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U radu opisujemo mladu bolesnicu sa šećernom bolesti tipa I koja se zarazila virusom hepatitisa A (HAV) genotipa IB tijekom boravka u Burkini Faso. Prikaz bolesnice nedvojbeno potvrđuje važnost pridržavanja preporuka stručnih društava i nadležnih tijela o cijepljenju putnika., We describe a clinical case of hepatitis A in a young women with type I diabetes infected with genotype IB of hepatitis A virus (HAV) from Burkina Faso. The case confirms the importance of strict adherence to the recommendations by professional associations and institutions regarding vaccination of the travellers.

Epstein-Barr virus (EBV) is a widely disseminated herpesvirus for which antibodies have been demonstrated in over 90% of adults worldwide. After subclinical primary EBV infections, as well as after infectious mononucleosis, the virus can be shed in saliva for a prolonged period of time. Diseases and disorders that can induce EBV salivary shedding include mental disorders and sex, connective tissue disease, multiple sclerosis, systemic lupus erythematosus, malaria and HIV infection. As the occurrence of EBV in saliva during acute infectious diseases has not been systematically researched so far, this pilot cross-sectional study aimed to investigate the possible relationship between acute infectious diseases and salivary shedding of EBV. A total of 40 patients with acute infectious diseases was enrolled, along with 41 adults free of acute infections. Peripheral venous blood samples for serodiagnosis and saliva samples for EBV PCR testing were collected from both groups. The most common acute infectious disease was COVID-19 pneumonia, followed by haemorrhagic fever with renal syndrome. Crude proportions of people with positive serological test results and those with saliva viral shedding were similar in the two groups. The presented preliminary data does not indicate acute infectious conditions as a marked “contributor” in increasing salivary EBV shedding.

Hepatitis C vodeći je uzrok kronične bolesti jetre, hepatocelularnog karcinoma i važna indikacija za transplantaciju jetre. Glavni cilj liječenja je postizanje trajnog virološkog odgovora koji označava negativnu viremiju 12 tjedana nakon završetka liječenja (SVR, od engl. sustained virological response). Uvođenjem antivirusnih lijekova s izravnim djelovanjem, povećala se stopa SVR-a, skratilo vrijeme trajanja terapije i utjecalo se na stadij fibroze i steatoze jetre nakon liječenja. Cilj ovog rada bio je analizirati promjenu stadija fibroze (F1-F4) i steatoze (S0-S3) primjenom tranzijentne elastografije u prethodno neliječenih bolesnika s kroničnim hepatitisom C (n=205) koji su postigli trajni virološki odgovor, a bili liječeni isključivo direktno djelujućim antivirusnim lijekovima (DAA).

During the first half of 2022, the World Health Organization reported an outbreak of acute severe hepatitis of unknown aetiology (AS-Hep-UA) in children, following initial alerts from the United Kingdom (UK) where a cluster of cases was first observed in previously well children aged

RNA viruses are characterised by extremely high genetic variability due to fast replication, large population size, low fidelity, and (usually) a lack of proofreading mechanisms of RNA polymerases leading to high mutation rates. Furthermore, viral recombination and reassortment may act as a significant evolutionary force among viruses contributing to greater genetic diversity than obtainable by mutation alone. The above-mentioned properties allow for the rapid evolution of RNA viruses, which may result in difficulties in viral eradication, changes in virulence and pathogenicity, and lead to events such as cross-species transmissions, which are matters of great interest in the light of current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemics. In this review, we aim to explore the molecular mechanisms of the variability of viral RNA genomes, emphasising the evolutionary trajectory of SARS-CoV-2 and its variants. Furthermore, the causes and consequences of coronavirus variation are explored, along with theories on the origin of human coronaviruses and features of emergent RNA viruses in general. Finally, we summarise the current knowledge on the circulating variants of concern and highlight the many unknowns regarding SARS-CoV-2 pathogenesis.

West Nile Virus Neuroinvasive Disease (WNV NID) requires prolonged intensive care treatment, resulting in high mortality and early disability. Long-term results are lacking. We have conducted an observational retrospective study with a prospective follow-up of WNV NID patients treated at the Intensive Care Unit (ICU), University Hospital for Infectious Diseases, Zagreb, Croatia, 2013–2018. Short-term outcomes were vital status, length of stay (LOS), modified Rankin Scale (mRS), and disposition at discharge. Long-term outcomes were vital status and mRS at follow-up. Twenty-three patients were identified, 78.3% males, median age 72 (range 33–84) years. Two patients (8.7%) died in the ICU, with no lethal outcomes after ICU discharge. The median ICU LOS was 19 days (range 5–73), and the median hospital LOS was 34 days (range 7–97). At discharge, 15 (65.2%) patients had moderate to severe/mRS 3–5, 6 (26.0%) had slight disability/mRS 2–1, no patients were symptom-free/mRS 0. Ten (47.6%) survivors were discharged to rehabilitation facilities. The median time to follow-up was nine months (range 6–69). At follow-up, seven patients died (30.5%), five (21.7%) had moderate to severe/mRS 3–5, one (4.3%) had slight disability/mRS 2–1, six (26.1%) had no symptoms/mRS 0, and four (17.4%) were lost to follow-up. Briefly, ten (43.5%) survivors improved their functional status, one (4.3%) was unaltered, and one (4.3%) aggravated. In patients with severe WNV NID, intensive treatment in the acute phase followed by inpatient rehabilitation resulted in significant recovery of functional status after several months.

The hepatitis A virus (HAV) is a highly hepatotropic virus transmitted mainly via the fecal–oral route. The purpose of this study is to describe a prolonged HAV outbreak in HIV-infected men who have sex with men (MSM) and pre-exposure prophylaxis (PrEP) users in Croatia in 2022. Croatia has a centralized system of HIV care and the PrEP service is only available at the University Hospital for Infectious Diseases (UHID), Zagreb. We reviewed all MSM living with HIV and MSM PrEP users at UHID and identified those diagnosed with HAV between January and October 2022. During this period, a total of 1036 MSM living with HIV and 361 PrEP users were followed, and 45 (4.4%) and 32 (8.9%) were diagnosed with HAV, respectively. Most cases were diagnosed in mid-February. A total of 70.1% (726/1036) MSM living with HIV and 82.3% (297/361) PrEP users were susceptible to HAV. Sequencing information was available for 34 persons; in all cases the HAV subtype IA was found. Our findings indicate that both MSM living with HIV and HIV-uninfected PrEP users are vulnerable to HAV infection and might be a potential source for a more widespread HAV epidemic.

Identifying individuals recently infected with HIV has been of great significance for close monitoring of HIV epidemic dynamics. Low HIV sequence ambiguity (SA) has been described as a promising marker of recent infection in previous studies. This study explores the utility of SA defined as a proportion of ambiguous nucleotides detected in baseline pol sequences as a tool for routine real-time surveillance of HIV incidence trends at a national level. A total of 353 partial HIV-1 pol sequences obtained from persons diagnosed with HIV infection in Slovenia from 2000 to 2012 were studied, and SA was reported as a percentage of ambiguous base calls. Patients were characterized as recently infected by examining anti-HIV serological patterns and/or using commercial HIV-1 incidence assays (BED and/or LAg-Avidity assay). A mean SA of 0.29%, 0.14%, and 0.19% was observed for infections classified as recent by BED, LAg, or anti-HIV serological results, respectively. Welch's t-test showed a significant difference in the SA of recent versus long-standing infections (p < 0.001). CD4+ T-cell counts ≤250 cells/mm3 significantly correlated with higher SA (p < 0.001), whereas the homo/bisexual transmission route significantly correlated with lower SA (p = 0.005). When the LAg-assay was used as an indicator of recent infection, a receiver operating characteristic curve with the largest area under the curve (0.896) was observed for SA (sensitivity and specificity of 79%), indicating the best correlation of the data. A reliable estimation of the trends of HIV incident infection could be inferred from measuring SA irrespective of the cutoff used ; however, in Slovenia it seems that lower cutoffs are more appropriate. Our data suggest that SA could be used as a real-time surveillance tool for close monitoring of HIV incidence trends, especially in countries where baseline HIV resistance genotyping is performed routinely, rendering this approach cost-effective.

