The Effects of a Pulmonary Rehabilitation Programme on Functional Capacity and Strength of Respiratory Muscles in Patients with Post-COVID Syndrome.

Aim: The aim of this study was to estimate the effects of a pulmonary rehabilitation programme (PR) on the functional capacity and respiratory muscle strength of patients with post-COVID syndrome. Methods: A cross-sectional study was conducted using hospital data on patients who participated in a pulmonary rehabilitation programme at the Clinic for Lung Diseases, University Hospital Centre Zagreb, Croatia, between January 2021 and December 2022. Data on the spirometry, respiratory muscle strength, and functional exercise capacity of patients were collected at baseline and three weeks after the start of rehabilitation. The study included 80 patients (43 females, 37 males) with a mean age of 51±10 years. Results: A significant increase in respiratory muscle strength (P<0.001) was observed after pulmonary rehabilitation, with effect sizes ranging from small to large (Cohen's d from 0.39 to 1.07), whereas the effect for PImax expressed as a percentage was large (Cohen's d=0.99). In addition, the pulmonary rehabilitation programme significantly improved the parameters of the six-minute walk test in patients, and the parameters of lung function, FVC, FEV1, and DLCO also improved significantly after PR (P<0.05). Conclusion: The results showed that the pulmonary rehabilitation programme has clinically significant effects on functional capacity and respiratory muscle strength in patients with post-COVID syndrome.

Myocardial Infarction Susceptibility and the MTNR1B Polymorphisms.

Melatonin is a circadian hormone with antioxidant properties that protects against myocardial ischemia-reperfusion injury. Genetic variations of the melatonin receptor 1B gene (MTNR1B) play an important role in the development of type 2 diabetes, a risk factor for cardiovascular diseases. Accordingly, MTNR1B polymorphisms are crucial in numerous disorders of the cardiovascular system. Therefore, the aim of the present study was to investigate a possible association of MTNR1B polymorphisms with chronotype and susceptibility to myocardial infarction. The present case-control study included 199 patients with myocardial infarction (MI) (57% men) and 198 control participants (52% men) without previous cardiovascular diseases who underwent genotyping for the MTNR1B polymorphisms rs10830963, rs1387153, and rs4753426 from peripheral blood samples. Chronotype was determined using the Morningness-Eveningness Questionnaire (MEQ). As estimated by the chi-square test, no significant association was found in the distribution of alleles and genotypes between myocardial infarction patients and controls. In addition, there was no association between MTNR1B polymorphisms and chronotype in MI patients. As some previous studies have shown, the present negative results do not exclude the role of the MTNR1B polymorphisms studied in the development of myocardial infarction. Rather, they may indicate that MTNR1B polymorphisms are a minor risk factor for myocardial infarction.

Geographical and Temporal Distribution of Radiologists, Computed Tomography and Magnetic Resonance Scanners in Croatia.

The aim of the study was to analyse the temporal and geographic distribution of radiologists, computed tomography and magnetic resonance scanners in Croatia. In this observational study we estimated radiologists' number per 100,000 population for 1997, 2006, and 2017 and compared private and public CT and MR scanners between 2011 and 2018. We analyzed the availability of radiologists and scanners, and the relationship between the radiological workforce and economic strength among counties. The workforce increased significantly from 1997 to 2017 and was associated with economic strength categories in 2017. In 2018, there were more CT scanners in the public sector, while MR scanners were distributed evenly. In 2011, there was similar distribution of CT and MR between sectors, while in 2018 there were significantly more public CT scanners. Counties with a medical school had significantly more radiologists and MR scanners. The high-to-low ratios per CT and MR were 11 and 8.2, suggesting inequality of health care. Croatia significantly increased its radiological workforce; however, cross-county inequality remained. Counties with higher economic strength and medical schools have better availability of radiologists and equipment. To ensure the sustainable activity of the health care system, a precise estimate of supply and demand of radiology services is needed.

The analysis of waiting time and utilization of computed tomography and magnetic resonance imaging in Croatia: a nationwide survey.

