophage number in healthy and infected animals were established after HLSOD application with a maximum at day 15,h. Phagocyte indexes of macrophages from healthy animals increased under the action of enzyme treatment. The phagocyte number was 2 times increased in both groups of the infected animals. Macrophage killing indexes against S. aureus 209 were also increased in both groups of infected animals with a maximum at day 21s1 post infection. HLSOD restored the suppressed peroxide formation and killing ability of macrophages from infected animals against E. coli Ou>. HLSOD produced a marked stimulation effect on the migration ability of macrophages, better expressed in healthy animals. Additionally, HLSOD induced an increase of PMNs phagocyte indexes from the infected animals and the number of the in vitro engulfed S. aureus Smith cells was 1.5-2.0 times higher in both infected groups during the whole period of observation. The established protective effect of HLSOD on survivability of YE infected mice can be explained by the participation of this enzyme in the host oxidant-antioxidant balance, resulting in a restoration of the suppressed phagocyte, killing and migration abilities of peritoneal macrophages and blood PMNs of the infected mice.


2. COMPARISON OF THE SUSCEPTIBILITY TO 11 ANTIMICROBIAL AGENTS OF CLINICAL STRAINS BETA-HAEMOLYTIC GROUP A AND B STREPTOCOCCI ISOLATED FROM ADULTS AND REVIEW OF THE LITERATURE

B. Girgitzova, D. Chankova

ABSTRACT
Increasing antimicrobial resistance of group A and group B streptococci to some antibiotics has been observed during the last decade in Europe and worldwide. The aim of this study was to evaluate the susceptibility to antimicrobial agents of clinical strains group A and B streptococci isolated from adults and to compare our results to those from other parts of the world. Using the standard agar dilution method we studied the susceptibility to 11 antimicrobial agents of 156 S.pyogenes strains and 138 S.agalactiae strains isolated in 1999-2001. All isolates were susceptible to penicillin, cepha-lothin, cefuroxime and cefotaxime. The sensitivity to erythromycin in both groups was almost equal - about 96%, while clindamycin resistance (1,45%) was found only in group B streptococci. Resistance to chloramphenicol (3,62%), ciprofloxacin (1,45%), gentamicin (3,62%) and intermediate susceptibility to it (62,32%) was demonstrated again by group B streptococci. They also were resistant to tetracycline (92,76% against 9,62% for group A) and trimethoprim-sulfametoxazole (100% and 30,13% respectively). Among comparable studies concerning other countries our results demonstrate a bit lower or near the average values reported from Europe.


3. ISOLATION OF LEGIONELLAE FROM HOTEL WATER SUPPLIES IN SOFIA

E. Tsvetkova, I. Tomova, B. Halova, A. Tomov, V. Kassovsky, A. Mllanova

ABSTRACT
Investigations for the presence of legionella bacteria,in hotel water supplies in Sofia were performed. For this purpose one hundred and twenty five samples of potable water were collected from eleven hotels. A total of eighty-seven strains were isolated and determinate as Legionella spp. using culture, biochemical, iminuno-fluorescent (DFA) and gas-liquid chromotography methods. The legionella concentration ranged between 10Ч04 CFU/L. By the DFA technique the isolates were identified as Legionella pneiimophikt: Sg 1-2 strains, Sg 2- 5 strains. Sg3- 20 strains, Sg 4- 5 strains, Sg6-52 strains, Sg 8-2 strains. Sg 14- I strain. Twenty water samples from one of the hotels were detected for amoebae too. After cultivation and microscopic examinations eight strains of small free-living amoebae (FLA) were isolated, At the same hotel we traced out the water quality before and after decontamination by hyperchlorination. It was established that this procedure led to sharp decrease of the bacterial number or absence of culturable legionellae but after one-two months they appeared again. These data suggest that it is necessary to organize regular monitoring of hotel water supplies in Sofia for the presence and the concentration of legionellae.