Epstein-Barr virus (EBV) or human herpesvirus 4 (HHV-4) is a ubiquitous human oncogenic virus, and the first human virus found to express microRNAs (miRNAs). Its genome contains two regions encoding more than 40 miRNAs that regulate expression of both viral and human genes. There are numerous evidences that EBV miRNAs impact immune response, affect antigen presentation and recognition, change T- and B-cell communication, drive antibody production during infection, and have a role in cell apoptosis. Moreover, the ability of EBV to induce B-cell transformation and take part in mechanisms of oncogenesis in humans is well known. Although EBV infection is associated with development of various diseases, the role of its miRNAs is still not understood. There is abundant data describing EBV miRNAs in nasopharyngeal carcinoma and several studies that have tried to evaluate their role in gastric carcinoma and lymphoma. This review aims to summarize so far known data about the role of EBV miRNAs in altered regulation of gene expression in human cells in EBV-associated diseases.

Objective: Hepatitis C virus (HCV) genotyping is an important part of pre-treatment diagnostic algorithms as it guides the choice of therapeutic regimens. The aim of this study was to analyse the distribution of HCV genotypes in patients with chronic hepatitis C from Croatia in the period 2008-2015. Methods: The study enrolled 3, 655 anti-HCV positive patients with available results of HCV genotyping from the three largest national HCV genotyping laboratories. Results: The majority of HCV-infected individuals enrolled in the study were male (70.7%). Analysis of age distribution in a subset of 2, 164 individuals showed a mean age of 40.9 years (SD 11.77 years). Croatian patients were mostly infected with HCV genotype 1 (56.6%), followed by genotype 3 (37.3%), genotype 4 (4.2%) and genotype 2 (1.8%). Genotype 1 subtyping in a subset of 1, 488 patients showed 54% (803/1, 488) of 1b infections and 46% (685/1, 488) of 1a infections. Percentages of genotype 1 were the highest in Central/Northwestern and Eastern Croatia and the lowest in the Central/Southern Adriatic Region. Genotype 3 was most frequently found in the Central/Southern Adriatic Region (49.1%) but represented only 17.5% of infections in Eastern Croatia (p < 0.001). Conclusions: The results of this nine-year retrospective analysis on the distribution of HCV genotypes and subtypes in 3, 655 HCV-infected individuals from Croatia showed that the majority of infections can be attributed to genotypes 1 and 3 with absence of major changes in the molecular epidemiology of the two most frequent HCV genotypes infection in Croatia in the past 20 years.

Objectives: Kosovo’s current health care system does not support organized nationwide cervical cancer screening and human papillomavirus (HPV) vaccination programs. To date, no reliable data are available on cervical cancer incidence and mortality in Kosovo, or on high-risk HPV (HR-HPV) prevalence and HPV type distribution. Our aim is to determinate the pre-vaccination prevalence and distribution of HR-HPVs and to assesses the associations between sociodemographic characteristics and increased risk of HPV infection in women from Kosovo. Material and methods: Detection of HR-HPV DNA in cytologically evaluated cervical smears was performed using a clinically validated Abbott RealTime High Risk HPV test, Roche Linear Array HPV Genotyping Test, HPV52 type-specific real-time PCR and an in-house GP5+/GP6+/68 PCR. Results: The crude overall prevalence of any of the HR-HPVs was estimated at 13.1% (26/199; 95% confidence interval (CI): 9.1–18.5%), with HPV16 being the most common type (7/26, 26.9%), followed by HPV31 and HPV51, each detected in 4/26 (15.4%) cervical specimens, HPV18, detected in 3/26 (11.5%) specimens, HPV52 and HPV66, each detected in 2/26 (7.7%) specimens, and HPV33, HPV45, HPV56, and HPV58, each detected in a single (3.9%) specimen. Women over 40 (OR = 0.36), older than 18 at sexual debut (odds ratio (OR) = 0.28), those that had delivered at least one child (OR = 0.32), and those that had a history of pregnancy termination (OR = 0.39) were at lower risk for HPV infection. Conclusion: Because more than 70% of cervical precancerous lesions could have been prevented in Kosovo using nationwide HPV-based cervical cancer screening and HPV vaccination, it is of outmost importance to implement both programs in the national health care system as soon as possible.

We report an HIV-infected person who was treated for lymphogranuloma venereum cervical lymphadenopathy and proctitis in Croatia in 2014. Infection with a variant L2b genovar of Chlamydia trachomatis was detected in a cervical lymph node aspirate. A prolonged course of doxycycline was required to cure the infection.

Hepatitis C vodeći je uzrok kronične bolesti jetre, hepatocelularnog karcinoma i važna indikacija za transplantaciju jetre. Glavni cilj liječenja je postizanje trajnog virološkog odgovora koji označava negativnu viremiju 12 tjedana nakon završetka liječenja (SVR, od engl. sustained virological response). Uvođenjem antivirusnih lijekova s izravnim djelovanjem, povećala se stopa SVR-a, skratilo vrijeme trajanja terapije i utjecalo se na stadij fibroze i steatoze jetre nakon liječenja. Cilj ovog rada bio je analizirati promjenu stadija fibroze (F1-F4) i steatoze (S0-S3) primjenom tranzijentne elastografije u prethodno neliječenih bolesnika s kroničnim hepatitisom C (n=205) koji su postigli trajni virološki odgovor, a bili liječeni isključivo direktno djelujućim antivirusnim lijekovima (DAA)., Hepatitis C is the leading cause of chronic liver disease, hepatocellular carcinoma and an important indication for liver transplantation. The main objective of the treatment is to achieve a sustained virological response defined as a negative HCV RNA 12 weeks after completion of therapy (SVR). Direct-acting antivirals have been developed to treat chronic HCV infection. They have resulted in increased SVR rates, shorter and simpler treatment regimens, and minimal adverse effects. The aim of this study was to analyze the change in the stage of fibrosis (F1-F4) and steatosis (S0-S3) using transient elastography in treating naive patients with chronic hepatitis C (n = 205) who achieved a sustained virological response and were treated exclusively with directly acting antiviral drugs (DAA).

Molecular epidemiology of HIV-1 infection in treatment-naive HIV-1 infected persons from Croatia was investigated. We included 403 persons, representing 92.4% of all HIV-positive individuals entering clinical care in Croatia in 2014–2017. Overall prevalence of transmitted drug resistance (TDR) was estimated at 16.4%. Resistance to nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside RTI (NNRTIs) and protease inhibitors (PIs) was found in 11.4%, 6.7% and 2.5% of persons, respectively. Triple-class resistance was determined in 2.2% of individuals. In addition, a single case (1.0%) of resistance to integrase strand-transfer inhibitors (InSTIs) was found. Deep sequencing was performed on 48 randomly selected samples and detected additional TDR mutations in 6 cases. Phylogenetic inference showed that 347/403 sequences (86.1%) were part of transmission clusters and identified forward transmission of resistance in Croatia, even that of triple-class resistance. The largest TDR cluster of 53 persons with T215S was estimated to originate in the year 1992. Our data show a continuing need for pre-treatment HIV resistance testing in Croatia. Even though a low prevalence of resistance to InSTI was observed, surveillance of TDR to InSTI should be continued.