AIM: To assess the variation in the waiting time for diagnostic imaging (DI) services among Croatian public hospitals and the utilization of computed tomography (CT) and magnetic resonance imaging (MRI) scanners. METHODS: We analyzed aggregated data from public hospitals. Counties were classified according to economic strength, and utilization was expressed as the average number of exams per machine. We compared the waiting times for 2018 and utilization for 2015 according to hospital category (high and low level) and economic strength by county. RESULTS: The waiting time was longer for MRI compared with CT, 268 vs 77.61 days. Overall CT waiting time was in the unfavorable European Health Consumer Index category. High-level hospitals had longer waiting time for MRI and CT. The waiting time positively correlated with economic strength for MRI (P=0.019), but not for CT. In low-level hospitals, MRI utilization ranged from 104 to 6032, whereas CT utilization ranged from 48 to 17852. In high-level hospitals, MRI utilization ranged from 3846 to 11 026, while CT utilization ranged from 503 to 17 234. CT (P=0.041) and MRI (P=0.031) utilization in high-level hospitals was significantly higher than in low-level hospitals. CONCLUSION: The waiting times for CT and MRI were exceptionally long regardless of the hospital category, with highly varying utilization. Croatia performed more exams per scanner compared with other EU countries, but not significantly so. High-level hospitals' utilization was significantly higher than that of low-level hospitals, and CT utilization was significantly higher than EU average, while the difference for MRI utilization was not significant.

Epidemiological and clinical features of primary biliary cholangitis in two Croatian regions: a retrospective study.

AIM: To assess the measures of disease frequency and determine the clinical features of primary biliary cholangitis (PBC) in two Croatian regions. METHODS: Databases of two tertiary hospitals, one located in the continental and one in the coastal region of Croatia, were retrospectively searched for PBC patients diagnosed from 2007 to 2018. Epidemiologic data analysis was restricted to patients from each hospital's catchment area. We analyzed factors related to response to therapy and event-free survival (EFS), defined as absence of ascites, variceal bleeding, encephalopathy, hepatocellular carcinoma, liver transplantation (LT), or death. In addition, we determined clinical and demographic data of transplanted PBC patients. RESULTS: Out of 83 PBC patients, 86.7% were female, with a median age at diagnosis of 55 years. Average PBC incidence for the 11-year period was 0.79 and 0.89 per 100000 population, whereas the point prevalence on December 31, 2017 was 11.5 and 12.5 in the continental and coastal region, respectively. Of 76 patients with complete medical records, 21% had an advanced disease stage, 31.6% had an associated autoimmune condition, and all received ursodeoxycholic acid. EFS rate at 5 years was 95.8%. In an age and sex-adjusted multivariate Cox regression model, the only factor significantly associated with inferior EFS was no response to therapy (HR=18.4; P=0.018). Of all Croatian patients who underwent LT, 3.8% had PBC, with the survival rate at 5 years after LT of 93.4%. CONCLUSION: This study gives pioneer insights into the epidemiological and clinical data on PBC in Croatia, thus complementing the PBC map of Southeast Europe.

Quantification of coronary atherosclerotic burden with coronary computed tomography angiography: adapted Leaman score in Croatian patients.

The aim of the study was to quantify the total coronary atherosclerotic burden in patients with suspected coronary artery disease (CAD) defined by coronary computed tomography adapted Leaman score (CT-LeSc) and to estimate its cut-off level for high coronary atherosclerotic burden. We enrolled 434 consecutive patients referred to coronary computed tomography angiography, of which 261 patients fulfilled the study inclusion criteria. Demographic and clinical characteristics, as well as CAD risk factors were obtained. CAD pre-test probabilities were estimated by the Diamond-Forrester model and Morise score. The coronary atherosclerotic burden was estimated using CT-LeSc. As a cut-off for a high coronary atherosclerotic burden, we used 3rd tercile (Tc3) (CT-LeSc ≥ 5.52). We evaluated the association of clinical characteristics and risk factors with Tc3 in univariate and multivariate analysis. There were 60.9% males and 39.1% females, 81% of patients had above-normal weight, 68.2% hypertension, 54.0% dyslipidemia, 15.3% diabetes mellitus, 12.3% positive smoking history and 11.9% had a family history of CAD. According to the Diamond-Forrester model and Morise score the majority of patients had intermediate risk, 59.7 and 52.8%, followed by the high-risk group, 36.0 and 34.4%, respectively. Age, dyslipidemia, hypertension and pre-test risk scores in the univariate analysis significantly predicted Tc3. In the multivariate analysis, male sex (p = 0.004), dyslipidemia (p = 0.002) and coronary calcium score (< 0.001) were identified as predictors of Tc3. CT-LeSc quantified the total coronary atherosclerotic burden and showed an association of risk factors and pre-test probabilities with Tc3.

