Kliničke i molekularno-epidemiološke karakteristike infekcije humanim metapneumovirusom u Hrvatskoj

Human metapneumovirus (HMPV) is an important respiratory pathogen, especially among young children. The aim of this study was to determine the incidence of HMPV infection in hospitalized children with acute respiratory tract infection (ARTI) in the season 2005/2006 in Croatia, as well as to perform the genotypic analysis of detected HMPV strains. From December 1 to March 31 2005/2006 nasopharyngeal secretions (NPSs) were collected from 402 inpatients up to 5 years of age with ARTI hospitalized in two clinics in Zagreb. NPSs were tested by real time RT-PCR assay targeting the nucleoprotein (N) gene of HMPV. To perform the phylogenetic study, partial nucleotide sequences were obtained for HMPV fusion (F) gene of 30 HMPV positive samples. HMPV was detected in 33 (8.2%), respiratory syncitial virus (RSV) in 101 (25.1%), adenoviruses in 41 (10.2%), parainfluenza viruses types1-3 in 24 patients (6.0%) and influenza viruses in 3 (0.7%) patients. The peak incidence of HMPV infection was detected in January (18/33). Three of 34 HMPV positive specimens were positive for other viral pathogens (1- parainfluenza virus type 3, 1- RSV, 1- adenovirus). No difference was noted between HMPV infected children and children infected with other viruses regarding to the sex. Children infected with adenovirus were significantly older than children infected with other viruses (HMPV, RSV and parainfluenza viruses). The majority of the infections caused by HMPV (69.7%) children presented with lower respiratory tract infections (LRTI). Comparative analysis of the clinical presentation of HMPV and RSV infected children revealed no significant differences between those two groups while adenoviruses and parainfluenza viruses caused more upper respiratory tract infections (URTI) than HMPV and RSV. Patient co-infected with parainfluenza virus type 3 and HMPV showed more severe clinical picture comparing to the clinical presentation of single-infection of HMPV in the same patient. Phylogenetic analysis showed the circulation of two main genetic lineages (A and B). It also showed the existence of two sublineages within the group B (B1 and B2) and three subclusters within lineage A (A1, A2a and A2b). The most prevalent subtype was the genotype B2 (15/30), followed by B1 (11/30), while group A of HMPV was detected in only four specimens. Interestingly, for one A2b genotype isolate epidemiological data suggests importation from Germany. All patients infected with HMPV genotype A had clinical presentation of LRTI, while 36% (9/25) of those infected with genotype B had URTI. All strains detected from patients infected with HMPV alone who required supplemental oxygen belonged to the genotype B2, while two HMPV strains from patients with co-infection who received supplemental oxygen belonged to the genotype B1. Evaluation of HMPV incidence contributes to the etiology of ARI in Croatia, while genotyping of the HMPV strains from Croatia contributes to the data of the genetic variability and molecular epidemiology of the virus in the region and in the world

Sociodemographic, sexual behaviour and microbiological profiles of men attending public health laboratory for testing for sexually transmitted diseases

In order to identify the groups at risk of sexually transmitted diseases (STDs), we assessed the sociodemographic profiles of men testing for STD, their sexual habits, and the results of microbiological analysis. During a three-year period, a total of 700 men older than 18 years of age completed the questionnaire regarding sociodemographic and sexual behavior. Urethral swabs were taken for microbiological analysis. Thirty-three percent of respondents reported not using condoms. Those that do not use condoms were predominantly less educated, unmarried but in steady relationships, employed, with children, and smokers. Alcohol or drug usage before sexual intercourse was disclosed by 21.4% of respondents, and 10.3% respondents reported sexual intercourses with commercial sex workers. Finally, 24.0% respondents reported sexual relations abroad. In 28.1% of subjects, one or more pathogens were observed in urethral swabs. The most commonly diagnosed microorganism was Ureaplasma urealyticum, followed by Chlamydia trachomatis, Mycoplasma hominis, Trichomonas vaginalis, and Neisseria gonorrhoeae. This study identified several factors that may contribute to the general risk of STD transmission, which will serve to better understand the transmission dynamics and implementation of adequate prevention programs. </p

Insights on the Antimicrobial Efficacy of Azithromycin According to the First Croatian Antibiotic Susceptibility Study of Chlamydia Trachomatis Isolates from Genitourinary Tract