Resistance of Human Immunodeficiency Virus (HIV) to antiretroviral drugs is an important limitation in achieving complete suppression of viral replication and therefore represents an important clinical issue. It refers especially to therapy-naive individuals infected with resistant HIV strains, e.g. individuals with transmitted drug resistance (TDR). Transmitted drug resistance mutations (TDRMs) are clinically relevant and may reduce the efficacy of antiretroviral therapy. In this paper, we report the first case of HIV-1 transmitted triple-class drug resistance in Croatia. The aim of this study was to characterize drug resistance patterns and TDRMs in the newly diagnosed, treatment-naive HIV-1 patient with such a complex resistance pattern. Sanger sequencing (SS) of the sample showed four reverse transcriptase inhibitor (RTI) resistance mutations (E44D, T215E, K103N, L100I) affecting two drug classes and two protease inhibitor resistance mutations (V32I, I47V). To characterize HIV-1 minority drug resistance variants below the detection limit of SS, deep sequencing (DS) analysis was performed. DS analysis identified the same triple class resis- tance pattern that was identified by SS with addition of several other RTI mutations. The patient described in this report is the first patient with HIV-1 triple-class resistance in Croatia and further studies will be directed toward analysing possible local onward transmission of this resistant virus., Rezistencija virusa humane imunodeficijencije (HIV) na antiretrovirusne lijekove sprječava supresiju virusne replikacije te predstavlja značajan izazov u kliničkoj medicini. Posebno valja istaknuti problem primarne rezistencije (engl. transmitted drug resistance, TDR) koja se odnosi na prethodno neliječene osobe koje su zaražene rezistentnim sojevima HIV-a. Mutacije koje su pov- ezane s primarnom rezistencijom (engl. transmitted drug resistant mutations, TDRM) su klinički značajne i mogu nepovoljno djelovati na učinkovitost antiretrovirusnog liječenja. U ovom je radu opisana prva osoba s primarnom rezistencijom HIV-a na 3 klase antiretrovirusnih lijekova u Hr- vatskoj. Cilj ovog istraživanja bio je analizirati obrasce primarne rezistencije i TDRM u novodi- jagnosticiranog i neliječenog HIV-om zaraženog pojedinca. Primjenom Sangerovog sekvenciranja (SS) dokazali smo četiri mutacije povezane s rezistencijom na inhibitore reverzne transkriptaze (E44D, T215E, K103N, L100I) koje smanjuju osjetljivost na dvije klase lijekova (nukleozidne analoge inhibitore reverzne transkriptaze i nenukleozidne inhibitore reverzne transkriptaze) kao i dvije mutacije (V32I, I47V) povezane s rezistencijom na inhibitore proteaze. U svrhu identifikacije mogućeg postojanja manjinskih rezistentnih varijante ispod granice detekcije SS-a, provedena je analiza dubinskim sekvenciranjem (DS). DS analiza identificirala je isti obrazac rezistencije na 3 klase antiretrovirusnih lijekova identificiran s SS uz nekoliko dodatnih mutacija. U ovom je radu opisan prvi slučaj primarne rezistencije HIV-a na 3 klase antiretrovirusnih lijekova, a buduća istraživanja analizirat će moguće putove transmisije ovog rezistentnog virusa u Hrvatskoj.

A 70-year-old patient was admitted to the Department of Oral Medicine for multiple oral ulcerations on the left buccal mucosa, around 0.5 cm in diameter, as well as on the gingiva. Otherwise, the patient suffered from chronic lymphocytic leukemia, hypogammaglobulinemia, chronic renal insufficiency, with complete afunction of the right kidney, asthma, hypertension, gastritis and prostate hyperplasia. Differential diagnosis of oral ulcerations included drug induced oral ulcerations, paraneoplastic pemphigus, viral ulcerations (cytomegalovirus, herpes simplex viruses), fungal ulcerations (candidiasis, aspergillosis, histoplasmosis, cryptococcosis) and bacterial ulcerations, as well as neutropenic ulcers. One of the possible explanations was that the lesions were due to the use of drugs, the more so as oral lesions evolved when the doses of allopurinol and chlorambucil were increased, and subsided when the doses of both drugs were decreased. However, we could not establish for sure whether the lesions were due to allopurinol or chlorambucil. According to literature data, allopurinol is one of the most frequent drugs known to induce skin adverse reactions, therefore we assumed that it was the culprit drug. Unfortunately, several weeks later the patient died from sepsis, pneumonia with respiratory insufficiency and multiorgan failure.

Background and aims: Genetic polymorphisms of immune mediators have been associated with differences in the natural course of chronic hepatitis C (CHC). The aim of this study was to analyze the association of IL-1β gene polymorphism with the stage of liver fibrosis (LF), grade of necroinflammatory activity (NIA) and fibrosis progression rate (FPR) in CHC patients. Patients and methods: The study included 50 treatment-naive Croatian CHC patients (36 male and 14 female; age median 37.5 years) with elevated ALT. Diallele polymorphism (C/T) at locus -31 in the IL-1β gene promoter region was determined by restriction fragment length polymorphism (RFLP). Stage of LF and NIA were assessed from liver biopsy sample according to Ishak classification. Results: There was no difference in the stage of LF and NIA level between particular patient genotypes. However, patients with at least 1 C allele at locus -31showed significantly faster FPR than those with no C allele (0.4 vs. 0.258 Ishak's units/year; p = 0.043). Higher stages of fibrosis were observed in older patients (p = 0.001) and those infected at an older age (p = 0.017). Conclusion: Our study demonstrated that the carriage of at least 1 C allele at -31 locus of IL-1β gene led to faster progression of LF in CHC patients with a biochemically active disease, but did not determine the final stage of fibrosis development. Combined with other risk factors, this finding may serve as a genetic marker to identify patients that require earlier introduction of therapy, since delay could hamper therapeutic success due to rapid disease progression.

U razdoblju od 2008. do 2017. u Klinici za infektivne bolesti "Dr. Fran Mihaljević" u Zagrebu liječeno je 210 bolesnika s kroničnom HBV infekcijom koji su imali histološku i biokemijsku aktivnost bolesti. Stadij fibroze određivan je patohistološki i elastografski. Cirozu je već u početku imalo 14,5 % bolesnika. Iako je funkcionalno izlječenje bilo rijetko (5,5 % slučajeva) kod većine bolesnika postignuta je supresija virusne replikacije, usporavanje ili prekid razvoja fibroze, pa čak i regresija stadija fibroze za jedan stupanj po Metaviru u 16 % bolesnika. Zapaženo je smanjenje frekvencije pojave dekompenzirane ciroze i hepatocelularnog karcinoma za 2 do 3 puta u usporedbi s podacima kod neliječenih bolesnika. Bolesnici liječeni lamivudinom imali su visoku incidenciju razvoja rezistencije (32 % nakon 5 godina liječenja), te je terapija nastavljena tenofovirom uz većinom dobru podnošljivost i bez razvoja rezistencije. HBeAg nije se pokazao kao značajan prediktor težine i toka bolesti. Značajan je broj hematoloških i drugih imunosuprimiranih bolesnika (n = 24), koji terapiju primaju radi prevencije reaktivacije latentne HBV infekcije., A total of 210 patients with chronic HBV infection and histological and biochemical activity of disease were treated at the University Hospital for Infectious Diseases "Dr. Fran Mihaljević" in Zagreb in the period from 2008 to 2017. The fibrosis stage was determined histologically and by transient elastography, and cirrhosis was found initially in 14,5 % of the patients. Although functional recovery was rare (5.5 % of cases), the suppression of viral replication, slowing down or discontinuation of fibrosis progression was achieved in most of the treated. Even fibrosis regression by one stage in Metavir scale was achieved in 16 % of patients. There was also a decrease in the frequency of development of decompensated cirrhosis (2 patients) and hepatocellular carcinoma (2 patients) by 2 to 3 times in comparison to historical controls in nontreated patients. Patients treated with lamivudine had a high incidence of resistance development (32 % after 5 years). They continued therapy with tenofovir with good tolerability and no resistance development. HBeAg did not seem to be a significant predictor for the severity and the course of the disease. There is also a significant number of hematologic and other immunosuppressed patients (n = 24) receiving treatment to prevent the reactivation of latent HBV infection.

Slovenia is a small European country with a total of 547 HIV-infected individuals cumulatively reported by the end of 2011. However, the estimated incidence rate of HIV infections increased from 7.0 per million in 2003 to 26.8 per million in 2011. In this study, we assessed the prevalence of transmitted drug resistance (TDR) in the past 6 years (2005-2010) and analyzed the time trend of the proportion of men having sex with men (MSM) and HIV-1 subtype B among newly diagnosed individuals in a 15-year period (1996-2010) in Slovenia. Among 150 patients included in the study, representing 63% of HIV-1 newly diagnosed patients in 2005-2010, TDR was found in seven patients (4.7%). The prevalence of TDR to nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors, and protease inhibitors was 2% (3/150), 2% (3/150), and 0.7% (1/150), respectively. The majority of patients were infected with subtype B (134/150, 89%), while subtype A was detected in 6.0% (9/150), subtype D in 1.3% (2/150), and subtype G and CRF02_AG in 0.7% (one patient each). Three of 150 sequences could not be typed. Infection with subtype B was found to be significantly associated with male gender, Slovenia being reported as the country of the patient's nationality and origin of the virus, CDC class A, mode of transmission with homosexual/bisexual contact, sex with an anonymous person, and a higher CD4(+) count. Among patients carrying the subtype B virus, an MSM transmission route was reported in 87% of patients. Although the prevalence of TDR in Slovenia is still below the European average, active surveillance should be continued, especially among MSM, the most vulnerable population for HIV-1 infection in this part of Europe.

