Introduction: In surgical procedures, tissue damage results in the release of a number of bioactive substances. Calcitonin gene-related peptide (CGRP) is a peptide released from sensory nerves, which determines its role in pain sensation. Its distribution in tissues deter­mines its role as a primary afferent neurotransmitter. Aim: To determine the effect of CGRP on postoperative pain and reactive inflammatory process after surgical removal of impacted mandibular third molars, as well as the factors that have influence upon the perception of pain. Materials and methods: Forty patients with bilaterally impacted mandibular third molars were included in the study. Venous blood samples were collected before and 24 hours after the surgical procedure in order to test their serum levels of CGRP and procalcitonin. Two weeks later the procedure was repeated. The difficulty of the surgical procedure, its duration and complications were assessed in all patients. Results: The influence of some of the studied factors upon postoperative pain was established. Differences in the sensation of pain between the two sexes were found when comparing pain intensity reported by the patients. Significant difference between pain inten­sity after the 1st and 2nd surgical procedures (6 hours) was found in females (Z=2.63, p=0.009;), whereas in males the difference was observed at 24 hours (Z=1.99; p=0.047). Regarding the existence of sex-related association, а significant, strong positive correlation between CGRP levels after the 1st and 2nd surgical procedures (24 hours) was found in males (rxy=0.78; p=0.004), whereas in females this correlation was also significant, although moderately significant (rxy=0.44; p=0.020). CGRP levels at the first and second extractions were generally similar in males, and not as much in females. We proved significantly moderate positive association between CGRP and pulse levels measured before the second surgery (rxy=0.37, p=0.021). Conclusion: The results of our study suggest a significant role of CGRP in reactive (neurogenic) inflammation.

AIM: The aim of this study was to introduce a micromeritic procedure (a statistical approach for small objects) in indirect immunofl uorescence assay (IFA) to fi nd objective quantitative parameters of antinuclear antibody (ANA) patterns which could support a diagnosis of auto-immune diseases. MATERIALS AND METHODS: Sera of patients with systemic autoimmune diseases, McCoy-Plovdiv serum-free cell line, goat anti-human immunoglobulin-G FITC-conjugate, fl uorescent microscope, computer-assisted digital image processing, analysis using a micromeritic procedure, ANOVA. RESULTS: Three ANA fl uorescent patterns (homogeneous, rim and speckled) were analyzed by the micromeritic procedure. Parameters for the image brightness of the pixels (pixel grey value) were obtained and discussed as objective characteristics of fl uorescent patterns: maximum ANA-linkage volume and surface density were established for the objects with speckled localization pattern. CONCLUSION: The micromeritic method for getting objective quantitative values of ANA fl uorescent patterns in indirect immunofl uorescent assay might be a valuable tool aiding in immunological diagnosing if integrated in a laboratory software package. РЕЗЮМЕ ЦЕЛЬ: Работа ставит себе целью ввести микрометрическую процедуру (статистический анализ, применя-емый для небольших объектов) в индиректный флюоресцентный анализ в целях получения объективных количественных параметров при анализе локализации антинуклеарных антител в диагностике аутоим-мунных заболеваний. МАТЕРИАЛЫ И МЕТОДЫ: Применены: сыворотки пациентов с системными аутоим-мунными заболяваниями; безсывороточная клеточная линия McCoy-Plovdiv; маркированный флюорохро-мом FITC иммуноглобулин; флюоресцентный микроскоп; цифровизирование и обработка изображений; микрометрический анализ; ANOVA статистическая обработка данных. РЕЗУЛЬТАТЫ: Три вида локализации антиген-антитела (ANA) реакции -гомогенная, периферическая и петнистая -анализированы микроме-трическим методом. Считается, что яркость пикселов флюоресцентных изображений служит объектив-ной характеристикой вида локализации. Максимальный объем и поверхностная плотность связанных с субстратом ANA получены для петнистого типа локализации. ЗАКЛЮЧЕНИЕ: Mикрометрический метод количественной характеристики типа ANA локализации в индиректной иммунофлюоресценции может оказаться ценным дополнительным инструментом для анализа в иммунологических диагностических ла-бораториях с возможностью для включения в пакет программного обепечения (sofware) компьютерной программы для обработки изображений.Ключевые слова: антинуклеарные антитела, иммунофлюоресценция, цифровой анализ изображений, FITC, распределение размера пикселя lens -40x. The reaction between ANA and nuclear antigens of McCoy-Plovdiv cells was visualized as apple-green staining of nuclear structures under UV light. The intensity and patterns of fl uorescence were assessed and recorded.Selection of fl uorescent patterns for analysis. Three nuclear fl uorescent patterns, corresponding to different ANA localization in the nucleu...

