Polypharmacy and potentially inappropriate prescribing of benzodiazepines in older nursing home residents.

INTRODUCTION: Previous research has raised concerns about high prevalence of drug-related problems, polypharmacy and inappropriate benzodiazepine prescribing in nursing homes (NHs) and confirmed lack of studies from Central and South-Eastern Europe. The aim of our study was to determine the prevalence and characteristics of polypharmacy, hyperpolypharmacy and inappropriate benzodiazepine prescribing in NH residents in Croatia. METHODS: Data from 226 older NH residents from five Croatian NHs were collected using the InterRAI Long-Term Care Facilities assessment form. The prevalence and determinants of polypharmacy/hyperpolypharmacy and patterns of inappropriate benzodiazepine prescribing were documented. RESULTS: The prevalence of polypharmacy (49.6%) and hyperpolypharmacy (25.7%) among NH residents was high. In our study, 72.1% of NH residents were prescribed at least one psychotropic agent, 36.7% used 2-3 psychotropics and 6.6% used 4+ psychotropics. Among benzodiazepine users (55.8%), 28% of residents were prescribed benzodiazepines in higher than recommended geriatric doses, 75% used them for the long term and 48% were prescribed concomitant interacting medications. The odds of being prescribed polypharmacy/hyperpolypharmacy were significantly higher for older patients with polymorbidity (6+ disorders, proportional odds ratio (POR) = 19.8), type II diabetes (POR = 5.2), ischemic heart disease (POR = 4.6), higher frailty (Clinical Frailty Scale (CFS ≥5); POR = 4.3) and gastrointestinal problems (POR = 4.8). CONCLUSIONS: Our research underscores the persistent challenge of inappropriate medication use and drug-related harms among older NH residents, despite existing evidence and professional campaigns. Effective regulatory and policy interventions, including the implementation of geriatrician and clinical pharmacy services, are essential to address this critical issue and ensure optimal medication management for vulnerable NH populations.

The mediating role of depressive, anxiety, and physical symptoms on work ability index in employed women with breast cancer: a prospective study from Croatia.

AIM: To explore the relationship between the current work ability index (WAI) and depressive and anxiety symptoms in breast cancer (BC) patients and the role of depressive, anxiety, and physical symptoms in mediating this relationship. METHODS: This prospective study enrolled 83 employed women with BC. At baseline assessment (in the first three months following BC diagnosis) and follow-up assessment (one year after baseline), participants completed the WAI, Beck Depression Inventory-II, State-Trait Anxiety Inventory, and European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire with a breast cancer-specific module. Mediation analyses were conducted to explore the mechanism by which depressive, anxiety, and physical symptoms influenced the relationship between WAI and depressive and anxiety symptoms. RESULTS: WAI was negatively associated with depressive and anxiety symptoms. The effect of baseline depressive and trait anxiety symptoms on WAI at follow-up was mediated by both depressive and trait anxiety symptoms, as well as by physical symptoms at follow-up. The effect of baseline state anxiety symptoms on WAI at follow-up was mediated only by state anxiety symptoms at follow-up. CONCLUSIONS: Baseline depressive and anxiety symptoms affect WAI at follow-up not only through persisting depressive and anxiety symptoms observed at follow-up but also through physical symptoms at follow-up. This indicates that efforts aimed at improving psychological health may result in simultaneous improvements in both psychological and physical health, as well as the resulting WAI.

Impact of the number of therapy lines on survival in advanced gastric and esophagogastric adenocarcinoma - a real-world retrospective analysis from Croatia.

The aim of the study was to conduct a retrospective database analysis to understand the current treatment patterns and outcomes to plan potential improvements in therapy delivery and patient selection. The electronic patient medical records of 225 patients with advanced gastric and esophagogastric adenocarcinoma treated at two Croatian high-volume tertiary centers from January 2018 to December 2021 were analyzed. Patients ineligible for chemotherapy (66 of 291, 22.7%) due to poor general condition or co-morbidities were not included in the study. The median overall survival (OS) for the whole cohort was 11.0 months (95% confidence interval (CI) 9.7-12.0). Of the 225 patients who received first-line therapy, 47.6%, 16.9%, and 3.1% received second-, third-, and fourth-line therapy, respectively. Survival correlated significantly with the number of treatment lines received (p<0.001), with a median OS from diagnosis of 7.8 (95% CI 6.6-9.4), 12.0 (95% CI 10.0-14.0), and 20.0 months (95% CI 18.0-23.0) for patients receiving 1, 2, and ≥3 lines of treatment, respectively. This study confirmed the positive impact of the number of chemotherapy lines on OS. This highlights the importance of the ratio of patients receiving multiple lines of therapy as well as the availability of new and effective drugs in real-life clinical practice. The selection of optimal therapy for each patient in the first-line therapy is important because a significant number of patients do not receive second-line therapy.

