E. Novakova

Pneumococcal capsular polysaccharides conjugated to protein D for prevention of acute otitis media caused by both Streptococcus pneumoniae and non-typable Haemophilus influenzae: a randomised double-blind efficacy study

Summary Background Acute otitis media is one of the most commonly-diagnosed childhood infections. This study assessed the efficacy of a novel vaccine that contained polysaccharides from 11 different Streptococcus pneumoniae serotypes each conjugated to Haemophilus influenzae -derived protein D in prevention of acute otitis media. Methods 4968 infants were randomly assigned to receive either pneumococcal protein D conjugate or hepatitis A vaccine at the ages of 3, 4, 5, and 12–15 months and were followed-up until the end of the second year of life. Middle-ear fluid was obtained for bacteriological culture and serotyping in children who presented with abnormal tympanic membrane or presence of middle-ear effusion, plus two predefined clinical symptoms. The primary endpoint was protective efficacy against the first episode of acute otitis media caused by vaccine pneumococcal serotypes. Analysis was per protocol. Findings From 2 weeks after the third dose to 24–27 months of age, 333 clinical episodes of acute otitis media were recorded in the protein D conjugate group (n=2455) and 499 in the control group (n=2452), giving a significant (33·6% [95% CI 20·8–44·3]) reduction in the overall incidence of acute otitis media. Vaccine efficacy was shown for episodes of acute otitis media caused by pneumococcal vaccine serotypes (52·6% [35·0–65·5] for the first episode and 57·6% [41·4–69·3] for any episode). Efficacy was also shown against episodes of acute otitis media caused by non-typable H influenzae (35·3% [1·8–57·4]). The vaccine reduced frequency of infection from vaccine-related cross-reactive pneumococcal serotypes by 65·5%, but did not significantly change the number of episodes caused by other non-vaccine serotypes. Interpretation These results confirm that using the H influenzae -derived protein D as a carrier protein for pneumococcal polysaccharides not only allowed protection against pneumococcal otitis, but also against acute otitis media due to non-typable H influenzae . Whether this approach would also allow improved protection against lower respiratory tract infections warrants further investigation.

Costs analysis of the treatment of imported malaria

BackgroundTo document the status of imported malaria infections and estimate the costs of treating of patients hospitalized with the diagnosis of imported malaria in the Slovak Republic during 2003 to 2008.Case studyCalculating and comparing the direct and indirect costs of treatment of patients diagnosed with imported malaria (ICD-10: B50 - B54) who used and not used chemoprophylaxis. The target sample included 19 patients diagnosed with imported malaria from 2003 to 2008, with 11 whose treatment did not include chemoprophylaxis and eight whose treatment did.ResultsThe mean direct cost of malaria treatment for patients without chemoprophylaxis was 1,776.0 EUR, and the mean indirect cost 524.2 EUR. In patients with chemoprophylaxis the mean direct cost was 405.6 EUR, and the mean indirect cost 257.4 EUR.ConclusionsThe analysis confirmed statistically-significant differences between the direct and indirect costs of treatment with and without chemoprophylaxis for patients with imported malaria.

Experimental Model of Allergic Asthma

The aim of the study was to prepare and evaluate the experimental model of allergic asthma. Changes in chough reflex, bronchoconstriction and the degree of inflammation were studied in ovalbumin (OVA) sensitized guinea pigs after 0, 7, 14, 21 days of exposure. The cough reflex was induced by citric acid inhalation in conscious animals in a double chamber body plethysmograph. Tracheal smooth muscle reactivity was assessed by examining the in vitro response to histamine (H) (10(-8)-10(-3) mol/l) and in vivo to H nebulization (10(-6) mol/l). BALF levels of IL-4, IL-5 and the eosinophil count were used as parameters of airway inflammation. After 7 days of OVA sensitization, there was an increase in tracheal smooth muscle contractility in vitro to cumulative concentration of H and an increase in cough parameters. After 14 days of OVA sensitization, there was a further increase in tracheal smooth muscle contractility to H, an increase in airway resistance, and a small increase in cough parameters. After 21 day of OVA sensitization, cough parameters were significantly reduced, airway resistance after H inhalation was increased, and there were significant increases in IL-4, IL-5, and eosinophils in BALF. In conclusion, progress in asthmatic inflammation during 21-day OVA sensitization caused a gradual increase in inflammatory mediators, a decline in cough reflex, and enhanced bronchoconstriction. This experimental model of allergic asthma can be used for pharmacological modulations of defense reflexes and inflammation.