Bez obzira na to što je rezistencija bakterija na antibiotike (antimikrobna otpornost) jedan od glavnih izazova moderne medicine, klamidijska je infekcija dosad pokazivala dobar klinički odgovor na lijek prvog izbora – azitromicin. Ipak, u literaturi postoje ponovljeni opisi laboratorijski dokazane rezistencije spolno prenosive bakterije Chlamydia trachomatis (C. trachomatis) koja uzrokuje ovu infekciju. Osim toga, Hrvatska je zemlja s visokom stopom propisivanja azitromicina i drugih antimikrobnih lijekova, što može utjecati na pojavu otpornosti ili slabijeg odgovora na terapiju. U skladu s time cilj prvoga hrvatskog istraživanja antimikrobnog profila C. trachomatis bio je utvrditi djelotvornost azitromicina i drugih antibiotika prema urogenitalnim kliničkim izolatima C. trachomatis u laboratorijskim uvjetima. U staničnome modelu McCoyevih stanica određivane su minimalna inhibitorna koncentracija (MIC) i minimalna klamidicidna koncentracija (MCC), a svi ispitivani sojevi bili su osjetljivi na azitromicin (MIC < 4 μg/ml). Premda rezistentni sojevi bakterije nisu nađeni, kod određenih izolata uočene su znatno više vrijednosti MCC-a za azitromicin, što pokazuje trend moguće smanjene osjetljivosti. Bez obzira na zahtjevnu metodologiju, potrebno je nastaviti s ovakvim istraživanjima, poglavito u zemljama s velikim opterećenjem propisivanja antibiotika.Regardless of the fact that antimicrobial resistance represents one of the most pressing issues of modern medicine, thus far chlamydial infection responded well to the first-line therapy i.e. azithromycin. Nonetheless, laboratory-confirmed resistance of sexually-transmitted pathogen Chlamydia trachomatis (C. trachomatis) causing this infection has been repeatedly described in the literature; furthermore, Croatia belongs to the group of countries that exhibit high prescribing rates of azithromycin and other antimicrobial drugs, which can, in turn, give rise to frank resistance or decreased susceptibility. Accordingly, the first Croatian antibiotic susceptibility study of Chlamydia trachomatis aimed to assess in vitro effectiveness of azithromycin and other antibiotics against clinical chlamydial isolates from genitourinary tract. McCoy cell culture model was used to assess minimal inhibitory concentration (MIC) and minimal chlamydicidal concentration (MCC), and all the tested isolates were susceptible to azithromycin (MIC < 4 μg/ml). Although no resistant strains have been found, certain isolates showed significantly higher values of MCC for azithromycin, pointing towards the trend of possible decreased susceptibility. Regardless of the complex methodology, it is pivotal to continue with this kind of resistance surveillance, especially in countries with high antibiotic prescribing rate

Molecular mechanisms of Chlamydia trachomatis resistance to antimicrobial drugs

Chlamydia trachomatis (C. trachomatis) is a leading cause of bacterial sexually transmitted infections in developed and undeveloped countries, and therefore a global public health issue. In an era of increasing bacterial resistance to antibiotics, resistance has been an exceedingly rare phenomenon in C. trachomatis; however, clinical treatment failures attributed to multidrug-resistant C. trachomatis strains have been described on several occasions. Cell culture systems using McCoy cells and subsequent immunofluorescent staining are still the most common methodology used for antimicrobial susceptibility testing, but the presence of resistance markers should be appraised by further genetic analysis. Azithromycin resistance of C. trachomatis is often a result of the mutations in the peptidyl transferase region of 23S rRNA genes, tetracycline resistance is usually linked to the presence of foreign genomic islands integrated in chlamydial chromosome, whereas a predominant mechanism of fluoroquinolone resistance is a point mutation in the gyrA quinolone-resistance-determining region. A nucleotide substitution in rpoB gene is responsible for rifampin resistance, and different mechanisms have been involved in the development of resistance to aminoglycosides, lincomycin and sulphonamide/trimethoprim combinations

Insights on the Antimicrobial Efficacy of Azithromycin According to the First Croatian Antibiotic Susceptibility Study of Chlamydia Trachomatis Isolates from Genitourinary Tract