Sex Ratio at Birth after Concurrent Events of Earthquakes and the COVID-19 Pandemic in Croatia.

The sex ratio at birth (ratio of males to females) has been known to be affected by exogenous shocks such as wars, pollution, natural catastrophes, economic crises, and others. Among these stressful events, both earthquakes and the COVID-19 pandemic have been reported to lower the sex ratio at birth. In this article, a rather unusual situation of two episodes of simultaneous events of COVID-19 lockdown and earthquakes approximately nine months apart (March and December of 2020) is investigated to assess whether they were associated with a bias in sex ratio at birth 3-5 months later (in utero loss) and 9 months later (loss at conception) in Croatia. The monthly time series of sex ratio at birth, total number of births, and total number of both male and female births from January 2010 to December 2021 were analyzed. Seasonally adjusted autoregressive moving-average models were used to estimate the functional form of the time series from January 2010 to February 2020. These results were used to predict the future values of the series until December 2021 and to compare them with the actual values. For all series used, there was no indication of deviation from the values predicted by the models, neither for 3-5 months nor for 9 months after the COVID-19 lockdown and earthquake events. The possible mechanisms of the absence of bias, such as the threshold of the stressful events and its localized reach, as well as the statistical methods employed, are discussed.

Prevalence of risk factors for development of cardiovascular diseases in urban and rural areas of Eastern Croatia: a cross-sectional study.

OBJECTIVE: Cardiovascular diseases are the leading global cause of death. Due to the disparities in cardiovascular risk factors between rural and urban populations, this study aims to assess the differences in the prevalence of risk factors in urban and rural areas of eastern Croatia. METHODS: The cross-sectional study included 280 participants (140 from urban and 140 from rural areas) registered at studied general practice offices. Methods included e-health records, questionnaire, physical examination methods, and blood sampling for laboratory tests. RESULTS: The most common risk factors among participants were elevated total cholesterol (83.6%), elevated LDL cholesterol (81.8%), increased body mass index (75.0%), increased waist-hip ratio (82.9%), increased waist circumference (63.2%), and arterial hypertension (70.1%). The rural participants had a significantly higher prevalence of arterial hypertension (p = 0.023), increased body mass index (p = 0.004), increased waist circumference (p = 0.004), increased waist-hip ratio (p < 0.001), and increased LDL cholesterol (p = 0.029), while the urban participants had a significantly higher prevalence of insufficient physical activity (p < 0.001). CONCLUSIONS: In the examined sample, the prevalence of cardiovascular risk factors is generally high. Participants from rural areas are significantly more susceptible to cardiovascular risk factors than participants from urban areas.

The Influence of the COVID-19 Pandemic on Hospitalizations for Ambulatory Care-Sensitive Conditions in Split-Dalmatia County, Croatia.

Background and Objectives: We aimed to explore whether the COVID-19 pandemic influenced hospitalizations for ambulatory care-sensitive conditions (ACSCs) in Split-Dalmatia County, Croatia. Materials and Methods: We performed a cross-sectional comparative study using two different time periods, the pre-pandemic (from March 2019 to February 2020) and the pandemic period (from March 2020 to February 2021), to explore the possible influences that the COVID-19 pandemic had on hospitalizations for ACSCs. The ACSCs were classified into the categories of vaccine-preventable, chronic, and acute disease. The indicators were statistically analyzed. Results: During the pandemic, a decrease in the total number of hospitalizations and ACSC hospitalizations was recorded. The relative risk for having any ACSC hospitalization in the pandemic period compared to the pre-pandemic period was 0.67 (95% CI, 0.64-0.71; p = 0). The risk reduction was seen in all three categories of vaccine-preventable ACSCs, chronic disease, and acute disease. Large reductions were found in the relative risk of hospitalization for COPD and asthma. Considering the mode of discharge, there was a statistically significantly higher risk of ACSCs with fatal outcomes during the pandemic than in the pre-pandemic period (relative risk 1.31; 95% CI, 1.01-1.7; p = 0.0197). Conclusions: The results of this study show that the COVID-19 pandemic influenced the total number of hospitalizations as well as hospitalizations relating to ACSCs. Certainly, one of the reasons for these changes was due to organizational changes in the working of the entire health system due to the COVID-19 pandemic.