Objective. To study both the molecular discrimination of D-tartrate fermenting and non-fermenting strains of Salmonella enterica subsp. enterica serotype Paratyphi B isolated from patients with paratyphoid fever and the clinical course of this disease. Materials and methods. The isolates examined were from children aged 3 months to 9 years. A total of 33 Salmonella strains were serotyped as Salmonella Paratyphi B, with an antigenic formula based on O- and H- antigens: 1,4, [5], 12: b: 1,2 by Kauffmann–White classification. Results. Multiplex PCR analysis confirmed all tested strains as d-tartrate fermenting (dT+), also referred to as variant Java. Discussion. We found that the most common cause of paratyphoid fever among children in Bulgaria is variant Java Salmonella Paratyphi B. Most children had classic symptoms of acute gastroenteritis – fever, watery diarrhea and vomiting.

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Въведение: Неферментиращите бактерии с клинично значение Acinetobacter baumannii и Pseudomonas aeruginosa, обичайно считани за опортюнистични патогени, днес заплашват да се превърнат в едни от най-проблемните причинители на инфекции. Терапевтичните рестрикции от една страна се дължат на тяхната вродена резистентност към широк набор от антибиотици, а от друга -на все по-нарастващата придобита резистентност. Цел: Да се анализират най-честите механизми на карбапенемна резистентност сред неферментиращите бактерии в страната и да се препоръчат опции за справяне с терапевтичните рестрикции. Методи: Обикновено се използват както фенотипни, така и генетични методи за определяне на резистентност към карбапенеми. Най-чести фенотипни методи за детекция на карбапенемази при неферментиращите бактерии са Модифицираният Ходж тест и Carba NP-тестът. Класически PCR се използва за доказване на гени, кодиращи карбапенемази, а qRT-PCR -за оценка на експресията на ключови ефлукс помпи, вродената AmpC и трансмембранния OprD-рецептор при Pseudomonas. Резултати и обсъждане: Две независими проучвания в България доказаха, че свръхекспресията на MexXY-OprM ефлукс помпата е главна при- ABSTRACT Introduction:The non-fermenters of clinical significance Acinetobacter baumannii and Pseudomonas aeruginosa, usually considered as opportunistic pathogens, nowadays emerge to evolve in some of the most problematic causative agents of infections. The therapeutic restrictions are determined on one hand due to their intrinsic resistance to a great variety of antibiotics and on the other, due to the arising acquired resistance.Aim: The aim of this article is to analyze the most often detected mechanisms of carbapenem resistance among non-fermenters in the country and to recommend options to manage the therapeutic restrictions.Methods: Both methods -phenotypic and genetic, are usually combined to evaluate the mechanisms of resistance to carbapenems. As phenotypic tests for detection of common carbapenemases a Modified Hodge test and a Carba NP-test are used. A classical PCR investigation is used to detect the most frequent carbapenemases for both microorganisms and a qRT-PCR to assess the expression of main efflux pumps, the intrinsic AmpC and the transmembrane receptor OprD in Pseudomonas.Results and Discussion: Two independent studies proved that in Bulgarian clinical isolates P. aeruginosa, the increased expression or overexpression of MexXY-OprM efflux pump, is the main mechanism of determining carbapenem resistance. The main reason for carbapenem resistance in A. baumannii iso-