4. CASE OF DIROFILARIASIS WITH MALE BRAEST LOCALIZATION CAUSED BY DIROFILARIA REPENS

I. Drandarska, D. Vuchev, M. Chergova

ABSTRACT
The rare case of dirofilariasis with male breast localization of Dirofilaria repens has been described. The parasitological diagnosis has been set up after diagnostically and therapeutically extirpation and histopathological examination of the tissue material.


5. ERYTHEMA MIGRANS - LIKE LESION ASSOCIATED WITH RICKETTSIA CONORI

R. Komitova, M. Murdjeva, E. Aleksandrov

ABSTRACT
Erythema migrans has been described as “pathognomonic” for early Lyme borreliosis. Mediterranean spotted fever, also transmitted by ticks, is a febrile illness with generalized rash, although few cases are “spotless”. Here we present a case with Erythema migrans-Hke lesion with serological evidence of Rickettsia conori and transitory anti Borrelia burgdorferi sero-positivity. A 58-year-old woman was admitted to the hospital in March 2000 with a 2-day history of low-grade fever and rash on her popliteal fossa. An engorged tick was removed from the same place, later identified as Demacentor marginatus. The rash resolved after 15 days of oral doxycycline. Three probable explanations have been discussed. Erythema migrans should not be considered pathognomonic for early Lyme borreliosis. To further establish Rickettsia conori as aethiologic agent of Erythema migrans-like lesion culture and PCR technique are strongly recommended.


6. APPLICATION OF RUBELLA IgG TOTAL ANTIBODY AVIDITY MEASUREMENT IN THE DIAGNOSIS OF RUBELLA INFECTION

Z. Mihneva

ABSTRACT
Objective: The measurement of total IgG antibody avidity in human sera is a useful technique in confirming the differentiation of an infection.

Materials and methods: Sera samples (n=79) from rubella immune patients (n=54),classified in four groups: I (group)-including cases (n=12) with clinically and serologically confirmed primary infection; ll(group)-including persons (n=25) with MMR vaccine in different post-immunization period; lll(group) - including women during pregnancy and women in vitro inseminate (n=15) and IV(group) - including cases (n=2) with rubella embriopathy (CRI) were investigated. ELISA test (commercial kit, firm Bio-Rad) - Platelia Rubella IgG was used. 8 molar urea in wash buffer as a protein denaturing agent was used too.

Results: Low avidity - antibody index (Al) below 30% (cut-off value) in a range between 10.80% to 25.54% was proved among cases with primary infection, immunized persons in the first three months following vaccination and in one of the rubella embryopathy cases. High avidity - antibody index (Al) above 30% was proved in convalescent sera (previous infection and immunization) - 68.90% (mean value).

Conclusion: The priorities of the IgG total antibody avidity measurement test as good diagnostical marker were discussed.


7. CLINICAL EXPERIENCE IN THE TREATMENT OF FLU PATIENTS WITH OSELTAMIVIR DURING THE INTERPANDEMIC PERIOD

V. Markovski, Z. Milenkovic, M. Dimzova, B. Tosevski

ABSTRACT
The influenza epidemics are very important health and economics issues. That explain why defining the methods for proper treatment and prophylaxis of influenza has become so important over the past few years. The study included people with present influenza symptoms throughout the last influenza season December 2001 - April 2002 confirmed by IIF, ELISA-IgM, isolation of the virus and with clear clinical symptoms. The goal of the study was on the basis randomized, double-blind study to evaluate the therapeutic effects of oselatamivir concerning the duration of the illness and symptoms. The study shows that the therapy with oselatinivir resulted in reduction of symptoms in febrile as well as in non febrile patients, smaller administration of symptomatic therapy, less complications as a secondary bacterial infections, lower administration of antibiotics. The drug oselatamivir is very well tolerated and only 5,9% of the treated patients noted intolerance with vomiting. From the present study the authors conclude that treatment of influenza with oselatamivir causes significant shortening of illness period as well as of clinical expression and symptoms of influenza.