Detection of Macrolide and/or Fluoroquinolone Resistance Genes in Mycoplasma genitalium Strains Isolated from Men in the Northwest Region of Croatia in 2018-2023.

Mycoplasma genitalium (M. genitalium) poses a significant public health challenge due to its association with non-gonococcal urethritis (particularly in men) and antimicrobial resistance. However, despite the prevalence of M. genitalium infections and the rise in resistance rates, routine testing and surveillance remain limited. This is the first study from Croatia that aimed to assess the prevalence and trends of resistance in M. genitalium strains isolated from male individuals by detecting macrolide and fluoroquinolone resistance genes. The study also aimed to explore the factors associated with resistance and changes in resistance patterns over time. Urine samples collected from male individuals in the Zagreb County and northwest region of Croatia between 2018 and 2023 were tested for M. genitalium with the use of molecular methods. Positive samples were subjected to DNA extraction and multiplex tandem polymerase chain reaction (MT-PCR) targeting genetic mutations associated with macrolide (23S rRNA gene) and fluoroquinolone (parC gene) resistance. Of the 8073 urine samples tested from 6480 male individuals (and following the exclusion of repeated specimens), we found that the prevalence of M. genitalium infection was 2.2%. Macrolide resistance was observed in 60.4% of strains, while fluoroquinolone resistance was found in 19.2%. Co-resistance to both antibiotics was present in 18.2% of cases. A statistically significant increase in fluoroquinolone resistance was noted over the study period (p = 0.010), but this was not evident for azithromycin resistance (p = 0.165). There were no statistically significant differences in resistance patterns between age groups, whereas re-testing of patients revealed dynamic changes in resistance profiles over time. The high burden of macrolide resistance and increasing fluoroquinolone resistance underscore the urgent need for comprehensive resistance testing and surveillance programs. The implementation of resistance-guided treatment strategies, along with enhanced access to molecular diagnostics, is pivotal for effectively managing M. genitalium infections.

Monitoring HPV Prevalence and Risk Cofactors for Abnormal Cytology in the Post-Vaccination Period among Croatian Women.

The incidence and mortality rate of cervical cancer in Croatia remains a health challenge despite screening efforts. Besides the persistent infection with HPV, the development of cancer is also associated with some cofactors. The goal of this study was to assess circulating HPV genotypes and risk factors for the development of cervical precancer after almost 16 years from the onset of HPV vaccination in Croatia. In this study, a total of 321 women attending gynecological care were evaluated. Relevant medical and demographic information, including cytology, were collected. HPV genotyping was performed by PCR. Comparing the HPV types found in circulation in the pre-vaccination (1999-2015) and post-vaccination periods (2020-2023), a statistically significant reduction in HPV 31 was noted, while the overall prevalence increased in the post-vaccination period. Besides the expected HPV positivity as a risk factor, the history of smoking was associated with LSIL or worse cytology at enrollment. For the first time, this population study revealed a statistically significant shift in the HPV genotype in the post-vaccination period, as well as the confirmation of risk factors for the development of abnormal cytology among Croatian women.

The role of longevity-related genetic variant interactions as predictors of survival after 85 years of age.

Genome-wide association studies and candidate gene studies have identified several genetic variants that might play a role in achieving longevity. This study investigates interactions between pairs of those single nucleotide polymorphisms (SNPs) and their effect on survival above the age of 85 in a sample of 327 Croatian individuals. Although none of the SNPs individually showed a significant effect on survival in this sample, 14 of the 359 interactions tested (between SNPs not in LD) reached the level of nominal significance (p<0.05), showing a potential effect on late-life survival. Notably, SH2B3 rs3184504 interacted with different SNPs near TERC, TP53 rs1042522 with different SNPs located near the CDKN2B gene, and CDKN2B rs1333049 with different SNPs in FOXO3, as well as with LINC02227 rs2149954. The other interaction pairs with a possible effect on survival were FOXO3 rs2802292 and ERCC2 rs50871, IL6 rs1800795 and GHRHR rs2267723, LINC02227 rs2149954 and PARK7 rs225119, as well as PARK7 rs225119 and PTPN1 rs6067484. These interactions remained significant when tested together with a set of health-related variables that also had a significant effect on survival above 85 years. In conclusion, our results confirm the central role of genetic regulation of insulin signalling and cell cycle control in longevity.