Individual multi-locus heterozygosity is associated with lower morning plasma cortisol concentrations.

OBJECTIVE: Stress is implicated as a risk factor for numerous illnesses in humans, putatively in part mediated by biological responses to stress, such as elevated cortisol concentrations. The theory of genetic homoeostasis suggests that individual heterozygosity facilitates compensation for environmental stresses. We hypothesized that heterozygosity ameliorates the biological response to a given level of perceived stress, reflected in lower plasma cortisol concentrations. DESIGN: We examined the role of heterozygosity in the association between perceived psychological stress and morning cortisol concentrations in 854 individuals from the isolated island of Vis, Croatia. METHODS: Cortisol concentrations were measured in morning plasma samples. A total of 1184 autosomal microsatellite markers were genotyped and individual multi-locus heterozygosity (MLH) was calculated as the proportion of heterozygous markers. The General Health Questionnaire with 30 items (GHQ-30) was used to assess the degree of psychological distress. RESULTS: MEAN MLH WAS 34.850.45% (RANGE: 31.97-36.22%). Psychological distress (GHQ Likert score >31) was more prevalent in women (37 vs 18% in men, P<0.0001), in less educated people (ß=-0.35 per year in school, P<0.001) and in lower socio-economic classes (ß=-3.59, P<0.0001). Cortisol concentrations were positively associated with psychological distress (ß=2.20, P=0.01). In a regression model adjusted for age, BMI, education and GHQ-30 score, MLH was independently and inversely associated with morning plasma cortisol concentrations (P=0.005). CONCLUSION: More heterozygous individuals, as measured by microsatellite markers, had lower morning plasma cortisol concentrations for a given level of perceived psychological stress. This may be important, as higher cortisol concentrations may increase the allostatic load and be associated with a higher risk of stress-related illness.

Setting research priorities to reduce global mortality from preterm birth and low birth weight by 2015.

AIM: This paper aims to identify health research priorities that could improve the rate of progress in reducing global neonatal mortality from preterm birth and low birth weight (PB/LBW), as set out in the UN's Millennium Development Goal 4. METHODS: We applied the Child Health and Nutrition Research Initiative (CHNRI) methodology for setting priorities in health research investments. In the process coordinated by the World Health Organization in 2007-2008, 21 researchers with interest in child, maternal and newborn health suggested 82 research ideas that spanned across the broad spectrum of epidemiological research, health policy and systems research, improvement of existing interventions and development of new interventions. The 82 research questions were then assessed for answerability, effectiveness, deliverability, maximum potential for mortality reduction and the effect on equity using the CHNRI method. RESULTS: The top 10 identified research priorities were dominated by health systems and policy research questions (eg, identification of LBW infants born at home within 24-48 hours of birth for additional care; approaches to improve quality of care of LBW infants in health facilities; identification of barriers to optimal home care practices including care seeking; and approaches to increase the use of antenatal corticosteriods in preterm labor and to improve access to hospital care for LBW infants). These were followed by priorities for improvement of the existing interventions (eg, early initiation of breastfeeding, including feeding mode and techniques for those unable to suckle directly from the breast; improved cord care, such as chlorhexidine application; and alternative methods to Kangaroo Mother Care (KMC) to keep LBW infants warm in community settings). The highest-ranked epidemiological question suggested improving criteria for identifying LBW infants who need to be cared for in a hospital. Among the new interventions, the greatest support was shown for the development of new simple and effective interventions for providing thermal care to LBW infants, if KMC is not acceptable to the mother. CONCLUSION: The context for this exercise was set within the MDG4, requiring an urgent and rapid progress in mortality reduction from low birth weight, rather than identifying long-term strategic solutions of the greatest potential. In a short-term context, the health policy and systems research to improve access and coverage by the existing interventions, coupled with further research to improve effectiveness, deliverability and acceptance of existing interventions, and epidemiological research to address the key gaps in knowledge, were all highlighted as research priorities.