8. INTRADERMAL TESTS WITH ALLERGENS S. AUREUS AND C. ALBICANS IN PATIENTS WITH ATOPIC ECZEMA/DERMATITIS SYNDROME

J. Kochetova, M. Mokronosova, B. Petrunov

ABSTRACT
Atopic eczema/dermatitis syndrome (AEDS) - is a chronic allergic skin disease. There is increasing evidence that infectious agents are important triggering factors in the pathogenesis of AEDS.

S. aureus has been studied as possible trigger factor. It is well known fact that the skin of 70-90% patients with AEDS is colonized with S. aureus in extremely high density (107 colony - forming units/cm2). In contrast only 5-10% of healthy individuals without any skin diseases are carriers S. aureus in low density of colonization. S. aureus is known as the producer of proteins with superantigenic properties: staphylococcal enterotoxin A (SEA), staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), toxic shock syndrome toxin -1 (TSST-1), staphylococcal exfoliative toxin (SET). At first, they can bind to the MHC II molecules of monocytes and dendritic cells and release a number of proinflammatory molecules such as interleukin-1 (IL-1) and tumor necrosis factor - alpha; at the second, they can stimulate the proliferation of T-cells with Vb chain expression . Moreover staphylococcal enterotoxins are considered as a new group of allergens. Specific IgE-antibodiesto SEA, SEB, SEC and TSST-1 have been detected in sera from 57% of patients. Basophils of the patients with AEDS were found to release histamine upon incubation with these toxins.

The relationship between the severity of AEDS and the density of S. aureus colonization in the skin was revealed. The moderate and severe courses of AEDS are characterized by the high density of staphylococcal colonization. Recently a number of studies have published that the severity of AEDS depends on presence of staphylococcal enterotoxins in the skin secreting by S. aureus.

The lipophilic yeast Malassezia spp is not only part of the normal human skin flora but also an opportunistic pathogen. Various recent reports have discussed Malassezia spp as a pathogenic factor in AEDS. C. albicans, unlike Malassezia spp., does not normally colonize the skin, but rather the gastrointestinal tract and the nasopharyngx. A pathogenic role for allergy of C. albicans has for long been proposed, particularly in asthma and rhinitis, and a high rate of positive immediate skin test to C. albicans has been observed in atopic patients. Allergenic cross reactivity between Malassezia spp. and C. albicans has been investigated and found to be significant.

The allergenic properties of Malassezia spp. and C. albicans are well known and few recombinant allergens have been obtained and characterized. The allergens have been recognized as mannoproteins. It was shown that predominantly sensitization to Malassezia spp. has been occurred in patients with head and neck skin lesions. C. albican as allergen, together with other antigenes (PPD, tetanic toxin, trichophytin), is as the marker of cell immunity and reveals delayed type hypersensitivity reactions in vivo. The aim of the study was to investigate immediate and delayed type skin reactivity by means of intradermal testing by allergens from S. aureus and C. albicans in patients with AEDS.

Atopic eczema/dermatitis syndrome (AEDS) - is a chronic allergic skin disease. There is increasing evidence that infectious agents are important triggering factors in the pathogenesis of AEDS.

S. aureus has been studied as possible trigger factor. It is well known fact that the skin of 70-90% patients with AEDS is colonized with S. aureus in extremely high density (107 colony - forming units/cm2). In contrast only 5-10% of healthy individuals without any skin diseases are carriers S. aureus in low density of colonization. S. aureus is known as the producer of proteins with superantigenic properties: staphylococcal enterotoxin A (SEA), staphylococcal enterotoxin B (SEB), staphylococcal enterotoxin C (SEC), toxic shock syndrome toxin -1 (TSST-1), staphylococcal exfoliative toxin (SET). At first, they can bind to the MHC II molecules of monocytes and dendritic cells and release a number of proinflammatory molecules such as interleukin-1 (IL-1) and tumor necrosis factor - alpha; at the second, they can stimulate the proliferation of T-cells with Vb chain expression. Moreover staphylococcal enterotoxins are considered as a new group of allergens. Specific IgE-antibodiesto SEA, SEB, SEC and TSST-1 have been detected in sera from 57% of patients. Basophils of the patients with AEDS were found to release histamine upon incubation with these toxins.