Bez obzira na to što je rezistencija bakterija na antibiotike (antimikrobna otpornost) jedan od glavnih izazova moderne medicine, klamidijska je infekcija dosad pokazivala dobar klinički odgovor na lijek prvog izbora – azitromicin. Ipak, u literaturi postoje ponovljeni opisi laboratorijski dokazane rezistencije spolno prenosive bakterije Chlamydia trachomatis (C. trachomatis) koja uzrokuje ovu infekciju. Osim toga, Hrvatska je zemlja s visokom stopom propisivanja azitromicina i drugih antimikrobnih lijekova, što može utjecati na pojavu otpornosti ili slabijeg odgovora na terapiju. U skladu s time cilj prvoga hrvatskog istraživanja antimikrobnog profila C. trachomatis bio je utvrditi djelotvornost azitromicina i drugih antibiotika prema urogenitalnim kliničkim izolatima C. trachomatis u laboratorijskim uvjetima. U staničnome modelu McCoyevih stanica određivane su minimalna inhibitorna koncentracija (MIC) i minimalna klamidicidna koncentracija (MCC), a svi ispitivani sojevi bili su osjetljivi na azitromicin (MIC < 4 μg/ml). Premda rezistentni sojevi bakterije nisu nađeni, kod određenih izolata uočene su znatno više vrijednosti MCC-a za azitromicin, što pokazuje trend moguće smanjene osjetljivosti. Bez obzira na zahtjevnu metodologiju, potrebno je nastaviti s ovakvim istraživanjima, poglavito u zemljama s velikim opterećenjem propisivanja antibiotika.Regardless of the fact that antimicrobial resistance represents one of the most pressing issues of modern medicine, thus far chlamydial infection responded well to the first-line therapy i.e. azithromycin. Nonetheless, laboratory-confirmed resistance of sexually-transmitted pathogen Chlamydia trachomatis (C. trachomatis) causing this infection has been repeatedly described in the literature; furthermore, Croatia belongs to the group of countries that exhibit high prescribing rates of azithromycin and other antimicrobial drugs, which can, in turn, give rise to frank resistance or decreased susceptibility. Accordingly, the first Croatian antibiotic susceptibility study of Chlamydia trachomatis aimed to assess in vitro effectiveness of azithromycin and other antibiotics against clinical chlamydial isolates from genitourinary tract. McCoy cell culture model was used to assess minimal inhibitory concentration (MIC) and minimal chlamydicidal concentration (MCC), and all the tested isolates were susceptible to azithromycin (MIC < 4 μg/ml). Although no resistant strains have been found, certain isolates showed significantly higher values of MCC for azithromycin, pointing towards the trend of possible decreased susceptibility. Regardless of the complex methodology, it is pivotal to continue with this kind of resistance surveillance, especially in countries with high antibiotic prescribing rate

Značaj IgG aviditeta u dijagnostici infekcije citomegalovirusom u trudnica i novorođenčadi

