TABLE OF CONTENT

dl

1. ENZYME - LINKED IMMUNOSORBENT ASSAY FOR ESTIMATION OF IMMUNOGLOBULIN G ANTIBODIES AGAINST PERTUSSIS IN HUMAN SERA

R. Alexiev, K. Hadjiisky and PI. Nenkov

ABSTRACT
Estimation of pertussis antibody in human sera is very important for investigation of efficacy of vaccination against whooping cough. An enzyme-linked immunosorbent assay was development for measuring immunoglobolin G antibodies against pertussis in human sera. The assay was done in plastic plates coated with bacterial cells. The bacterial agglutination test is time consuming, thus challenging us to develop more rapid and simple alternative in vitro test. Simple to perform, economical and precise, the ELISA seems to be a very practical procedure for seroepidemiological purposes.


2. EXPRESSION OF PERFORIN AND GRANZYME B IN TOTAL CD8 T LYMPHOCYTES OF PATIENTS WITH CHRONIC HIV AND EBV INFECTIONS.

M. Muhtarova, M. Nikolova, S. Magaev, M. Petrova, K. Kostov, I. Elenkov, H. Taskov

ABSTRACT
The mechanisms affecting CD8 T cell functions in the course of chronic viral infections and their phenotypic equivalents are a matter of intensive research. Together, granzyme B (GzmB) and perforin (Per) are essential mediators of cytotoxicity. Using flow cytometry, we studied GzmB/Per co-expression in total CD8 T cells of subjects with chronic untreated HIV-1 or EBV infection, in relation to CD8 T cell differentiation subsets defined by CD160/ CD27/CD28 expression and the Treg subset CD4+CD25high. Further on, we looked for association between the patterns of GzmB/ Per expression and markers of disease progression. GzmB+Per+ and GzmB+Per- CD8 T cells were increased in HIV and EBV patients, but only the single-positive subset was significantly increased in both patients groups as compared to controls (mean 41 % and 11 % vs. 5%). Moreover, the GzmB+Per - CD8 subset was disproportionately increased in HIV infection, differing significantly from both EBV (p<0.01) and control (p<0.001) groups. The level of GzmB+Per- CD8 T cells in HIV patients correlated with the level of antigen-primed CD28- CD8 T cells lacking CD160 expression (R= 0.6, p<0.001). While the discordant GzmB/Per expression in HIV patients did not correlate with CD4 AC or CD38ABC as a marker of viral replication, a positive correlation was established with the percentage of CD4+CD25W9h Treg cells. In conclusion, the downregulation of CD160 on CD8 T cells in the course of chronic HIV infection may be a significant correlate of impaired cytotoxic functions, at least in part related to CD4+ Treg mediated suppression.


3. CLINICAL CASE OF TRICHINELLOSIS IN A SPLEENECTOMIZED PATIENT

D. Vuchev, M. Vretenarska, M. Ivanova, D. Nikolova, R. Georgiev

ABSTRACT
A prolonged course of moderately severe form of trichinellosis was observed in a patient, with previous spleen and kidney ectomy. After etiological treatment with Zentel and prolonged pathogenic therapy, the patient went trough a half-year dispensary observation, until recovery. Peculiarities observed in clinical and laboratory constellations were described.


4. ANTICESTODAL ACTIVITY OF NEW BENZIMIDAZOLE DERIVATIVES

I. Marinova-Hamova, K. Anichina, D. Vutchev, A. Mavrova

ABSTRACT
Two chemicals containing a condensed thiazole nucleus in their structure, are investigated in the present study. The compounds are tested in vitro on isolated scolices of Echinococcus granulosus in concentrations from 5 to 320 mg/ml and in vivo on laboratory animals - white mice infected with Hymenolepis папа in dosage 20 mg/ml. One of the benzimidazoles, shown prominent activity is subject to additional investigations.


5. STOOL ANTIGEN TEST FOR DIAGNOSIS OF HELICOBACTER PYLORI INFECTION

G. Zhelezova, L Boyanova

ABSTRACT
Helicobacter pylori is now generally accepted as the main etiological agent in chronic active gastritis and peptic ulce and risk factor for the development of gastric cancer. The two major categories of diagnostic methods for diagnosis are invasive tests, or those that require endoscopy, and noninvasive, or nonendoscopic tests. The aim of this study was to systematically review the experience on H. pylori stool immuno assay antigen tests as a relatively new noninvasive diagnostic test. The review was intended to evaluate the usefulness of different stool antigen tests in the pretreatment diagnosis of H. pylori infection and after eradication therapy; in adult and in pediatric patients; and factors which affect the accuracy of the stool antigen tests.