The relationship between the severity of AEDS and the density of S. aureus colonization in the skin was revealed. The moderate and severe courses of AEDS are characterized by the high density of staphylococcal colonization. Recently a number of studies have published that the severity of AEDS depends on presence of staphylococcal enterotoxins in the skin secreting by S. aureus.

The lipophilic yeast Malassezia spp is not only part of the normal human skin flora but also an opportunistic pathogen. Various recent reports have discussed Malassezia spp. as a pathogenic factor in AEDS. C. albicans, unlike Malassezia spp., does not normally colonize the skin, but rather the gastrointestinal tract and the nasopharyngx. A pathogenic role for allergy of C. albicans has for long been proposed, particularly in asthma and rhinitis, and a high rate of positive immediate skin test to C. albicans has been observed in atopic patients. Allergenic cross reactivity between Malassezia spp. and C. albicans has been investigated and found to be significant.

The allergenic properties of Malassezia spp. and C. albicans are well known and few recombinant allergens have been obtained and characterized. The allergens have been recognized as mannoproteins. It was shown that predominantly sensitization to Malassezia spp. has been occurred in patients with head and neck skin lesions. C. albican as allergen, together with other antigenes (PPD, tetanic toxin, trichophytin), is as the marker of cell immunity and reveals delayed type hypersensitivity reactions in vivo. The aim of the study was to investigate immediate and delayed type skin reactivity by means of intradermal testing by allergens from S. aureus and C. albicans in patients with AEDS.


9. SEASONAL DYNAMIC OF MITE ALLERGENS EXPOSURE (DER 1, DER 2) AND HOUSE DUST MITES IN MOSCOW'S APARTMENTS

T. Zheltikova, V. Gervajieva, M. Mokronosova

ABSTRACT
It is well known that the number of pyroglyphid mites and, therefore, exposure to mite allergens varies during the year. Seasonal dynamic of mites has its specific geographical features, which plays a causal role in the development of bronchial hyperreactivity in patients with asthma. In this connection the monitoring of house dust mites and mite allergen levels in patients’ homes should be considered as the strategy for the treatment of bronchial asthma. Our aim was the study of the seasonal dynamic house dust mites and mite allergens exposure (Der 1, Der 2) in some asthma patients’ apartments in Moscow. In 1994-1996 the number of pyroglyphid mites and exposition to mite allergens of groups 1 and 2 were monitored in the apartments of asthma patients sensitized to house dust mites. The study was carried out in 5 apartments, random chosen as models, by means of the acarological analysis and ELISA with the use of monoclonal antibodies to mite allergens. The data obtained in this study made it possible to determine the periods of rises and falls in the concentration of mite allergens in the dust of apartments in Moscow. The seasonal dynamics of exposition to mite allergens of groups 1 and 2 had a regional character. One period of a rise in the concentration of mite allergens (>2 pg/g of dust), common to all model apartments under study in Moscow, was established: the end of August -the beginning of October. Ail other periods of rises in the concentration of allergens were specific for each individual apartment. Thus, the prognostication of the periods of rises in the concentration of allergens in a patient’s environment proved to be rather difficult. Therefore, prior to the prescription of specific immunotherapy to an individual patient exposition to mite allergens should be established and, with such concentration exceeding 2 pg/g of dust (100 mites/g of dust), the intensive sanitary and hygienic treatment of the apartment should be carried out.


10. STUDY OF THE FC FUNCTION OF THE IMMUNOGLOBULIN IN IMMUNOVENIN-INTACT

J. Natcheva, Z. Mihneva, A. Mihailov, I. Bineva

ABSTRACT
Immunovenin-intact (immunoglobulin for intravenous use) contains immunoglobulin in its native form and is presented as ready to use liquid. The recommendation of the European Pharmacopoeia to prove the nativity of the IgG molecules (their Fc function) by the assessment of the level of rubella antibodies was followed. The measurements of these antibodies during the shelf life of the preparation showed that their levels were preserved and this gave assurance of the efficacy and stability of Immunovenin-intact. The method of rubella radial haemolysis was applied to prove that the Fc function of the immunoglobulin in this preparation was not impaired.