Seroepidemiology of Herpes Simplex Viruses Type 1 and 2 in Pregnant Women in Croatia.

Background and Objectives: Herpes simplex viruses (HSV-1 and HSV-2) are one of the most widespread causes of human viral infections. In Croatia, only two published studies have analyzed the seroprevalence of HSV infections in childbearing-aged and pregnant women (2005-2010), while more recent data are lacking. This study aimed to analyze the prevalence and risk factors for HSV-1 and HSV-2 infections among pregnant women in Croatia in the period from 2011 to 2021. Materials and Methods: This study included 667 pregnant women aged 16-45 years submitted for HSV-1 and HSV-2 serology testing. Serum samples were initially screened for HSV-1 and HSV-2 IgM and IgG antibodies using a commercial ELISA test with a confirmation of HSV-2-positive samples using an immunoblot assay. Results: The overall IgG seroprevalence rates were 69.9% for HSV-1 and 3.8% for HSV-2. A significant gradual increase in the HSV-2 seroprevalence with age was observed from 0.5% in participants under 30 years to 8.3% in participants above 40 years. The HSV-1 seroprevalence was stable up to 40 years (70.0 and 68.3%, respectively), with an increase to 86.1%, but this difference did not reach statistical significance. Area of residence (urban or suburban/rural), geographic region (continental or coastal), and obstetric history (normal pregnancy or unfavorable obstetric history) were not associated with HSV-1 and HSV-2 seroprevalence. Older age was found to be a significant risk factor for HSV-2 seropositivity in both univariate and multivariate risk analysis. Conclusions: HSV-1 infection is widely prevalent among pregnant women with a stable trend over time. However, a declining trend in the HSV-2 seroprevalence was observed compared to 2005-2010. Serological screening in pregnant women is important in identifying seronegative women who are susceptible to HSV infection as well as seropositive women who are at risk for genital herpes recurrence during delivery.

Epidemiology of Bartonella henselae infection in pet and stray cats in Croatia with risk factors analysis.

BACKGROUND: Cats are the primary reservoirs of the bacterium Bartonella henselae, the main cause of cat-scratch disease in humans. The main vector of the bacterium is the cat flea, Ctenocephalides felis. In southeastern Europe, data are lacking on the prevalence of B. henselae infection in cats, the strains of B. henselae involved and the risk factors associated with the infection. METHODS: Blood samples collected in ethylenediaminetetraacetic acid-containing tubes from 189 domestic cats (156 pet cats and 33 stray cats) from Zagreb, the capital city of Croatia, and 10 counties throughout Croatia were cultured for Bartonella spp. Following culture, bacterial isolates were genotyped at eight loci after using PCR to amplify 16S ribosomal RNA (rRNA) and the internal transcribed spacer region between the 16S and 23S rRNA sequences. Univariate and multivariate logistic regression were used to identify risk factors for B. henselae infection in cats. RESULTS: Bartonella spp. was detected in 31 cats (16.4%), and subsequent genotyping at the eight loci revealed B. henselae in all cases. Thirty complete multilocus sequence typing profiles were obtained, and the strains were identified as four sequence types that had been previously reported, namely ST5 (56.7%), ST6 (23.3%), ST1 (13.3%) and ST24 (3.3%), as well as a novel sequence type, ST33 (3.3%). The univariate analysis revealed a significantly higher risk of B. henselae infection in cats residing in coastal areas of Croatia (odds ratio [OR] 2.592, 95% confidence interval [CI] 1.150-5.838; P = 0.0191) and in cats with intestinal parasites (OR 3.207, 95% CI 1.088-9.457; P = 0.0279); a significantly lower risk was identified in cats aged > 1 year (OR 0.356, 95% CI 0.161-0.787; P = 0.0247) and in cats sampled between April and September (OR 0.325, 95% CI 0.147-0.715; P = 0.005). The multivariate analysis that controlled for age showed a positive association with the presence of intestinal parasites (OR 4.241, 95% CI 1.243-14.470; P = 0.0119) and coastal residence (OR 2.567, 95% CI 1.114-5.915; P = 0.0216) implying increased risk of infection, and a negative association with sampling between April and September (OR 0.379, 95% CI 0.169-0.848; P = 0.018) implying a decreased risk of infection. After controlling for the season, an increased risk of infection remained for the coastal region (OR 2.725, 95% CI 1.200-6.186; P = 0.012). CONCLUSIONS: Bartonella henselae is prevalent throughout Croatia and is a public health threat. Environmental and host factors can significantly affect the risk of infection, and these should be explored in more detail. The presence of intestinal parasites highlights the need to eliminate the flea vector, Ctenocephalides felis, as the most effective approach to control infections in cats and humans.