A 70-year-old patient was admitted to the Department of Oral Medicine for multiple oral ulcerations on the left buccal mucosa, around 0.5 cm in diameter, as well as on the gingiva. Otherwise, the patient suffered from chronic lymphocytic leukemia, hypogammaglobulinemia, chronic renal insufficiency, with complete afunction of the right kidney, asthma, hypertension, gastritis and prostate hyperplasia. Differential diagnosis of oral ulcerations included drug induced oral ulcerations, paraneoplastic pemphigus, viral ulcerations (cytomegalovirus, herpes simplex viruses), fungal ulcerations (candidiasis, aspergillosis, histoplasmosis, cryptococcosis) and bacterial ulcerations, as well as neutropenic ulcers. One of the possible explanations was that the lesions were due to the use of drugs, the more so as oral lesions evolved when the doses of allopurinol and chlorambucil were increased, and subsided when the doses of both drugs were decreased. However, we could not establish for sure whether the lesions were due to allopurinol or chlorambucil. According to literature data, allopurinol is one of the most frequent drugs known to induce skin adverse reactions, therefore we assumed that it was the culprit drug. Unfortunately, several weeks later the patient died from sepsis, pneumonia with respiratory insufficiency and multiorgan failure., Bolesnik u dobi od 70 godina upućen je na Zavod za oralnu medicinu zbog nekoliko oralnih ulceracija na lijevoj obraznoj sluznici veličine oko pola centimetra u promjeru te na gingivi. Inače je bolovao od kronične limfocitne leukemije, hipogamaglobulinemije, kronične bubrežne insufi cijencije s potpunom afunkcijom desnog bubrega, astme, hipertenzije, gastritisa i hiperplazije prostate. Diferencijalna dijagnoza oralnih ulceracija uključivala je oralne ulceracije uslijed uzimanja lijekova, paraneoplastični pemfi gus, virusne ulceracije (citomegalovirus, herpes simpleks viruse), gljivične ulceracije (kandidijaza, aspergiloza, histoplazmoza, kriptokokoza), bakterijske ulceracije te neutropenične ulceracije. Jedno od mogućih objašnjenja je bilo da su lezije vjerojatno nastale uslijed uzimanja lijekova, jer su se pojavile kada se doza alopurinola i klorambucila povećala, a počele su nestajati kada se doza tih lijekova smanjila. Ipak, nismo sigurni koji je od ova dva lijeka doveo do nastanka lezija, jer je nažalost nekoliko tjedana poslije bolesnik izdahnuo uslijed sepse, pneumonije i respiracijske insuficijencije te višeorganskog zatajenja.

Polimorfizam jednog nukleotida (engl. single nucleotide polymorphism, SNP) rs12979860 u blizini promotora gena za interleukin-28B (IL-28B) tj. genotipizacija IL-28B povezana je s prirodnim tijekom infekcije virusom hepatitisa C kao i s odgovorom na antivirusno i imunomodulacijsko liječenje kombinacijom pegiliranog interferona alfa i ribavirina (tzv. dvojna terapija). U eri direktno djelujućih antivirusnih lijekova genotip IL-28B više nije značajan prediktor ishoda liječenja no i dalje je važan parametar koji određuje prirodni tijek povijesti HCV infekcije. U zemljama koje nemaju univerzalni pristup novim terapijskim opcijama genotipizacija IL-28B i dalje se primjenjuje u pred-terapijskoj obradi bolesnika koji se liječe dvojnom terapijom. Cilj ovog rada bio je analizirati rezultate genotipizacije IL-28B (udio homozigota za T i C alel te heterozigota) u 595 bolesnika s kroničnim hepatitisom C koji su bili uključeni u kliničku skrb Zavoda za virusni hepatitis Klinike za infektivne bolesti "Dr. Fran Mihaljević" u Zagrebu od siječnja 2013. do prosinca 2015. g. SNP rs12979860 detektiran je metodom lančane reakcije polimerazom u stvarnom vremenu. Analiza SNP-a rs12979860 u genu za IL-28B pokazala je da su 335 od 595 (56,3%) ispitanika bili CT heterozigoti, 173 od 595 ispitanika (29,1%) C homozigoti i 87 od 595 ispitanika (14,6%) T homozigoti. Distribucija C i T alela u SNP-u rs12979860 gena za IL-28B sukladni su literaturnim podatcima analize prevalencije ovih alela u osoba bijele rase europskog porijekla., Single nucleotide polymorphism (SNP) rs12979860 in the promotor region of the gene coding for interleukin-28B (IL-28B) is associated with the natural history of hepatitis C virus (HCV) infection and response to antiviral and immunomodulatory therapy with combination of pegylated interferon alpha and ribavirin (dual therapy). Despite the fact that the predictive role IL-28B genotyping is decreasing in the era of highly successful direct acting antiviral (DAA)-based treatment, it remains to be useful as a tool for predicting a natural course of HCV infection. In resource-limited settings with restricted access to new treatment options IL-28B polymorphism typing is still a part of pre-treatment workup before admission of dual therapy. The aim of this study was to analyse the results of IL-28B genotyping (percentages of C and T homozygotes and heterozygotes) for 595 Caucasian patients with chronic hepatitis C receiving clinical care at the Department of Viral Hepatitis of the University Hospital for Infectious Diseases, Zagreb and Croatian Reference Center for Viral Hepatitis since January 2013 till December 2015. Detection of single nucleotide polymorphism (SNP) rs12979860 was determined by using a real-time PCR test. Analysis of rs12979860 SNP in the IL-28B gene showed that 335 of 595 (56.3%) patients were CT heterozygotes, followed by C homozygotes (173 of 595 patients, 29.1%) whereas T homozygotes were the least frequent (87 of 595 patients, 14.6%). The patterns of IL-28 polymorphism in Croatian patients with chronic hepatitis C is in accordance with other results for Caucasians.

HIV-positive individuals that have a detected transmitted drug resistance (TDR) at baseline have a higher risk of virological failure with antiretroviral therapy (ART). This study offers an update on the prevalence of TDR in Slovenia, looks for onward transmission of TDR, and reassesses the need for baseline drug resistance testing. Blinded questionnaires and partial pol sequences were obtained from 54.5% (168/308) of all of the patients diagnosed with HIV-1 from 2011 to 2016. Subtype B was detected in 82.7% (139/168) of patients, followed by subtype A (8.3%), subtype C (2.4%), and CRF01_AE (1.8%). Surveillance drug resistance mutations (SDRMs) were found in four individuals (2.4%), all of them men who have sex with men (MSM) and infected with subtype B. K103N was detected in two patients and T68D and T215D in one person each, corresponding to a prevalence of 0%, 1.2%, and 1.2% of TDR to protease inhibitors (PIs), nucleoside reverse transcriptase inhibitors (NRTIs), and non-NRTIs (NNRTIs), respectively. The impact of mutations on drug susceptibility was found to be most pronounced for NNRTIs. No forward spread of TDR within the country was observed; however, phylogenetic analysis revealed several new introductions of HIV into Slovenia in recent years, possibly due to increased risky behavior by MSM. This was indirectly confirmed by a substantial increase in syphilis cases and HIV-1 non-B subtypes during the study period. A drug-resistant HIV variant with good transmission fitness is thus more likely to be imported into Slovenia in the near future, and so TDR should be closely monitored.

Prometryne is a methylthio-s-triazine herbicide. Significant traces are documented in environment, mainly waters, soil and plants used for nutrition. The aim of this study was to estimate prometryne immunotoxic properties through induction of apoptotic and/or necrotic changes in thymocytes, splenocytes and lymph node cells after repeated subchronical exposure. Three different doses of prometryne (185, 375, 555mgkg(-1)) were applied per os every 48h, over 28 days. Flow cytometry assay (annexinV-FITC and PI) was conducted to record apoptotic and necrotic damage. In the spleen significant changes in the percentage of apoptotic cells were not detected between treated and control groups respectively. In thymus and lymph node, within the lowest dose group (185mgkg(-)1), an increase in percentage of early apoptosis without any significant increase in necrosis was detected. Medium (375mgkg(-1)) as well as high dose triggered increase in late apoptosis in lymph node while in thymus; late apoptosis was increased only in animals exposed to the highest dose (555mgkg(-1)). The highest applied dose, in thymus and lymph node respectively, caused a general decrease in percentage of vital cells in favour of marked increase of percentages of all types of dying cells (apoptotic, late apoptotic/early necrotic and necrotic). Prometryne caused disbalance in major organs of immune system, markedly lymph nodes and thymus, by induction of early apoptotic changes in dose/time specific manner.