6. FOUR CASES OF IMPORTED BRUCELLOSIS IN BULGARIA

O. Boykinova, I. Stoilova, I. Baltadjiev, S. Novakov, V. Nikolov, M. Troyancheva, Ts. Kazakova

ABSTRACT
Brucellosis is a typical zoonosis infecting domestic animals and frequently humans occupationally linked to them. Isolated cases of imported infection have been observed in Bulgaria though this disease was considered to be eliminated since 1974. The social and economic changes and migration of the population in the last decade make brucellosis current issue in our country. The aim of our study is to make a clinical and epidemiological surveillance of four cases of imported brucellosis from Greece and to display the risk of employment in enzootic regions. The patients are with manifested brucellosis appearing as systemic infection with multiorgan location. The diagnosis was proved by complex epidemiological and routine clinical, laboratory and serological methods. In conclusion familiarity with manifestations of brucellosis, the optimal laboratory techniques for its diagnosis and preliminary health education for the people going to work in risk endemic regions could help physicians protect the public against this reemerging, under-recognized zoonosis.


7. PRELIMINARY CHARACTERIZATION OF THE RESISTANCE MECHANISMS IN CARBAPENEM-RESISTANT CLINICAL ISOLATES OF PSEUDOMONAS AERUGINOSA

T. Strateva, V. Ouzounova-Raykova, B. Markova, Y. Marteva-Proevska, I. Mitov

ABSTRACT
A total of 59 clinical isolates of Pseudomonas aeruginosa resistant to all beta-lactams, including to carbapenems, were collected during 2001-2005 from 5 university hospitals in Sofia to assess the current levels of antimicrobial susceptibilities and to preliminary evaluate the resistance mechanisms. Antimicrobial susceptibilities were detected by a disk diffusion method and Etest (AB Biodisk). Polymerase chain reaction amplification and sequencing of blaIMP-Iike, blaVIM-like, and blaVEB-1, were performed. The antibiotic resistance rates were: to ceftazidime - 100 %, cefepime - 100%, imipenem - 100%, meropenem - 100%, amikacin- 86.4%, gentamicin - 86.4%, ciprofloxacin - 81.4%. IMP-and VIM-type metallo-beta-lactamases (MBLs) from molecular class B were not detected. Structural genes for VEB-1 extended-spectrum beta-lactamases (ESBLs) were found in 34 (57.6 %) of the isolates. In conclusion, the resistance to all beta-lactams, including toward carbapenems, in the studied strains of P. aeruginosa was not related to enzymatic hydrolysis by MBLs. It was resulted from the combination of different mechanisms such as: OprD deficiency, ESBLs production, derepression of AmpC cephalosporinase and overexpression of active efflux systems. VEB-1 appears to have a significant presence among carbapenem-resistant P. aeruginosa isolates from Sofia.


8. OCCURRENCE OF CTX-M-3 EXTENDED SPECTRUM BETA-LACTAMASE PRODUCING SALMONELLA BRANDENBURG IN BULGARIA

P. Petrov, G. Asseva, T. Kantardjiev, P. Padeshky, N. Vlahova, I. Ivanov, A Ivanov, M. Nikolova

ABSTRACT
The aim
of the present work was to study the genetic profile and resistance to antibiotics of a rare serotype of Salmonella causing a cluster of cases among infants and children in Bulgaria.

Materials and methods: nine strains of Salmonella Brandenburg were isolated from infants and children with acute enterocolitis in the beginning of 2005. Culture, identification, serotyping and antimicrobial susceptibility testing to fourteen antibiotics have been performed. PCR with primers detecting bla-CTX-M genes and PFGE after restriction with Xbal were applied.

Results: A single PFGE type of strains confirmed the occurrence of a cluster of cases caused by Salmonella Brandenburg among infants and children. The strain was found to be multidrug- resistant. It produced a CTX-M- 3 extended- spectrum beta-lactamase.

Conclusion: Our findings proved the etiological role of a unique genetic variant of Salmonella Brandenburg in a cluster of cases among infants and children. This is the first report for Bulgaria of Salmonella Brandenburg producing a CTX-M-3 extended-spectrum beta-lactamase.