Polyphenols and Their Components in Experimental Allergic Asthma

The aim of the study was to investigate the potential anti-inflammatory effects in -experimental allergic asthma of natural polyphenolic compounds or their single major components. The experiment was performed after 21-days sensitization of guinea pigs with ovalbumin suspension. Changes in airway reactivity after the long-term treatment with the polyphenolic compounds Provinol and Flavin-7 and their single major components quercetin and resveratrol during were assessed using a whole body plethysmography. Reactivity of tracheal smooth muscle was studied in vitro in response to cumulative doses of the bronchoconstrictive mediators histamine and acetylcholine. Furthermore, concentrations of the inflammatory cytokines IL-4 and IL-5 were measured in bronchoalveolar lavage fluid. The results demonstrate significant anti-inflammatory effects of Provinol and Flavin-7 exerted in the airways. In contrast, chronic treatment with quercetin and resveratrol, single components of the two polyphenols, did not show such activity. We conclude that polyphenolic compounds are more effective in the anti-inflammatory effects in the airways than their separate components.

Incidence of Haemophilus influenzae Type b Meningitis Among Children Less Than 5 Years of Age in Slovakia

Abstract Owing partly to a lack of disease burden data, Eastern and Central European countries have not introduced universal infant immunization against Haemophilus influenzae type b. To determine the incidence of Haemophilus influenzae type b meningitis among children less than 5 years of age, data were examined from eight districts in Slovakia that formed part of a national Haemophilus influenzae type b surveillance system. All invasive isolates of Haemophilus influenzae type b identified from these districts during 24 months (1996–1997) were sent to a single central laboratory for serotype confirmation. Thirty-five cases of confirmed Haemophilus influenzae type b meningitis were identified, for a disease incidence among children under 5 years of 17.3 per 100 000 population per year. Only 13 cases were identified during 1996, the first full year of the surveillance system. Records were available for review at six of the eight district laboratories and showed that, using World Health Organization definitions, almost half of the cases of probable bacterial meningitis were culture negative, suggesting that the true incidence of Haemophilus influenzae type b meningitis may be considerably higher than 17.3. The rate of Haemophilus influenzae type b meningitis among children less than 5 years of age in Slovakia is comparable to that found in Western Europe and North America during the pre-vaccine era. Thus, universal, publicly funded infant vaccination in Slovakia can be expected to have the same dramatic effect on Haemophilus influenzae type b disease morbidity and mortality as has been demonstrated in other countries that have adopted this approach.

Effects of Flavin7 on allergen induced hyperreactivity of airways

Some studies have suggested that the polyphenolic compounds might reduce the occurrence of asthma symptoms. The aim of our experiments was to evaluate the effects of 21 days of the flavonoid Flavin7 administration on experimentally induced airway inflammation in ovalbumin-sensitized guinea pigs. We assessed tracheal smooth muscle reactivity by an in vitro muscle-strip method; changes in airway resistance by an in vivo plethysmographic method; histological picture of tracheal tissue; and the levels of interleukin 4 (IL-4), and interleukin 5 (IL-5) in bronchoalveolar lavage fluid (BALF). Histological investigation of tracheal tissue and the concentrations of the inflammatory cytokines IL-4 and IL-5 in BALF were used as indices of airway inflammation. Administration of Flavin7 caused a significant decrease of specific airway resistance after histamine nebulization and a decline in tracheal smooth muscle contraction amplitude in response to bronchoconstricting mediators. Flavin7 minimized the degree of inflammation estimated on the basis of eosinophil calculation and IL-4 and IL-5 concentrations. In conclusion, administration of Flavin7 showed bronchodilating and anti-inflammatory effects on allergen-induced airway inflammation.