The analysis of tobacco consumption in Croatia--are we successfully facing the epidemic?

Tobacco is the largest cause of morbidity and mortality. The aim of this study is to analyse several health and economically related indicators of tobacco consumption: smoking prevalence, standardized death rates (SDRs) from lung cancer and the proportion of GDP spent on tobacco in Croatia and other transitional countries--the Czech Republic, Slovakia, Poland, Hungary, Slovenia, Romania, and Bulgaria. The overall smoking prevalence in Croatia decreased by 5.2% during 1994-2005, more among females (-9.9%) than males (-0.3%). There is no significant difference in the smoking prevalence between Croatia (27.4%) and other countries. However, 33.8% of Croatian males smoked during 2002-2005, more than in Romania and the Czech Republic, and less than in Hungary and Poland. The prevalence of female smoking (21.7%) in Croatia is similar to the female smoking prevalence in Poland, the Czech Republic, and Hungary, but male smoking is predominant in all countries. The proportion of smokers among youth is above 20% and it is the highest in the Czech Republic (29.7%), followed by Hungary (26.7%), Slovenia (24.9%), Croatia (24.1%), and Poland (21.5%). The proportion of smokers among girls is higher than among boys in Slovenia, Hungary, the Czech Republic, and Croatia, contrary to Slovakia, Bulgaria, and Poland where boys smoke slightly more. There is no significant difference between the prevalence of smoking among girls in Croatia and Bulgaria, Poland, the Czech Republic, Hungary, Slovenia, and Slovakia. According to the SDR from lung cancer in males (70.3/100,000), Croatia is ranked high assuming the 3rd place, after Hungary (99.7) and Poland (72.0). With a SDR of 15.9/100,000 for females, Croatia is ranked slightly better--5th place. Tobacco consumption continues to be a major public health problem in transitional countries. Croatia conducted several campaigns and programmes in the past. However, results reveal that current anti-tobacco strategies are ineffective in reducing the smoking prevalence among men and youth. Men do not smoke less than a decade ago and, despite the observed decline among women, increasing trends are observed among teenage girls. Croatia should apply a comprehensive approach that would include raising awareness of health risks, restriction of smoking in public places, higher taxing, implementing stricter bans on advertising and promotion of tobacco as well as supporting smoking cessation. This last measure is believed to bring about some results in the medium term in targeted population groups, provided that it is supported by all health professionals. Otherwise, we may expect progress at the population level in the field of social stigmatization of smoking and wider intolerance to second-hand smoke. The full impact of smoking on the population health is yet to be seen and in the future it will undoubtedly remain one of the major contributors to the poor public health situation in Croatia.

Genome-wide association study identifies novel loci associated with circulating phospho- and sphingolipid concentrations.

Phospho- and sphingolipids are crucial cellular and intracellular compounds. These lipids are required for active transport, a number of enzymatic processes, membrane formation, and cell signalling. Disruption of their metabolism leads to several diseases, with diverse neurological, psychiatric, and metabolic consequences. A large number of phospholipid and sphingolipid species can be detected and measured in human plasma. We conducted a meta-analysis of five European family-based genome-wide association studies (N = 4034) on plasma levels of 24 sphingomyelins (SPM), 9 ceramides (CER), 57 phosphatidylcholines (PC), 20 lysophosphatidylcholines (LPC), 27 phosphatidylethanolamines (PE), and 16 PE-based plasmalogens (PLPE), as well as their proportions in each major class. This effort yielded 25 genome-wide significant loci for phospholipids (smallest P-value = 9.88×10(-204)) and 10 loci for sphingolipids (smallest P-value = 3.10×10(-57)). After a correction for multiple comparisons (P-value<2.2×10(-9)), we observed four novel loci significantly associated with phospholipids (PAQR9, AGPAT1, PKD2L1, PDXDC1) and two with sphingolipids (PLD2 and APOE) explaining up to 3.1% of the variance. Further analysis of the top findings with respect to within class molar proportions uncovered three additional loci for phospholipids (PNLIPRP2, PCDH20, and ABDH3) suggesting their involvement in either fatty acid elongation/saturation processes or fatty acid specific turnover mechanisms. Among those, 14 loci (KCNH7, AGPAT1, PNLIPRP2, SYT9, FADS1-2-3, DLG2, APOA1, ELOVL2, CDK17, LIPC, PDXDC1, PLD2, LASS4, and APOE) mapped into the glycerophospholipid and 12 loci (ILKAP, ITGA9, AGPAT1, FADS1-2-3, APOA1, PCDH20, LIPC, PDXDC1, SGPP1, APOE, LASS4, and PLD2) to the sphingolipid pathways. In large meta-analyses, associations between FADS1-2-3 and carotid intima media thickness, AGPAT1 and type 2 diabetes, and APOA1 and coronary artery disease were observed. In conclusion, our study identified nine novel phospho- and sphingolipid loci, substantially increasing our knowledge of the genetic basis for these traits.