Aim: To asses the value of IgG avidity in diagnosis of CMV infection in pregnant women and newborn infants. Methods: During the three-year period (2003-2005) serum samples from 64 pregnant women and 32 infants less than 12 months of age with suspected congenital/perinatal CMV infection were collected. Sera were tested for CMV IgM and IgG using an indirect enzyme-linked immunosorbent assay and IgG avidity using urea as denaturing agent. Results: Among IgM positive women, 2/26 (7.6%) showed an increase of IgG avidity index (AI) from intermediate to high AI in paired sera samples indicating recent primary infection and 24/26 (92.4%) showed high AI indicating past infection. All women with negative IgM antibodies had high AI. In infants less than 12 months old, acute/recent primary CMV infection was documented in 8/12 (66.7%) children with positive IgM and in 10/20 (50.0%) children with negative IgM antibodies. In two children less than three months with high AI, CMV infection was confirmed by virus isolation from urine. One of them showed a decrease of AI from high to low in paired sera samples. Conclusions: IgG avidity differentiates primary from nonprimary CMV infection in both pregnant women and infants older than three months. In children less than 3 months of age, transplacentally transferred maternal antibodies of high avidity may influence on the serologic test results.Cilj rada: Ispitati vrijednost IgG aviditeta u dijagnostici CMV infekcije u trudnica i novorodencadi. Metode: Tijekom trogodišnjeg perioda (2003-2005) prikupljeni su uzorci seruma od 64 trudnice, te 32 djece do godinu dana starosti sa suspektnom kongenitalnom/perinatalnom CMV infekcijom. Serumi su testirani na prisustvo CMV IgM i IgG protutijela metodom ELISA, te aviditet IgG protutijela uz primjenu uree. Rezultati: U skupini trudnica s pozitivnim IgM protutijelima, u njih 2/26 (7,6%) dokazan je porast indeksa IgG aviditeta (AI), od granicnog u visoki AI u parnim uzorcima seruma, što ukazuje na nedavnu primarnu CMV infekciju, dok je 24/26 (92,4%) imalo visok AI (prošla CMV infekcija). Sve trudnice s negativnim IgM protutijelima imale su visok AI. U djece mlade od 12 mjeseci, akutna/nedavna primarna CMV infekcija dokazana je u 8/12 (66,7%) djece s pozitivnim IgM, te u 10/20 (50,0%) djece s negativnim IgM protutijelima. U dvoje djece mlade od 3 mjeseca s visokim AI, CMV infekcija je potvrdena izolacijom virusa iz urina. U jednog od njih došlo je do pada AI iz visokog u niski, u parnom uzorku seruma. Zakljucci: Pomocu testa IgG aviditeta moguce je razluciti primarnu od prošle CMV infekcije u trudnica i djece starije od 3 mjeseca. U djece mlade od 3 mjeseca, transplacentarno prenesena majcina IgG protutijela visokog aviditeta mogu utjecati na rezultate seroloških pretraga

Tehnički aspekti testiranja antimikrobne osjetljivosti bakterije Chlamydia trachomatis u staničnoj kulturi

Chlamydia trachomatis (C. trachomatis) is the most common bacterial agent of sexually transmitted infections and an obligate intracellular bacterium with a unique life cycle. Cell culture systems using McCoy cells and immunofluorescent staining to identify characteristic chlamydial inclusions represent the most common methodology employed in antimicrobial susceptibility testing of this prevalent pathogen. The testing process can be broken down into four technical steps – preisolation stage, isolation of C. trachomatis in cell culture, preparatory steps for antimicrobial susceptibility testing and, finally, testing phase with a determination of minimal inhibitory concentration (MIC) and minimal chlamydicidal concentration (MCC). As this technique requires precision and careful attention to conditions which may influence both the ability of chlamydial organisms to infect cells in culture and the efficacy of a tested antibiotic through mechanisms such as intracellular uptake, following these steps is pivotal in order to ensure reproducibility and comparability of obtained results.Chlamydia trachomatis (C. trachomatis) najčešći je bakterijski uzročnik spolno-prenosivih infekcija i obligatna unutarstanična bakterija s jedinstvenim životnim ciklusom. Za testiranje antimikrobne osjetljivosti ovog učestalog patogena danas se najčešće koristi stanična kultura McCoy stanica uz imunofluorescentno bojanje za detekciju karakterističnih klamidijskih inkluzija. Sam postupak testiranja može se podijeliti u četiri osnovna koraka – predizolacijska faza, izolacija C. trachomatis u staničnoj kulturi, priprema za testiranje antimikrobne osjetljivosti te, najzad, faza testiranja uz određivanje minimalne inhibitorne koncentracije (MIC) i minimalne klamicidne koncentracije (MCC). Kako ova metoda zahtjeva preciznost i posvećivanje posebne pozornosti čimbenicima koji mogu utjecati na sposobnost klamidija da inficiraju stanice u kulturi, ali i na učinkovitost testiranog antibiotika uslijed različitih mehanizama kao što je pravodobni ulazak u stanicu, pravilno izvođenje svakog od ovih koraka je ključno kako bi se osigurala reproducibilnost i usporedivost dobivenih rezultata

DIE HAUFIGKEIT POSTTRAUMATISCHER STRESSREAKTIONEN UNTER VERTRIEBENEN UND DER MOGLICHE BEZUG ZUR INHAFTIERUNG IN GEFANGENENLAGERN