Levels of depression, anxiety and subjective happiness among health sciences students in Croatia: a multi-centric cross-sectional study.

BACKGROUND: Previous studies have shown that symptoms of depression and anxiety were highly prevalent among health sciences students. This may lead to other professional and personal difficulties and a decrease in individuals' well-being. This study aimed to analyze levels of depression, anxiety and subjective happiness among health sciences students in Croatia. METHODS: We conducted a cross-sectional study in 10 higher education institutions in Croatia during March 2023. Eligible participants were health sciences students. Participants filled out an online survey consisting of sociodemographic questions and validated scales for determining the levels of depression (9-question Patient Health Questionnaire, PHQ-9), anxiety (General Anxiety Disorder 7-item scale, GAD-7), and happiness (Subjective Happiness Scale, SHS). RESULTS: Of 7460 invited students, 2137 students participated in the study (29% response rate). There were 41.4% of students that exhibited at least mild depressive symptoms, with 8% of students exhibiting moderately severe symptoms and 1.8% severe depressive symptoms. Mild anxiety was found in 36.8%, moderate anxiety in 23.9% and severe anxiety in 15.8% of students. The median SHS score was 19 (15.25-22). Women students had significantly higher levels of depression (p < 0.001) and anxiety (p < 0.001) than their men peers. Students in earlier study years showed higher levels of depression, anxiety and lower levels of subjective happiness compared to those in later study years. Students with lower self-assessed financial status had higher levels of depression (p < 0.001) and anxiety (p < 0.001). Students that failed an academic year had higher levels of depression (p < 0.001), but lower levels of anxiety (p = 0.005). CONCLUSION: In this study, we have shown that health sciences students exhibit high levels of depression and anxiety, at rates exceeding those in the general population reported in other studies. Our results may help educational institutions to put greater effort into the battle against mental health stigma, foster acceptance of mental health issues and encourage students to seek help when needed. Adequate mental health services are needed at universities to promote timely diagnosis and treatment of mental health problems.

Bloodstream Infections in Pediatric Oncology Patients: Bacterial Pathogen Distribution and Antimicrobial Susceptibility at the University Hospital Centre Zagreb, Croatia-A 5-Year Analysis.

The epidemiology of bacterial pathogens causing bloodstream infections (BSIs) in pediatric hematology/oncology patients is changing and resistance to antimicrobial agents is globally spread. We retrospectively assessed demographic, clinical, and microbiologic data of BSIs during a 5-year period at a pediatric hematology/oncology unit from January 1, 2017, to December 31, 2021, at the University Hospital Centre Zagreb, Zagreb, Croatia. In 66 pediatric patients with malignancies, 93 BSI episodes were registered and 97 bacterial isolates were cultured. The Gram-positive versus Gram-negative ratio was 67 (69.1%) versus 30 (30.9%). Coagulase-negative staphylococci (48; 49.6%) were the most frequent isolates, followed by Enterobacterales (17; 17.5%) and Staphylococcus aureus (6; 6.2%). Multidrug resistance isolates included extended spectrum ß-lactamase producers (n=3). Resistance rates to piperacillin/tazobactam, cefepime, and meropenem in Gram-negative isolates were 15.4%, 14.3%, and 0.0%, respectively. Gram-positive bacteria are the most common cause of BSI in our patients. Resistance rates to piperacillin/tazobactam and cefepime in Gram-negative isolates make meropenem a better choice for empirical antimicrobial treatment. As national and hospital data may differ, the surveillance of pathogen distribution and antimicrobial susceptibility in pediatric hematology/oncology wards is necessary to adjust empirical treatment accordingly.