OBJECTIVES: The aim of our study was to determine whether cerebrospinal fluid (CSF) of patients with tick-borne encephalitis (TBE) contains CXCL10, CXCL11, p40 subunit of interleukin-12 (IL-12)/IL-23, IL-18 and IL-15. We compared serum and CSF concentrations of CXCL10 and analysed the possible concentration gradient of this chemokine between the periphery and central nervous system. ----- MATERIALS AND METHODS: The study enrolled 19 TBE patients and 10 patients with non-inflammatory neurological diseases. ----- RESULTS: CSF of TBE patients contained CXCL10 (median 217 pg/ml), CXCL11 (8.3 pg/ml), p40 subunit of IL-12/IL-23 (38.9 pg/ml), IL-18 (30.1 pg/ml) and IL-15 (5.9 pg/ml). CXCL10 in the CSF of TBE patients was higher compared with serum (median 62 pg/ml, P < 0.001). ----- CONCLUSION: CSF of TBE patients contains CXCL10, CXCL11, p40 subunit of IL-12/IL-23, IL-18 and IL-15. Increased CXCL10 concentration in CSF suggests a role for this chemokine in the recruitment of CXCR3-expressing T-cells into the CSF of TBE patients.

Human papillomavirus type 6 (HPV6) is the major etiological agent of anogenital warts and laryngeal papillomas and has been included in both the quadrivalent and nonavalent prophylactic HPV vaccines. This study investigated the global genomic diversity of HPV6, using 724 isolates and 190 complete genomes from six continents, and the association of HPV6 genomic variants with geographical location, anatomical site of infection/disease, and gender. Initially, a 2,800-bp E5a-E5b-L1-LCR fragment was sequenced from 492/530 (92.8%) HPV6-positive samples collected for this study. Among them, 130 exhibited at least one single nucleotide polymorphism (SNP), indel, or amino acid change in the E5a-E5b-L1-LCR fragment and were sequenced in full. A global alignment and maximum likelihood tree of 190 complete HPV6 genomes (130 fully sequenced in this study and 60 obtained from sequence repositories) revealed two variant lineages, A and B, and five B sublineages: B1, B2, B3, B4, and B5. HPV6 (sub)lineage-specific SNPs and a 960-bp representative region for whole-genome-based phylogenetic clustering within the L2 open reading frame were identified. Multivariate logistic regression analysis revealed that lineage B predominated globally. Sublineage B3 was more common in Africa and North and South America, and lineage A was more common in Asia. Sublineages B1 and B3 were associated with anogenital infections, indicating a potential lesion-specific predilection of some HPV6 sublineages. Females had higher odds for infection with sublineage B3 than males. In conclusion, a global HPV6 phylogenetic analysis revealed the existence of two variant lineages and five sublineages, showing some degree of ethnogeographic, gender, and/or disease predilection in their distribution. Fil: Jelen, Mateja M. University of Ljubljana; Eslovenia Fil: Chen, Zigui. Albert Einstein College of Medicine. New York; Estados Unidos Fil: Kocjan, Botjan J.. University of Ljubljana; Eslovenia Fil: Burt, Felicity J.. University of the Free State. Bloemfontein; Sudáfrica Fil: Chan, Paul K.S.. Chinese University of Hong Kong; China Fil: Chouhy, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina Fil: Combrinck, Catharina E.. University of the Free State. Bloemfontein; Sudáfrica Fil: Coutlee, Francois. Hôpital Notre-Dame-Pavillon Deschamps. Montréal; Canadá Fil: Estrade, Christine. Lausanne University Hospital; Suiza Fil: Ferenczy, Alex. McGill University and Jewish General Hospital. Montreal; Canadá Fil: Fiander, Alison. Institute of Cancer and Genetics. Cardiff; Reino Unido Fil: Franco, Eduardo L.. McGill University. Montreal; Canadá Fil: Garland, Suzanne M.. The Royal Women's Hospital. Parkville; Australia. The University of Melbourne; Australia. Murdoch Children's Research Institute. Victoria; Australia Fil: Giri, Adriana Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina Fil: González, Joaquín Víctor. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud; Argentina Fil: Gröning, Arndt. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; Alemania Fil: Heidrich, Kerstin. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; Alemania Fil: Hibbitts, Sam. Institute of Cancer and Genetics. Cardiff; Reino Unido Fil: Hosnjak, Lea. University of Ljubljana; Eslovenia Fil: Luk, Tommy N.M. The Chinese University of Hong Kong; China. Hong Kong Special Administrative Region; China Fil: Marinic, Karina. Hospital Dr. Julio C. Perrando. Chaco; Argentina Fil: Matsukura, Toshihiko. National Institute of Health. Tokyo; Japón Fil: Neumann, Anna. The University of Melbourne; Australia. Medizinisches Versorgungszentrum wagnerstibbe für Laboratoriumsmedizin und Pathologie GmbH. Hannover; Alemania Fil: Oštrbenk, Anja. University of Ljubljana; Eslovenia Fil: Picconi, Maria Alejandra. Dirección Nacional de Instituto de Investigación. Administración Nacional de Laboratorio e Instituto de Salud; Argentina Fil: Richardson, Harriet. Queen's University. Ontario; Canadá Fil: Sagadin, Martin. University of Ljubljana; Eslovenia Fil: Sahli, Roland. Lausanne University Hospital; Suiza Fil: Seedat, Riaz Y.. University of the Free State. Bloemfontein; Sudáfrica Fil: Seme, Katja. University of Ljubljana; Eslovenia Fil: Severini, Alberto. Public Health Agency of Canada. Winnipeg; Canadá Fil: Sinchi, Jessica L. Hospital Dr. Julio C. Perrando. Chaco; Argentina Fil: Smahelova, Jana. Institute of Hematology and Blood Transfusion. Praga; República Checa Fil: Tabrizi, Sepehr N.. The University of Melbourne; Australia. Murdoch Children's Research Institute. Victoria; Australia. The Royal Women's Hospital. Parkville; Australia Fil: Tachezy, Ruth. Institute of Hematology and Blood Transfusion. Praga; República Checa Fil: Tohme, Sarah. Public Health Agency of Canada. Winnipeg; Canadá Fil: Uloza, Virgilijus. Lithuanian University of Health Sciences. Kaunas; Lituania Fil: Vitkauskiene, Astra. Lithuanian University of Health Sciences. Kaunas; Lituania Fil: Wong, Yong Wee. Universti Kebangsaan Malaysia. Bangi; Malasia Fil: Lepeja, Snježana Židovec. University Hospital for Infectious Diseases Dr. Fran Mihaljević. Zagreb; Croacia Fil: Burk, Robert D.. Albert Einstein College of Medicine.New York; Estados Unidos Fil: Poljak, Mario. Albert Einstein College of Medicine. New York; Estados Unidos