Role of cell adhesion molecules in acute ischemic stroke.

BACKGROUND AND OBJECTIVES: The expression of soluble adhesion molecules inter-cellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1), besides activation of endothelial cells and transendothelial migration of leukocytes, play an important role in inflammation and progression of ischemic injury after acute stroke. The aim of this study was to determine serum levels of soluble ICAM-1 and VCAM-1 in patients with acute ischemic stroke and controls and correlate them according to etiological subtypes (thromboembolic or lacunar stroke), stroke severity and disability after acute stroke. PATIENTS AND METHODS: Hospital-based prospective study of acute stroke patients hospitalized between December 2008 and September 2009 at the University Hospital Sestre Milosrdnice in Zagreb, Croatia. METHODS: We enrolled 110 patients with acute ischemic stroke and 93 healthy individuals as controls. Serum concentrations of VCAM-1 and ICAM-1 were determined by means of quantitative sandwich enzyme immunoassay. Patients were classified according to etiological subtype, clinical severity of stroke and disability after stroke. RESULTS: There was no significant difference between levels of soluble adhesion molecules VCAM-1 and ICAM-1 in patients and in controls. Levels of VCAM-1 were significantly higher in patients with thromboembolic stroke than in controls. There was no significant correlation between levels of soluble adhesion molecules VCAM-1 and ICAM-1 and stroke severity and disability. There was marked biological interindividual variability in all patient groups. CONCLUSION: This study confirms the role of adhesion molecule VCAM-1 in the pathogenesis of acute thromboembolic stroke.

Genetic determinants of circulating sphingolipid concentrations in European populations.

Sphingolipids have essential roles as structural components of cell membranes and in cell signalling, and disruption of their metabolism causes several diseases, with diverse neurological, psychiatric, and metabolic consequences. Increasingly, variants within a few of the genes that encode enzymes involved in sphingolipid metabolism are being associated with complex disease phenotypes. Direct experimental evidence supports a role of specific sphingolipid species in several common complex chronic disease processes including atherosclerotic plaque formation, myocardial infarction (MI), cardiomyopathy, pancreatic beta-cell failure, insulin resistance, and type 2 diabetes mellitus. Therefore, sphingolipids represent novel and important intermediate phenotypes for genetic analysis, yet little is known about the major genetic variants that influence their circulating levels in the general population. We performed a genome-wide association study (GWAS) between 318,237 single-nucleotide polymorphisms (SNPs) and levels of circulating sphingomyelin (SM), dihydrosphingomyelin (Dih-SM), ceramide (Cer), and glucosylceramide (GluCer) single lipid species (33 traits); and 43 matched metabolite ratios measured in 4,400 subjects from five diverse European populations. Associated variants (32) in five genomic regions were identified with genome-wide significant corrected p-values ranging down to 9.08x10(-66). The strongest associations were observed in or near 7 genes functionally involved in ceramide biosynthesis and trafficking: SPTLC3, LASS4, SGPP1, ATP10D, and FADS1-3. Variants in 3 loci (ATP10D, FADS3, and SPTLC3) associate with MI in a series of three German MI studies. An additional 70 variants across 23 candidate genes involved in sphingolipid-metabolizing pathways also demonstrate association (p = 10(-4) or less). Circulating concentrations of several key components in sphingolipid metabolism are thus under strong genetic control, and variants in these loci can be tested for a role in the development of common cardiovascular, metabolic, neurological, and psychiatric diseases.

Prevalence of increased body weight and hypertension in the population of Croatian mainland and Adriatic Islands--are islanders really healthier?