9. HAPTOGLOBIN AS A MARKER FOR LIVER INJURY IN HEROIN ADDICTS

J. Radenkova-Saeva, R. Kostadinova, S. Marinova, B. Petrunov

ABSTRACT
The aim of the study is to evaluate the level of haptoglobin as a part of immune reactivity in heroin addicts. In the study are included 35 heroin abusers between 16 and 43 years old. The level of transaminases (ASAT and ALAT) have been investigated and the quantity of haptoglobin in the patient sera have been determined by Manchini’s radial diffusion test in agar/agarose gel using immunoplates by “Immunotest”, Sofia. In 15 (43%) of the studied patients we found statistically significant elevated level of transaminases above the referent values. The haptoglobin was also significantly increased (p<0,05) in 10 (28,8%) of heroin abusers. Having in mind the normal level of haptoglobin in humans -0,3 to 2g/l - we found in our studied patients its value between 1,58 - 4,8 g/l what suggests that heroin may disturb the liver function provoking increased level of transaminases and haptoglobin. In conclusion we may say that haptoglobin could be used as a marker for liver disorders in heroin addicts.


10. DIAGNOSTIC STUDIES ON THE ETIOLOGICAL ROLE OF RESPIRATORY SYNCYTIAL VIRUS AND INFLUENZA VIRUSES IN HOSPITALIZED CHILDREN

S. Pavlova, T. Hadziolova, R. Kotzeva

ABSTRACT
Respiratory syncytial virus (RSV) role as causative agent of severe respiratory system diseases in young children, is a priority problem world-wide. Influenza viruses annual epidemic incidence also has an impact over the morbidity rate in this age group. In the period October 2005 - April 2006 in the Laboratory of Influenza and Acute Respiratory Diseases (ARD) comparative studies have been conducted for RSV and influenza viruses on 359 samples as a total originating from hospitalized children up to the age of 5. There is no parallelism between the virologic and serologic investigations in one and the same children. In the virologic investigation [isolation, rapid immunosorbent tests and immunofluorescence assay (IFA)] of 174 samples 14 (8,04%) have proved RSV positive, and 10 (5,7%) have been positive for influenza viruses type A. In the EUSA IgG testings of 185 single serum pairs, 42 (22,7%) RSV positive, and 52 (28,1%) influenza viruses type A positive results have been obtained. The results obtained confirm the reference literature data for the RSV and influenza viruses concurrent circulation among the age group indicated. The present survey enrich the diagnostic studies as regards the etiological role of these viruses in childhood by contemporary data, and promotes (initiates) competent collaboration between virologists and pediatricians.


11. USE OF NEURAMINIDASE INHIBITORS AND RESISTANCE IN PERSPECTIVE

P. Genova, T. Hadzhiolova, S. Pavlova, D. Dundarova, R. Kotseva

ABSTRACT
Of the four antiviral drugs currently available for the treatment of influenza A infection (two neuraminidase inhibitors and two M2 ion channel inhibitors), only the neuraminidase inhibitors osettamivir and zanamivir are also active against influenza B. All drugs are most effective if started within a few hours of the onset of symptoms and are generally licensed for use within 48 hours of the first symptoms. The neuraminidase inhibitors, oseltamivir and zanamivir, have fewer side effects than the M2 ion channel inhibitors rimantadine and amantadine, and drug resistance seems to develop less frequently. Numerous treatment studies in healthy adults have shown that neuraminidase inhibitors, when taken within 36 to 48 hours after the onset of symptoms, decrease the symptomatic illness by one or two days (Hayden 1997, Monto 1999, Treanor 2000, Cooper 2003, Whitley 2001, Aoki 2003). Early initiation of treatment is decisive for treatment efficacy (Aoki 2003, Kawai 2005). When started within the first 12 hours following the onset of fever, neuraminidase inhibitors shortened the illness by more than three days, in comparison to treatment that was started at 48 hours. The duration of fever, severity of symptoms, and time to return to normal activity also correlated with the time of initiation of antiviral intervention. The introduction of neuraminidase inhibitors was an important step for the more efficient control of human influenza infection. Today, neuraminidase inhibitors are the only drugs effective against recently isolated highly pathogenic avian influenza viruses in humans.