Increased number of cryptosporidiosis cases with travel history to Croatia might be related to swimming pools, Germany, 2023.

In August and September 2023, an unusually high number of cryptosporidiosis cases identified by routine German surveillance had travelled to Croatia (n = 23). Nine cases had stayed in the same camping resort and seven further cases had stayed at other camping sites within 15 km. Based on our standardised questionnaires, the most likely source of infection was swimming pools (93%). Further environmental investigations on site might reveal potential common sources of contamination that could be targeted by control measures.

SPATIAL AND SOCIAL ASPECTS OF THE 1918-1919 PANDEMIC FLU IN SPLIT / Prostorna i socijalna obiljezja pandemijske gripe 1918. - 1919. u Splitu

The Spanish flu pandemic is considered the largest and most dangerous epidemic at the beginning of the 20th century affecting most of the world today. The Spanish flu pandemic did not bypass the territory of Croatia (at the time, Croatia was part of the Austro-Hungarian Monarchy), nor its southern coastal region of Dalmatia and the city of Split. Using the example of the city Split, the paper analyses the spatial and demographic determinants of population mortality from the Spanish flu between 1918 and 1919, i.e., from March 1918 to April 1919. The paper is based on the data of the Church Death registers kept in the Archbishop's Archives in Split and newspaper articles. The analysis of the spatial distribution of mortality within urban settlements showed that the number of deaths per urban settlement

Fragility spinal fractures among cirrhotic liver transplant candidates in Croatia.

INTRODUCTION: Existing data on fragility spinal fractures prevalence in liver transplant candidates are scarce and inconsistent. This may be due to other comorbidities, besides hepatic osteodystrophy (HO), that contribute to bone loss and fragility fracture prevalence in chronic liver disease (CLD). OBJECTIVES: The aim of this study was to investigate the prevalence of spinal thoracic and lumbar fragility fractures among cirrhotic, non-chronic kidney disease (CKD), non-diabetic liver transplant candidates and to explore their relationship with clinical characteristics, laboratory markers and dual-energy x-ray absorptiometry (DXA) results. MATERIAL AND METHODS: This cross-sectional observational study was conducted at Merkur University Hospital, Croatia, between February 2019 and May 2023. Adult patients with liver cirrhosis referred for liver transplantation were included. Patients with acute infection, CKD, diabetes mellitus, malignancies, inflammatory bone diseases and those on corticosteroid or antiresorptive therapy were excluded. Clinical, laboratory and radiological assessment was carried out and patients were accordingly allocated into non-fractured and fractured group for the purpose of statistical analysis. RESULTS: A total of 90 patients were included in the study. There was 123 fractures, 87 (70.7 %) in the thoracic and 36 (29.3 %) in the lumbar region. Eighty-nine (72.4 %) fractures were grade 1, 31 (25.2 %) were grade 2 and 3 (2.4 %) were grade 3. Patients in the fractured group were significantly older (p < 0.001). No significant differences between fractured and non-fractured group according to laboratory and DXA parameters were noted. Subgroup with lumbar fractures had significantly lower bone mineral density values at L1-L4 region. Statistically significant negative correlation between bone specific alkaline phosphatase (BALP) and hip total BMD (rho = -0.414, p < 0.001) and spine total BMD (rho = -0.258, p = 0.014) values was found. CONCLUSION: Present study confirmed detrimental impact of CLD and HO on bone strength. DXA measurement correlated with the presence of lumbar fragility fractures. A combination of standard X-ray imaging and DXA is needed for adequate bone evaluation in pretransplant period and BALP could be useful for detecting HO in CLD. Searching for other risk factors and implementing bone turnover markers and additional imaging techniques for bone loss evaluation in liver transplant candidates is needed.

Prevalence of occult hepatitis B virus infection and characterisation of hepatitis B surface antigen mutants among adults in western Croatia.