The aim of this study was to compare the prevalence of the hypertension and the increased body weight (BMI) between Croatian mainland and Adriatic island population. The data from the Croatian Adult Health Survey (N = 9,070) served as an estimate for the mainland Croatian population, while the data from "1001 Dalmatian study" (N = 1,001) were collected from four Adriatic islands; Rab, Vis, Lastovo and Mljet. The prevalence of increased body weight and hypertension was calculated for the four age groups and analyzed using chi-square test. The results indicate that men from the islands less frequently had normal body mass index (P < 0.001), and were more frequently overweight (P < 0.001). The prevalence of overweight and obesity were similar between the island and mainland women. The percent of normotensive respondents in men was significantly lower in islands (P < 0.001), while the prevalence of newly diagnosed hypertension was significantly higher among islanders in both genders (P < 0.001). Despite the traditionally prevalent Mediterranean diet and overall more favorable lifestyle islanders may not be as healthy as previously studies suggested, in terms of cardiovascular risk factors prevalence. This might be related to the poor access to health care and preventive measures or low interest for health care especially among men on the islands, reflected in the higher prevalence of newly diagnosed hypertension. These findings suggest that island populations represent good candidates for disease awareness programs and health promotion interventions.

Genome-wide linkage analysis of serum creatinine in three isolated European populations.

There is increasing evidence for a role of genetic predisposition in the etiology of kidney disease, but linkage scans have been poorly replicated. Here we performed a genome-wide linkage analysis of serum creatinine on 2859 individuals from isolated villages in South Tyrol (Italy), Rucphen (The Netherlands) and Vis Island (Croatia), populations that have been stable and permanently resident in their region. Linkage of serum creatinine levels to loci on chromosomes 7p14, 9p21, 11p15, 15q15-21, 16p13, and 18p11 was successfully replicated in at least one discovery population or in the pooled analysis. A novel locus was found on chromosome 10p11. Linkage to chromosome 22q13, independent of diabetes and hypertension, was detected over a region containing the non-muscle myosin heavy chain type II isoform A (MYH9) gene (LOD score=3.52). In non-diabetic individuals, serum creatinine was associated with this gene in two of the three populations and in meta-analysis (SNP rs11089788, P-value=0.0089). In populations sharing a homogeneous environment and genetic background, heritability of serum creatinine was higher than in outbred populations, with consequent detection of a larger number of loci than reported before. Our finding of a replicated association of serum creatinine with the MYH9 gene, recently linked to pathological renal conditions in African Americans, suggests that this gene may also influence kidney function in healthy Europeans.

Genome-wide association study of anthropometric traits in Korcula Island, Croatia.

AIM: To identify genetic variants underlying six anthropometric traits: body height, body weight, body mass index, brachial circumference, waist circumference, and hip circumference, using a genome-wide association study. METHODS: The study was carried out in the isolated population of the island of Korcula, Croatia, with 898 adult examinees who participated in the larger DNA-based genetic epidemiological study in 2007. Anthropometric measurements followed standard internationally accepted procedures. Examinees were genotyped using HumanHap 370CNV chip by Illumina, with a genome-wide scan containing 316730 single nucleotide polymorphisms (SNP). RESULTS: A total of 11 SNPs were associated with the investigated traits at the level of P<10(-5), with one SNP (rs7792939 in gene zinc finger protein 498, ZNF498) associated with body weight, hip circumference, and brachial circumference (P=3.59-5.73 x 10(-6)), and another one (rs157350 in gene delta-sarcoglycan, SGCD) with both brachial and hip circumference (P=3.70-6.08 x 10(-6). Variants in CRIM1, a gene regulating delivery of bone morphogenetic proteins to the cell surface, and ITGA1, involved in the regulation of mesenchymal stem cell proliferation and cartilage production, were also associated with brachial circumference (P=7.82 and 9.68 x 10(-6), respectively) and represent interesting functional candidates. Other associations involved those between genes SEZ6L2 and MAX and waist circumference, XTP6 and brachial circumference, and AMPA1/GRIA1 and height. CONCLUSION: Although the study was underpowered for the reported associations to reach formal threshold of genome-wide significance under the assumption of independent multiple testing, the consistency of association between the 2 variants and a set of anthropometric traits makes CRIM1 and ITGA1 highly interesting for further replication and functional follow-up. Increased linkage disequilibrium between the used markers in an isolated population makes the formal significance threshold overly stringent, and changed allele frequencies in isolate population may contribute to identifying variants that would not be easily identified in large outbred populations.