Opisali smo glavne novosti u dijagnostici hepatitisa B i C kao dio hrvatskih smjernica za dijagnostiku i liječenje virusnih hepatitisa 2013. Dijagnostika akutnog i kroničnog hepatitisa B započinje određivanjem HBsAg, anti-HBc i anti-HBs. Ostale serološke biljege hepatitisa B treba određivati tek u drugom koraku ako su nalazi HBsAg i/ili anti-HBc pozitivni. Pozitivan nalaz samo na anti-HBc potrebno je obvezno nadopuniti određivanjem HBV DNK. Kvantitativno određivanje HBsAg treba se koristiti komplementarno s određivanjem HBV DNK za: (i) razlikovanje inaktivnog nositelja HBsAg od aktivnog kroničnog HBeAg-negativnog hepatitisa B u bolesnika sa HBV DNK nižom od 2.000 IU/mL te za (ii) praćenje tijeka liječenja kroničnog hepatitisa B s pegiliranim interferonom-alfa. Metoda PCR-a u stvarnom vremenu ostaje i dalje metoda izbora za detekciju i kvantifikaciju HBV DNK. Testiranje za HCV započinje određivanjem specifičnih protutijela probirnim enzimskim imunotestovima ili brzim testovima. Sve osobe s pozitivnim probirnim anti-HCV testom treba testirati na prisutnost HCV RNK ili antigena virusne kapside. Potvrdne anti-HCV testove treba koristiti samo kao dodatne testove koji će potvrditi ili isključiti značenje reaktivnih rezultata probirnih enzimskih imunotestova u osoba koje su HCV RNK negativne. U praćenju virusne kinetike tijekom trojne terapije hepatitisa C preporučuje se korištenje molekularnih testova s identičnom donjom granicom detekcije (LLOD) te donjom granicom kvantifikacije. Određivanje rezistencije HCV-a na inhibitore proteaze nije dio obveznog dijagnostičkog praćenja bolesnika liječenih s trojnom terapijom. Ne postoje dostatni dokazi o potrebi uvođenja subtipizacije HCV-a u obveznu predterapijsku obradu bolesnika s kroničnim hepatitisom C. Genotip IL-28 je važan prediktor SVR-a u bolesnika liječenih kombinacijom IFN i ribavirina kao i u bolesnika zaraženih genotipom 1 liječenih trojnom terapijom. Genotipizacija IL-28B preporučuje se u predterapijskoj obradi bolesnika s kroničnim hepatitisom C. Posebno je značajan dijagnostički parametar u terapijski-naivnih bolesnika s kroničnim hepatitisom C prilikom dvojbe dvojna vs. trojna terapija., The 2013 Update of the Croatian Guidelines for the Diagnosis and Treatment of Viral Hepatitis summarizes recent developments in the diagnosis of hepatitis B and C. Determination of HBsAg, anti-HBc and anti-HBs is the initial step in the diagnostic workup of acute and chronic hepatitis B. Other hepatitis B serologic markers should be analyzed in the second stage of the diagnostic workup in HBsAg and/or anti-HBc positive patients. A positive anti-HBc finding should be followed by HBV DNA quantification. HBsAg quantification is complimentary to the HBV DNA quantification and is used: (i) to differentiate between inactive HBsAg carriers and active chronic HBeAg-negative hepatitis B in patients with HBV DNA

Hrvatske konsenzus konferencije o virusnim hepatitisima održane su 2005. i 2009. g. (1). S obzirom na brojne nove spoznaje o epidemiologiji, dijagnostici i liječenju virusnih hepatitisa (poglavito kroničnog hepatitisa C genotipa 1) u protekle četiri godine, 28. veljače 2013. g. održana je nova Hrvatska konsensus konferencija o virusnim hepatitisima u Zagrebu. Sažeti tekst ove Hrvatske konsenzus konferencije o virusnim hepatitisima sadrži prikaz novih spoznaja o epidemiologiji virusnih hepatitisa, serološkoj i mo¬lekularnoj dijagnostici virusnih hepatitisa, određivanju polimorfizma promotora gena za IL-28, procjeni stadija fibroze, algoritmu dijagnostičkog praćenja bolesnika, liječenju kroničnog hepatitisa C (genotipovi 1-6) i hepatitisa B, liječenju specijalnih populacija (djeca, bolesnici na dijalizi, bolesnici liječeni transplantacijom, osobe s HIV/HCV koinfekcijom) i nuspojavama liječenja., Croatian Consensus Conferences on Viral Hepatitis took place in 2005 and 2009. Considering the numerous novel concepts on the epidemiology, diagnosis and management of viral hepatitis (chronic hepatitis C genotype 1 in particular) that have emerged in the past four years, a new Croatian Consensus Conference on Viral Hepatitis was held in Zagreb on February 28, 2013. The abridged text of the Croatian Consensus Conference on Viral Hepatitis 2013 presents the new concepts on the epidemiology of viral hepatitis, serologic and molecular diagnosis of viral hepatitis, determination of the IL-28 gene promoter polymorphism, fi¬brosis grading, algorithm for patient diagnostic follow up, treatment of chronic hepatitis C (genotypes 1-6) and hepatitis B, treat¬ment of special populations (children, dialysis patients, transplanted patients, individuals with HIV/HCV co-infection), and therapy side effects.

Trajni virusološki odgovor nakon 48 tjedana liječenja kombinacijom pegiliranog interferona alfa-2 i ribavirina u bolesnika s kroničnim hepatitisom C inficiranih genotipom 1 postiže se u oko 55% bolesnika. Prethodne studije pokazale su da je kraća, 24-tjedna terapija u bolesnika s niskom početnom viremijom (, About 55% of patients with genotype 1 chronic hepatitis C infection have sustained response to 48-week therapy with pegylated interferon alfa-2 and ribavirin. Previous studies have shown that patients with low pretreatment viremia (< 600 000 IU/mL) can be treated for 24 weeks with similar efficacy as with 48-week therapy, when early viral kinetic is measured. Our study showed that patients with low pretreatment viral load (< 600 000 IU/mL) are very heterogenous group and sustained viral response to pegylated interferon alfa-2 and ribavirin therapy is not better than in high viral load group (low viremia patients SVR 53.2% (n = 94), high viremia patients SVR 50.8% (n = 122)). Pretreatment low viral load patients that did not achieve SVR (n = 44) had a significantly higher fibrosis stage (non-SVR fibrosis stage 3.47 Ishak score vs. SVR+ fibrosis stage 2.94; p = 0,004), and they were significantly older (non-SVR patients 46.4 years, SVR+ patients 39.6 years, p = 0.008). Low initial viremia patients with cirrhosis (n = 6) did not achieve SVR at all, while among patients with fibrosis stage 4/6, 52 % achieved SVR. Our results suggest that apart from viral load at the beginning of treatment, fibrosis stage is also a predicting factor for therapy outcome, particularly in older patients and patients with cirrhosis.

Cilj ovog istraživanja bio je odrediti heterogenost podtipova HIV-a te analizirati primarnu rezistenciju virusa na antiretrovirusne lijekove u 30 prethodno neliječenih osoba koje su uključene u kliničku skrb Referentnog centra za dijagnostiku i liječenje HIV/AIDS-a tijekom 2013. g. Sekvenciranjem pol gena HIV-a te primjenom bioinformatičkih algoritama određeni su podtipovi virusa te detektirane mutacije povezane s primarnom rezistencijom na antiretrovirusne lijekove. Većina ispitanika (86,7%) bila je zaražena podtipom B. Otkriveno je 12 mutacija povezanih s primarnom rezistencijom HIV-a na antiretrovirusne lijekove, tj. 9 mutacija povezanih s primarnom rezistencijom na nukleozidne analoge inhibitore reverzne transkriptaze (NRTI) te 3 mutacije povezane s rezistencijom na nenukleozidne inihibitore reverzne transkriptaze (NNRTI). Primarna rezistencija na dvije klase antiretrovirusnih lijekova (NRTI i NNRTI) otkrivena je u jednog ispitanika. Najzastupljenija mutacija povezana s primarnom rezistencijom virusa na lijekove bila je T215S (rezistencija na NRTI). Mutacije značajne za nastanak primarne rezistencije detektirane su u 4 osobe zaražene podtipom B, jedne osobe zaražene podtipom A1 te jedne osobe zaražene rekombinantom podtipova A1 i B. U ovom je istraživanju po prvi puta dokazana pojava primarne rezistencije HIV-a na antiretrovirusne lijekove u osoba zaraženih non-B podtipovima virusa iz Hrvatske., The aim of this study was to analyse HIV subtype diversity as well as primary resistance to antiretroviral drugs in 30 treatment-naive individuals entering clinical care at the National Refence Center for HIV/AIDS during 2013. Sequencing the pol region of the viral genome and bioinformatic algorithms were used to determine HIV subtypes and to analyse transmitted drug resistance mutations. The majority of individuals (86,7 %) were infected with HIV-1 subtype B. Twelve mutations associated with primary resistance to antiretroviral drugs were detected in the study: 9 mutations conferring resistance to NRTI (nucleotide reverse-transcriptase inhibitors) and 3 mutations associated with resistance to NNRTI (non-nucleotide reverse-transcriptase inhibitors). Primary resistance to two antiretroviral drug classes (NRTI and NNRTI) were detected in one individual. The most frequent mutation was T1215S associated with primary resistance to NRTI. Drug resistance mutations were detected in 4 subtype B and 2 non-B HIV-infected individuals. This study showed, for the first time, the presence of mutations associated with primary resistance of HIV to antiretroviral drugs in individuals infected with non-B subtypes in Croatia.