Cilj rada bio je utvrditi učestalost PTSP-a u prognanika iz Hrvatske te ispitati povezanost PTSP-a s brojem trauma, posebno s boravkom u logoru, te s demografskim varijablama. Ispitana je grupa od 88 prognanika, od kojih 34 muškarca i 54 žene, prosječne dobi 43 godine. Primijenjeni su upitnik za traume i Watsonov intervju za PTSP. Grupa je proživjela prosječno tri traume, a učestalost PTSP-a nakon dvije godine života u progonstvu iznosila je 45,45%. Ustanovljeno je da se pojavnost ukupne izraženosti simptomatike PTSP-a povećava s povećanjem broja traumatskih događaja i s dobi osobe, a smanjuje se s većim obrazovanjem osobe. Grupa prognanika (N=18) koji su bili zatočeni u logoru do najviše mjesec dana, a koji nisu bili izloženi torturi, uspoređena je s komparativnom grupom prognanika (N=18) izjednačenom po dobi, spolu, obrazovanju i broju proživljenih trauma. Grupe se nisu statistički značajno razlikovale u izraženosti ukupne simptomatike PTSP-a. Zaključno, rezultati ukazuju da su mlađa odrasla dob i obrazovanje protektivni čimbenici u reakciji na traumu. Kratko zatočeništvo u logoru, bez torture, nije imalo zaseban efekt na pojavnost simptomatike PTSP-a u politraumatiziranoj grupi prognanika.The aim of the study was to explore the prevalence of PTSD in refugees from Croatia and investigate the relationship between PTSD and the number of experienced traumas, especially with regard to being held in detention camp, as well as the relationship between PTSD and demographic variables. A group of refugees, consisting of 34 men and 54 women, mean age 43, were studied. Trauma Questionnaire and Watson\u27s PTSD Interview were applied. The group experienced three traumas, on average; whereas after two years of life in captivity the prevalence of PTSD was 45.45%. It was established that the total of PTSD symptoms grow with the increasing number of traumas and with the refugee\u27s age, but decrease with the person\u27s higher education. The group of refugees (N=18) who were kept in detention camp no longer than one month and were not tortured, was compared with the comparative refugee group (N=18), equal in gender, age, education and number of traumas experienced. The groups had shown statistically the same amount of total PTSD symptomatics. Conclusively, the results indicate that young adulthood and higher education are protective factors in reaction to trauma. A short period of time spent in detention camp, without torture, did not have a separate effect on PTSD symptomatics in the group of po- Iytraumatized adult refugees.Die vorliegende Arbeit soll ermitteln, wie haufig posttraumatische Stressreaktionen unter kroatischen Vertriebenen auftreten und inwiefern diese Stressreaktionen mit der Zahl der durch- Iebten Traumata, insbesondere mit der Inhaftierung in Gefangenenlagern sowie mit demographischen Variablen in Verbindung stehen. Die befragte Personengruppe umfasste 88 Vertriebene: 34 Manner und 54 Frauen im Durchschnittsalter von 43 Jahren. Angewandt wurden ein Fragebagen zur Ermittlung erlebter Traumata sowie der Watsonsche Fragebagen zur Ermittlung posttra umatischer Stressreaktionen. Die befragten Personen erlebten im Durchschnitt jeweils 3 Traumata, die Haufigkeit posttraumatischer Stressreaktionen zwei Jahre nach der Vertreibung betrug 45,45%. Es wurde festgestelIt, dass sich mit der Zahl traumatischer Erlebnisse und mit dem wachsenden Alter der Befragten die Gesamtauspraqunq des posttraumatischen Stresssyndroms steigert, mit einem htiheren Bildungsgrad jedoch abfallt, Eine Gruppe von Vertriebenen (N=18), die bis zu 1 Manat in Lagern inhaftiert gewesen, jeda ch keinen Folterungen ausgesetzt waren, wurde mit einer Vergleichsgruppe von Vertriebenen (N=18) kompariert, mit der sie in Alter, Geschlecht, Bildungsgrad und der Zahl durchlebter Traumata Obereinstimmte. Die Gruppen unterschieden sich statistisch nur wenig in der Gesamtauspragung posttraumatischer Stressreaktionen. Es kann gemar.. den Resuitaten abschliefšend gesagt werden, dass ein jOngeres Erwachsenenalter und ein h6herer Bildungsgrad Schutzfaktoren bei der Reaktion auf Traumata darstellen. Kurze Lageraufenthalte ohne Folterungen hatten keinerlei Sonderwirkung auf die Haufigkeit posttraumatischer Stressreaktionen unter den befragten polytraumatisierten Vertriebenen