11. COMPARATIVE CHARACTERIZATION OF ANTIGENS ISOLATED FROM TOXOCARA CANIS

I. Rainova

ABSTRACT
Toxocarosis is parasitic disease in humans, caused by the larval form of dog nematode Toxocara canis. For diagnosis of the disease different serological tests are used which demonstrate the presence of specific antibodies in serum specimens of suspected patients. In the development of the immunological methods two kinds of antigens are used mainly - somatic from adult forms and excretory-secretory (metabolic) from parasite larvae cultivated in vitro. The aim of the presence work is to make comparative characterization of these two antigens using SDS-electrophore-sis and to study their specificity in enzyme -linked immunosorbent assay (ELISA). The results showed that by electrophoretic separation in SDS-PAGE the two kinds of antigens differ in number of isolated fractions and their molecule mass in kDa. The testing in ELISA with sera from patients, reacted negative for toxocarosis in another tests and from patients with other parasitic diseases, showed that E/S antigen isolated from T.canis larvae is more useful and possess enough specificity as a diagnostic reagent.


12. EXSTRADIGESTIVE DISEASES ASSOCIATED WITH H.PYLORI INFECTION

I. Mladenova-Hristova

ABSTRACT
H. pylori gastric infection is a chronic process lasting for decades. The persistent infection induces a chronic inflammatory and immune response able to induce lesions both locally and remote to the primary site of infection. H. pylori infection may be a risk factor for the development of coronary heart disease. In a recent cross-sectional study Mendall et al, confirmed that CHD is associated with C reactive protein and with high serum levels of IL-6 and TI\IF-a demonstrated predictors of myocardial infarction and stroke. These cytokines are associated with high levels of fibrinogen, sialic acid, triglycerides and reduced HDL-cholesterol. Following H. pylori infection, a link, between increased levels of cytokines and an increased level of C reactive protein, leading to CHD, can be hypothesized. A significant amelioration of both - Raynaud phenomenon and migraine symptoms has been described after H. pylori eradication. The prevalence of CagA(+)H. pylori strain was higher in infected subjects with autoimmune thyreoiditis than in infected controls. An association with chronic idiopathic urticaria has been proposed with the pathogenetic mechanism related to an increase in gastric vascular permeability during H. pylori infection, resulting in a greater exposure of the host to alimentary allergens.


13. TRANSMISSION OF HELICOBACTER PYLORI: EVIDENCE FOR FECAL-ORAL ROUTE

I. Mladenova-Hristova

ABSTRACT
The evidence for the fecal-oral route of transmission of H. pylori is still controversial. H. pylori DNA has been detected in feces of infected subjects by some researchers but not others. It has been demonstrated that H. pylori can live for several days in milk and tap water in its infectious bacillary form and in river water for several months in a coccoid form. Transmission has been linked to consumption of vegetables grown in regions where human feces is used as fertilizer. Although the principal reservoir for H.pylori infection appears to be people, H.pylori has been isolated from nonhumans primates and domestic cats. The most recent reservoir suggested for H.pylori transmission is the housefly. However, evidence is lacking that H.pylori can be isolated from flies that have been in contact with H.pylori infected feces and that H.pylori can be transmitted from contaminated flies to food in a quantity sufficient to infect humans. In fact, nosocomial transmission of H.pylori is the only proven mode of transmission. Understanding the route(s) by which H.pylori infection spreads is fundamental for development of strategies for the infection’s control.


 Editorial Board
Editor-in-Chief
Prof. B. Petrunov, MD, DSc

Executive Editor
Prof. H. Taskov, MD, DSc
Assoc. Prof. R. Kurdova, MD, PhD
Assoc. Prof. R. Kotzeva, MD, PhD
Assoc. Prof. N. Gatcheva, MD, PhD
Assoc. Prof. T. Kantardjiev, MD, PhD
Prof. P. Nenkov, MD, DSc

Production Manager
Dr. V. Gatcheva

Technical Assistance
T. Koshev

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