Background We assessed the feasibility of collecting urine samples for testing on genital Chlamydia trachomatis infection in a population-based survey, and prevalence of this infection among young people aged 18-25 in Croatia. In Croatia, as in the other countries of Eastern Europe, there is a lack of data on prevalence of C trachomatis in the general population, including young adults. Methods We sampled participants using a nationally representative, multi-stage stratified probability sample of young men and women. Detection of C trachomatis DNA in urine samples was performed by using a real-time PCR assay COBAS® TaqMan® CT Test, v2.0. Results Overall, 1005 young adults participated in the behavioural part of the survey, and 27.9% men and 37.5% women who were sexually experienced agreed to provide urine samples for testing on C trachomatis. Using multivariate analysis, women were significantly more likely to provide urine samples than men (aOR = 1.53, 95% CI 1.14-2.06) as were those who reported no condom use at last intercourse (aOR = 1.95, 95% CI 1.44-2.62). Prevalence of C trachomatis infection among those who were sexually experienced was 7.3% in men and 5.3% in women. Conclusions Population-based surveys that use probabilistic sampling are a feasible way to obtain population estimates of C trachomatis prevalence among young adults in Croatia, but it is challenging to obtain an adequate response rate. The prevalence of C trachomatis among young adults in Croatia found in this study was higher than that found in other European countries with similar survey response rates.

Aim. To examine the risk factors, comorbidity, severity of liver disease, treatment course, and outcome in Croatian war veterans with chronic hepatitis C, especially those suffering from posttraumatic stress disorder (PTSD). ----- Methods. We collected medical records of 170 adult men diagnosed with chronic hepatitis C who started treatment with a combination of pegylated interferon-alpha and ribavirin between January 2003 and June 2009 at the Croatian Reference Centre for Viral Hepatitis. ----- Results. Participants' mean age was 43±9 years. Among 170 participants, there were 37 war veterans (22%). The main risk factor in veteran patients were operative procedures with transfusions (46% vs 5% in non-veterans; P

BACKGROUND: Lymphocyte migration from the blood into the CNS is mediated by chemokines and chemokine receptors. Chemokines CXCL10 and CXCL11 are important for the recruitment of CXCR3-expressing Th1 lymphocytes to the site of inflammation. ----- AIMS: To determine the concentrations of CXCL10 and CXCL11 in the CSF and plasma of children with enteroviral aseptic meningitis (EV AM) and controls and the contribution of these chemokines to the chemokine concentration gradient between the periphery and the CNS. ----- METHODS: The study included 26 pediatric patients with EV AM and 16 controls in whom CNS infection is excluded by negative CSF examination. Chemokines were quantified by using enzyme immunoassay. Etiological diagnosis of EV AM was based on the detection of enteroviral RNA in the CSF using real-time PCR. ----- RESULTS: CXCL10 (median 12 725 pg/ml) and CXCL11 (median 187 pg/ml) concentrations in CSF of patients with meningitis were significantly higher compared to plasma (median 173 pg/ml and median 110 pg/ml; p < 0.001, p = 0.026 respectively). CXCL10 concentrations in the CSF (median 198 pg/ml) and plasma of controls (median 124 pg/ml) were not significantly different (p = 0.642). CXCL11 concentrations in the CSF of controls (median 89 pg/ml) were significantly lower compared with plasma (median 139 pg/ml, p = 0.004). Chemokine concentration gradient was not influenced by pleocytosis, nor dependent on cytologic CSF formula or the presence of proteinorrachia. ----- CONCLUSION: CXCL10 and CXCL11 concentration gradient between the CSF and plasma in children with EV AM suggests an important role of these chemokines in the T-cells recruitment into the CNS and local immunoreaction.

Humani papilomavirusi (HPV) su heterogena skupina virusa koja se tijekom evolucije izvrsno prilagodila replikaciji u stanicama epitela. Perzistentna infekcija humanim papilomavirusima obvezan je etiološki kofaktor u nastanku benignih i malignih novotvorina pločastog epitela. Prepoznavanje različitosti onokogenog potencijala pojedinih genotipova HPV-a potaknulo je brojna istraživanja kliničke značajnosti molekularne heterogenosti ove skupine virusa. Molekularna analiza HPV-a značajna je ne samo u temeljnim virusološkim istraživanjima već i u humanoj medicini s posebnim naglaskom na problematiku povezanosti HPV-a s određenim tipovima tumora, razvoj klinički vrijednih dijagnostičkih testova, procjenu rizika od nastanka malignih lezija, sastavljanje kliničkih postupnika za probir, praćenje i liječenje zaraženih, razvoj cjepiva, analizu "pre-cjepne" distribucije genotipova, procjenu obuhvatnosti i učinkovitosti cjepiva i praćenje distribucije genotipova nakon uvođenja univerzalnog cijepljenja., Human papillomaviruses (HPV) are a heterogeneous group of viruses that have throughout their evolution adapted to replication in epithelial cells. Persistent infection with human papillomaviruses represents a necessary co-factor for the development of benign and malignant epithelial neoplasia. Different oncogenic potential of individual HPV genotypes encouraged numerous studies on clinical significance of the observed molecular heterogeneity of HPV. Molecular analysis of HPV is relevant not only for basic virological research but for human medicine as well with particular emphasis on association between HPV and various types of human tumors, development of clinically validated diagnostic assays, assessment of risk for progression of cervical lesions, vaccine design and development, analysis of pre-vaccination distribution of HPV genotypes, analysis of vaccine coverage and efficacy as well as monitoring of possible genotype replacement following introduction of universal HPV vaccination.

Toxoplasma gondii jedan je od glavnih oportunistički patogena u bolesnika zaraženih HIV-om. Prevalencija zaraze parazitom T. gondii u HIV-bolesnika u Hrvatskoj do sada nije poznata. Cilj rada bio je utvrditi seroprevalenciju toksoplazmoze u HIV-bolesnika u Hrvatskoj. Uključeno je 166 HIV-bolesnika i 219 dobrovoljnih davatelja krvi (DDK) kao zdrava kontrola. Enzimskim imunotestom određena su protutijela IgM, IgG i IgA na T. gondii. Seroprevalencija toksoplazmoze u HIV-bolesnika iznosi 51,8 %, a u DDK 52,5 %. Protutijela IgM bila su reaktivna samo u 2 HIV-bolesnika i 2 DDK, a IgA u 2 HIV-bolesnika i 1 DDK. Prema rizičnom ponašanju za HIV-infekciju utvrđeno je da muškarci koji imaju spolne odnose s drugim muškarcima imaju statistički značajno višu prevalenciju (66,7 %) i 2,4 puta veću šansu za toksoplazmozu nego drugi HIV-bolesnici (OR 2,37; 95 % IP 1,12–5,13). Rezultati su analizirani prema dobi, spolu, geografskom podrijetlu i za HIV-bolesnike broju CD4+ limfocita T i nisu nađene statistički značajne razlike ni za jedan parametar. Ovo je prvo istraživanje u kojem je utvrđena seroprevalencija toksoplazmoze u HIV-bolesnika u Hrvatskoj., Toxoplasma gondii is one of the main opportunistic agents in HIV-infected patients. The prevalence of T. gondii infection in HIV-infected patients in Croatia is unknown. The aim of the study was to determine the seroprevalence of toxoplasmosis in HIV-infected patients in Croatia. Included in the study were 166 HIV-infected patients and 219 blood donors (BD) as healthy control. Anti-toxoplasma IgM, IgG and IgA antibodies were determined with enzyme immunoassay respectively. Seroprevalence of toxoplasmosis in HIV-patients was 51.8 % and in BD 52.5 %. IgM antibodies were found in 2 HIV-patients and 2 BD, and IgA in 2 HIV-patients and 1 BD. According to HIV risk behaviour men who have sex with other men had significantly higher prevalence rate (66.7 %) and 2.4 higher risk for toxoplasma infection than other HIV-infected patients (OR 2.37; 95 % CI 1.12–5.13). Results were analyzed according to age, gender, geographic origin and for HIV-infected patients also according to CD4+ T-cell count number, and no statistically significant differences were found. This is the first study that presents seroprevalence of toxoplasmosis in HIV-infected patients in Croatia.