12. ANTIMICROBIAL SUSCEPTIBILITY OF URINE PATHOGENES ISOLATED FROM PATIENTS WITH BENIGN PROSTATIC HYPERPLASIA TREATED WITH UROSTIM

V. Grigorova, S. Stratev, F. Shargabi, N. Kolev, P. Panayotov, R. Kostov, O. Mihaylov, Ts. Georgiev, V. Dunev, B. Atanasov

ABSTRACT
Benign prostatic hyperplasia (BPH) is one of the most frequent reasons for recurring infections of the urinary tract. To determine the antibiotic susceptibility of microorganisms isolated from urine of patients with BPH and the results of immunotherapy with urostim. A total number of 126 urine samples of patients with BPH treated in the Clinic of Urology during 2005, were examined. Isolated strains were identified by conventional methods. E. coli and K. pneumoniae strains were screened for ESBLs production. Susceptibility to antibiotics was determined by using disc diffusion method of Bauer-Kirby, according to NCCLS. Immunotherapy with urostim for a 3-month period was applied to 36 patients and the urine was monitored on a monthly basis. The patients were divided into two groups: Group 1-12 patients with asymptomatic bacteriuria treated with urostim only, and Group II - 24 patients with symptomatic bacteriuria treated with urostim and antibiotic. Fifty strains were isolated from 40 patients with significant bacteriuria and pyuria. The most frequently isolated microorganisms were: E. coli (42%), K. pneumoniae (18%) and E. faecalis (12%). Producers of ESBLs among E. coli strains were 61.9%, and among K. pneumoniae - 77.8%. The E. coli and K. pneumoniae strains remained susceptible to imipenem and meropenem. The E. faecalis strains were susceptible to vancomycin and teicoplanin. After conducted treatment liquidation of the uroinfection were registered in 10 (83.3%) patients from Group I and in 18 (75.0%) patients from Group II. The bacteriuria remained persistent in 2 (16.7%) patients from Group I and in 6 (25%) patients from Group II. The immunotherapy with urostim does not exclude treatment with antibiotics in order to achieve highest therapeutic results.


13. A RAPID METHOD FOR DIAGNOSIS OF RARE FORMS OF TULAREMIA - ANALYSIS OF SWABS

P. Padeshki, I. Ivanov, M. Mincheff, S. Nikolaeva, T. Kantardjiev

ABSTRACT
The current report describes some of our findings during the tularemia outbreak 1997-2005 in Bulgaria. 285 cases were serologically and clinically confirmed for tularemia. Herein we describe the laboratory findings and the diagnosis of oculoglandular forms of infection. Oculoglandular tularemia was diagnosed in four patients by culture, immunofluorescent antibody analysis and polymerase chain reaction assay. Histological methods were applied for characterization of ocular tularemia granuloma. Three F. tularensis strains were isolated and characterized. One strain was isolated from an ocular swab specimen obtained from a seronegative patient. We report for the first time a successful application of diagnostic PCR performed directly on ocular swab specimen. We also describe the histological picture of a conjunctival granuloma in the course of infection. The proposed strategies are not invasive and could represent a new approach for resolving rare and hard-to-diagnose oculoglandular forms of tularemia.


14. SFG - RICKETTSIOSES IN BULGARIA

E. Alexandrov, A. Kunchev, N. Nedelchev, D. Alexandrova, B. Kamarinchev, S. Martinov, R. Komitova, R. Filipova, T. Kamenova, Z. Dimitrova

ABSTRACT
One hundred years ago, H.T. Ricketts laid the foundations of the research of tick spotted fevers (SFG-rickettsioses). The Mediterranean (Marseilles) spotted fever (MSF) was recognized and described for the first time in Bulgaria in 1948. We would like to report our studies and experience by dedicating them to H.T. Ricketts. From 1948 to 1970 (wave I period covering 22 years), a total of 240 cases had been recorded as sporadic cases or small outbreaks. Wave II of MSF began in 1993 by a sharp raise of the incidence. Until 2003, the number of cases was 20-25 times higher compared to the entire wave I 22-year period. 1,000-1,500 cases were recorded annually. The studies clarified that the key factors determining MSF case occurrence in Bulgaria were three: the stray population; the regular deacarization of farm animals; and the way of their breeding - concentrated in large animal breeding farms or in multiple small farms. Currently, a National Program for Combating and Prevention of Tick-borne Infections is being developed in Bulgaria, which is using also the experience accumulated in the MSF studies.


 

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

Production Manager
Theodor Koshev

Editorial Board
Prof. H. Taskov, MD, DSc
Prof. P. Nenkov, 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

Please publish modules in offcanvas position.

This site uses COOKIES to ensure optimal user experience.