Human Dirofilariosis: The report of subcutaneous Dirofilaria repens infection in the Slovak Republic

SummaryDirofilariosis is a vector-borne disease that is spreading in Europe from the southern endemic regions to the northern countries, including Slovakia. The dog parasites Dirofilaria immitis and D. repens are zoonotic agents, responsible for the development of human pulmonary and subcutaneous dirofilariosis, respectively. The present paper reports the third case of human dirofilariosis in Slovakia caused by D. repens. The pacient, a 41-year-old woman, was referred with tumour process in the subcutaneous area of the right forearm. Within 14 days the USG confirmed the rapid increase of the nodule from 20 × 10 mm to 30 × 25 mm. The surgical extirpation of the tumour was indicated. Histological examination revealed the formation with eosinofilic rime and the presence of a worm in the centre, detected as D. repens.

Importance of complex diagnostic approach in differential diagnosis of alveolar echinococcosis

Summary Alveolar and cystic echinococcosis, serious parasitic diseases caused by larval stages of Echinococcus multilocularis and E. granulosus has been diagnosed in Slovakia for a long time. Study presents case of 49-years old patient with accidentally diagnosed one big (60 mm) and multiple small (2 - 24 mm) hypoechogenic structures localised in right liver lobe. According to positive serology to E. granulosus antigen and results of imaging examinations the patient was classifi ed as possible case of cystic echinococcosis and treated with mebendazole. Later, due to the worsening of clinical, laboratory and CT fi ndings surgical biopsy was performed and surgical biopsy and subsequent PCR examination of liver tissue confi rmed the diagnosis of alveolar echinococosis. Clinical picture of disease imitating cystic echinococcosis in presented case report, together with results of serological tests confi rmed importance of accurate differential diagnosis of echinococcosis. Each aspect of clinical and laboratory results should be considered responsibly, however, sometimes only molecular techniques can solve the problem of differential diagnosis.

Identification of Mycobacterium Species by MALDI-TOF Mass Spectrometry

Matrix-assisted laser desorption ionization - time-of-flight (MALDI-TOF) mass spectrometry enables to identify microorganisms by comparison of the protein content with reference spectra in the database. The aim of this study was to evaluate the efficacy of phenotypic identification of mycobacteria by MALDI-TOF mass spectrometry in laboratory practice. Seventy five isolates of mycobacteria were identified by molecular and phenotypic method, and the results were compared by MALDI-TOF. For MALDI-TOF, material was processed according to the Bruker Daltonics protocol and Mycobacterial Library database version 2.0, with 313 reference mycobacteria spectra. All except one of the 72 isolates agreed with regard to the species and genus by both methods. Forty three isolates were identified as the M. tuberculosis complex by MALDI-TOF. Thirty one isolates of nontuberculous mycobacteria were consistently identified by both methods to the species level. We conclude that MALDI-TOF mass spectrometry is an accurate method of bacterial identification. Simplicity, speed, and economic availability of the method makes it suitable for mycobacteria identification in a routine laboratory.

Two Distinct Patterns of Clostridium Difficile Diversity Across Europe Indicates Contrasting Routes of Spread.

Whole-genome sequencing of Clostridium difficile across Europe demonstrates 2 distinct transmission patterns. Healthcare-associated ribotypes showed genetic clustering by country and region, and widespread fluoroquinolone resistance. However, multiple common ribotypes showed no within-country clustering, consistent with Europe-wide dissemination via nonhealthcare routes/sources.