Historic, demographic, and genetic evidence for increased population frequencies of CCR5Delta32 mutation in Croatian Island isolates after lethal 15th century epidemics.

AIM: To assess the frequency of 32 base pair deletion in CCR5 (CCR5Delta32), which has been shown to confer resistance to HIV infection in a homozygous form, in 10 isolated island communities of Dalmatia, Croatia, with different histories of exposure to epidemics during and since the medieval period. METHODS: In 2002, DNA analysis of 100 randomly selected individuals from each of the 10 isolated communities of 5 Croatian islands (Susak, Rab, Vis, Lastovo, and Mljet) showed high levels of 3-generational endogamy, indicating limited gene flow. Five of the communities were decimated by epidemics of unknown cause between 1449-1456, while the other 5 villages remained unaffected. Genotyping of the CCR5 gene was performed using the polymerase chain reaction method with primers flanking the region containing 32-bp deletion. RESULTS: The frequency of CCR5Delta32 in the 5 villages affected by the epidemic was 6.1-10.0%, and 1.0-3.8% in the 5 unaffected villages. The Delta32 mutation was found in 71 of 916 alleles among the individuals from the affected villages (7.5%), and in 24 of 968 alleles in unaffected villages (2.5%, chi(2)=27.3, P<10-6). A previous study in 303 random Croatian blood donors showed the frequency of the CCR5 Delta32 of 7.1% in the general population. The difference remained significant after correcting for population structure using both STRAT and STRUCTURE software and the genomic control test, to ensure results do not arise from the background genetic differences. CONCLUSION: Our results and historical evidence, suggest that the mid-15th century epidemic could have acted as a selection pressure for the CCR5Delta32 mutation.

The evaluation of the stroke unit in Croatia at the University Hospital Sestre milosrdnice, Zagreb: 1995-2006 experience.

This study evaluate the effects of the Stroke Unit (SU) in Croatia by comparing the in-hospital case fatality rate in the period before (1995-2000) and after (2001-2006) the implementation of SU and to compare the prevalence of risk factors, such as hypertension, diabetes mellitus (DM), atrial fibrillation (AF) and ischemic heart disease (IHD) among the patients who died. The study was conducted in twelve-year period during which 10,901 stroke patients were admitted to hospital and 1818 of them died. The endpoints were in-hospital case fatality rate and prevalence of risk factors among the patients who died. Before the SU period the case fatality rate was 20.1%, whereas afterwards it decreased significantly to 12.8% (p < 0.001). The relative risk (RR) was 1.57, while the estimate of the odds ratio (OR) showed a 71% increase in chances of death in the pre-SU period. The prevalence of DM, IHD and AF increased significantly, while hypertension was the only risk factor which significantly decreased (p < 0.001). The results showed that the implementation of SU care is associated with a significant reduction of in-hospital case fatality rate of acute stroke patients which strongly suggests that development of the SU network in Croatia should be given priority in the health management.

Epidemiology and etiology of childhood pneumonia.

Childhood pneumonia is the leading single cause of mortality in children aged less than 5 years. The incidence in this age group is estimated to be 0.29 episodes per child-year in developing and 0.05 episodes per child-year in developed countries. This translates into about 156 million new episodes each year worldwide, of which 151 million episodes are in the developing world. Most cases occur in India (43 million), China (21 million) and Pakistan (10 million), with additional high numbers in Bangladesh, Indonesia and Nigeria (6 million each). Of all community cases, 7-13% are severe enough to be life-threatening and require hospitalization. Substantial evidence revealed that the leading risk factors contributing to pneumonia incidence are lack of exclusive breastfeeding, undernutrition, indoor air pollution, low birth weight, crowding and lack of measles immunization. Pneumonia is responsible for about 19% of all deaths in children aged less than 5 years, of which more than 70% take place in sub-Saharan Africa and south-east Asia. Although based on limited available evidence, recent studies have identified Streptococcus pneumoniae, Haemophilus influenzae and respiratory syncytial virus as the main pathogens associated with childhood pneumonia.