A genotyping study of 285 Hybrid Capture 2 low-risk probe cocktail-positive specimens showed cross-reactivity with several untargeted human papillomavirus genotypes. Cross-reactivity was often clinically beneficial due to the detection of untargeted low-risk genotypes. A total of 8.4% of positive results, usually weak, were due to cross-reactivity with high-risk genotypes. Establishment of a gray zone is recommended.

Virusološka dijagnostika HCV infekcije temelji se na primjeni standardiziranih seroloških i molekularnih testova. U molekularnoj dijagnostici HCV infekcije koriste se vrlo osjetljivi testovi za kvantifikaciju HCV RNK i genotipizaciju HCV-a. Genotip HCV-a važan je čimbenik pred-terapijske obrade bolesnika koji određuje trajanje liječenja, dozu ribavirina a omogućuje i procjenu vjerojatnosti uspjeha liječenja. Međutim, detekcija i/ili kvantifikacija HCV RNK u serumu preporučuje se za dokazivanje aktivne virusne replikacije u sklopu pred-terapijske obrade bolesnika kao i za praćenje virusološkog odgovora tijekom liječenja. Rezultati praćenja distribucije genotipova HCV-a u Referentnom centru za virusni hepatitis Ministarstva zdravstva R. Hrvatske pokazuju da je oko 60 % bolesnika s kroničnim hepatitisom C zaraženo genotipom 1 HCV-a, 36 % genotipom 3a, a svega 4 % bolesnika genotipovima 2 i 4. Zlatni standard liječenja kroničnog hepatitisa C (KHC) posljednjih sedam godina je kombinacija pegiliranog interferona alfa (PEG IFN) i ribavirina. Međutim, standardna terapija ne dovodi do izliječenja u 50 % bolesnika s genotipom 1, te oko 30 % bolesnika s genotipom 3a. Stoga se suvremeno liječenje bolesnika s kroničnim hepatitisom C temelji se na individualnom pristupu koji uključuje prilagodbu algoritma liječenja genotipu virusa, viremiji, stadiju fibroze te praćenju virusne kinetike. Modifikacija algoritma liječenja u bolesnika s genotipom 1 i sporim virusološkim odgovorom uključuje produljenje liječenja na 72 tjedna dok se liječenje bolesnika zaraženih genotipom 3a koji imaju nisku viremiju, a postignu brzi virusološki odgovor, liječenje može skratiti na 16 tjedana. Bolesnike s visokim stadijem fibroze (obilna vezivna septa) nije uputno liječiti prema visini viremije, jer u toj skupini bolesnika niska viremija ne predstavlja prediktor trajnog virusološkog odgovora. Bolesnike s akutnim hepatitisom C treba liječiti monoterapijom pegiliranim interferonom alfa u trajanju od 24 tjedna., Virological diagnostics of HCV infection is based on standardized serological and molecular assays. Molecular diagnostics of HCV infection includes assays for HCV RNK detection and/or quantification as well as genotyping assays. HCV genotype is an important parameter in the pre-treatment diagnostic workup that determines treatment duration, ribavirin dosage and represents an excellent predictor of treatment success. Detection and/or quantification of HCV RNK in the serum provides direct evidence of active viral replication in patients with chronic hepatitis C and enables determination of virological response during treatment. According to the Croatian Reference Center for Viral Hepatitis, 60 % of patients with chronic hepatitis C in Croatia are infected with genotype 1, 36 % of patients with genotype 3a and only 4 % of patients with genotypes 2 and 4. Standard treatment regimen for chronic hepatitis C over the past seven years is a combination of pegylated interferon alpha (PEG IFN) and ribavirin (guanosine analogue). However, standard treatment regimen based on the combination of PEG IFN and ribavirin for 48 weeks fails to achieve sustained viral response in 50 % of patients with genotype 1. Furthermore, 25 % of patients infected with genotype 3a will fail to respond to a standard 24 week regiment. Up-to date treatment of chronic hepatitis C should be individualized (treatment guided) according to the genotype, liver fibrosis, early viral kinetics and viremia. Modification of treatment algorithm in patients with genotype 1 who are late responders to treatment should include prolonged treatment (72 weeks). In patients infected with genotypes 2 and 3 and low viremia who are rapid responders, therapy can be shortened to 16 weeks. Patients with higher fibrosis rates (fibrotic septa) should not be treated according to viremia because in those patients viremia does not correlate with the ability to achieve sustained viral response. Meta-analysis of recent clinical trials on the treatment of acute hepatitis C showed that monotherapy with PEG IFN for 24 weeks is the suggested treatment regimen for this group of patients.

In this report, the authors present a detailed immunological and virological assessment of an immunocompetent 17-year-old Caucasian male with a fatal Epstein-Barr virus (EBV) infectious mononucleosis presenting with meningoencephalitis and hemophagocytic syndrome. The patient with serologically confirmed EBV infectious mononucleosis was admitted to the hospital because of 3 weeks' fever. Fine-needle aspiration of lymph nodes showed reactive hyperplasia with prominent hemophagocytosis. Percentages of intracellular interferon-gamma (IFN-gamma) in CD4(+) and CD8(+) T cells in the peripheral blood progressively increased during the course of disease (10.2% and 8.5% on day 35 ; 30.1% and 53.2% on day 44 ; 42.2% and 75.2% on day 50 ; 36.1% and 50.6% on day 59, respectively). On day 50, the patient developed meningoencephalitis. Brain computed tomography (CT) was normal. Brain magnetic resonance imaging (MRI) showed multifocal inflammatory lesions in frontal and temporal cortex of the right hemisphere as well as severe perivascular inflammatory reaction. The patient was treated with steroids, cyclosporin A, and methotrexate intratecally. Following treatment, EBV viremia in the blood and cerebrospinal fluid (CSF) decreased from pretreatment values (54, 490 copies of EBV DNA/ml and 39, 500 copies/ml, respectively) to 8715 copies/ml in the blood and 14, 690 in the CSF. Despite treatment, the patient remained unconscious and died of sepsis and pneumonia 3 months after initial symptoms. Immunohistochemical staining showed the presence of EBV in both perivascular infiltrates and grey matter. Enhanced Th1 response as shown by high levels of IFN-gamma in peripheral blood lymphocytes may be a predictor of severe complications during acute EBV infection. Early implementation of immunosuppressive therapy in these patients should be considered.

Rad opisuje prve dostupne podatke o molekularnoj dijagnostici sepse primjenom multiplex real-time PCR-a u transplantiranih bolesnika.

We investigated the distribution of memory (CD45RO+) and naive (CD45RA+CD62L+) CD4+ T-cells as well as CD8+ T-cells and total T-cells in the CSF of children with aseptic meningitis following measles-mumps-rubella (MMW) vaccination and those with enteroviral meningitis. Flow cytometric analysis of CSF cells was performed in 12 children with MMR vaccine-associated meningitis and 11 children with enteroviral meningitis. Percentages of total T-cells, CD4+ and CD8+ T-cells and monocytes in CSF of patients from the two groups were not significantly different. The majority of CD4+ T-cells in the CSF of both patient groups were of memory phenotype. Percentages of CSF naive CD4+ T-cells were increased in children with aseptic meningitis following MMR vaccination. Further studies focused on the more detailed immunophenotyping of CSF cells are needed to fully establish the usefulness of flow cytometry in the diagnostic workup of inflammatory CNS diseases in children.

The aim of this study was to evaluate the performance of a molecular assay for quantitation of Epstein-Barr virus (EBV) DNA based on real-time PCR, and to determine EBV DNA levels in EDTA whole blood samples derived from different groups of patients. Following a manual DNA extraction protocol, real-time PCR was performed using the LightCycler EBV Quantification Kit, which demonstrated sufficient accuracy and linearity. Coefficients of variations were found to be between 6 and 42% and 5 and 34%, respectively, for inter-assay and intra-assay variations. In clinical specimens, EBV DNA was detected in all patients with acute EBV infection (n=34), in one of 25 adults with past EBV infection, in 16 out of 25 (64%) anti-HIV antibody positive persons, in 10 out of 25 (40%) solid organ transplant recipients, and in none of the 23 infants without history of EBV infection. When EBV DNA levels in positive specimens were compared between different groups, statistically significant differences were not found. The LightCycler EBV Quantification Kit was found to be useful for determination of EBV DNA levels in EDTA whole blood.