INTRODUCTION AND OBJECTIVES: Occult hepatitis B virus (HBV) infection (OBI) is characterised by low levels of hepatitis B virus (HBV) DNA in the blood/liver of patients with negative hepatitis B surface antigen (HBsAg). This study aimed to determine the OBI prevalence and virological characteristics (viral genotypes and HBsAg mutants) in patients with an "anti-HBc only" serological profile. MATERIALS AND METHODS: A total of 24 900 serum samples were routinely screened for hepatitis B markers over a five-year period. All anti-HBc-positive/HBsAg-negative/anti-HBs-negative sera were selected and analysed for the presence of HBV DNA. Mutational analyses of the HBs gene and polymerase gene sequences were performed. RESULTS: 1749 (7.02%) sera were anti-HBc positive, and 113 (0.45%) sera had an "anti-HBc only" serological profile (HBsAg/anti-HBs negative). HBV DNA was detected in 12/113 (10.61%) "anti-HBc only" positive sera, representing 0.048% of all routinely tested samples. Due to extremely low viremia, HBV genome was successfully sequenced in only two sera where subgenotype D3 was confirmed. Mutational analyses of the S gene revealed multiple missense mutations. In addition to the M133I, Y134F, and G145R mutations, already associated with diagnostic escape, we also found nine novel OBI-related S-gene mutations - S136Y, F158L, K160N, E164G, S167L, A168V, L175S, S210I and F212C. CONCLUSIONS: We detected multiple known and novel S gene mutations in 2/12 (16.6%) OBI cases, nevertheless, further studies are required to determine their role in the pathogenesis of OBI. Understanding the frequencies of clinically relevant HBV mutations may contribute to improvement of diagnostic protocols.

Cancer morbidity in Croatian cats: Retrospective study on spontaneously arising tumors (2009-2019).

One of the current leading causes of death in pet cats is neoplasia. The scarcity of data on tumor prevalence in cats from south-eastern Europe led to this retrospective study which reports an abundant collection of tumors diagnosed in cats living in Croatia. Archived histopathology findings (diagnosis, cell origin, and biological behavior), and epidemiological data (breed, sex, age, and anatomical site of the tumor) of the cats diagnosed with at least one tumor were collected and statistically analyzed. For the research, 2338 archived feline samples, obtained from 2009-2019, have been reviewed. In 659 samples (28.1%) one or more tumors were diagnosed. Tumors have most often affected non-pedigree cats, cats between 7 and 15 years of age, and female cats. Most tumors showed malignant behavior (85.7%). The most frequent diagnoses were mammary adenocarcinoma, lymphoma, and squamous cell carcinoma. Regarding origin, most samples were epithelial (46.9%), but mesenchymal (28.7%) and lymphoid tumors (17.0%) were also frequently found. Tumors most often occurred in the skin and subcutis, mammary gland (in females), digestive system, and hemolymphatic system (in males). Statistical analysis did not reveal breed, sex, and age predisposition, but female animals and non-pedigree cats were significantly more likely to develop malignant tumors. This detailed study gives insight into the most frequent tumors that occur in cats from Croatia, and the data reported here is likely applicable to other countries. This data will contribute to clinical veterinary oncology, as it presents the most extensive tumor index of cats in south-eastern Europe.

Characterization of Human Immunodeficiency Virus-1 Transmission Clusters and Transmitted Drug-Resistant Mutations in Croatia from 2019 to 2022.

Molecular epidemiology of HIV-1 infection is challenging due to the highly diverse HIV-genome. We investigated the genetic diversity and prevalence of transmitted drug resistance (TDR) followed by phylogenetic analysis in 270 HIV-1 infected, treatment-naïve individuals from Croatia in the period 2019-2022. The results of this research confirmed a high overall prevalence of TDR of 16.7%. Resistance to nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside RTIs (NNRTIs), and protease inhibitors (PIs) was found in 9.6%, 7.4%, and 1.5% of persons, respectively. No resistance to integrase strand-transfer inhibitors (INSTIs) was found. Phylogenetic analysis revealed that 173/229 sequences (75.5%) were part of transmission clusters, and the largest identified was T215S, consisting of 45 sequences. Forward transmission was confirmed in several clusters. We compared deep sequencing (DS) with Sanger sequencing (SS) on 60 randomly selected samples and identified additional surveillance drug resistance mutations (SDRMs) in 49 of them. Our data highlight the need for baseline resistance testing in treatment-naïve persons. Although no major INSTIs were found, monitoring of SDRMs to INSTIs should be continued due to the extensive use of first- and second-generation INSTIs.