Anthropological Analysis of Neolithic and Early Bronze Age Skeletons – A Classical and Molecular Approach (East Slavonia, Croatia)

Theories about the first Indo-European migration are numerous. Significant contribution in attempt to resolve these theories is given by analysing skeletal material from two biggest prehistoric archaeological sites from N-E Croatia. Eight skeletons of Star~evo culture from sites »Nama« and »Hotel« at Vinkovci (6100–5500 BC) and seven skeletons of Vu~edol culture from the site Vineyard Streim at Vu~edol near Vukovar (3000–2500 BC) were analysed. Methods of classical anthropological analysis tried to distinguish the differences among members of both populations, while the methods of molecular genetics were used in defining possible genetic structure of both ancient populations. Established differences speak on the behalf of the theory of Maria Gimbutas about the first Indo-European migration with a cattle breeding population from the east around 3500 BC

Laboratory testing of thrombophilia in children

Pojam trombofilija obično se koristi za opisivanje nasljednih i/ili stečenih stanja povezanih s povećanom sklonošću za nastanak tromboze i klinički se odnosi na pacijente koji razvijaju spontanu vensku tromboemboliju (VTE). VTE u dječjoj dobi jest multifaktorijalna bolest, a u većine djece prisutan je jedan ili više kliničkih čimbenika rizika. Osim stečenih čimbenika, kao što su primjena centralnoga venskog katetera, maligne bolesti, teške infekcije, nefrotski sindrom i nasljedni trombofilni poremećaji mogu pridonijeti razvoju VTE-a u novorođenčadi. Nasljedna trombofilija uključuje nedostatak antitrombina (AT), proteina C (PC), proteina S (PS) te mutacije gena za faktor V (FV G1691A, Leiden) i protrombin (FII G20210A). Druge trombofilije, koje su slabije karakterizirane i nisu nužno genetički uvjetovane, uključuju povišenu razinu homocisteina, lipoproteina(a), disfibrinogenemije, antifosfolipidni sindrom i povećane razine faktora VIII, IX i XI. Sukladno najnovijim smjernicama, novorođenčad i djeca s purpurom fulminans trebaju biti odmah testirana na nedostatak PC-a i PS-a, dok se probir na trombofiliju u djece s moždanim udarom ne preporučuje rutinski. U djece s višestrukim neobjašnjivim trombozama, posebice ako ukazuju na katastrofični antifosfolipidni sindrom, treba razmotriti testiranje na antifosfolipidna protutijela i nasljednu trombofiliju. Skupine pedijatrijskih pacijenata koje će najvjerojatnije imati koristi od testiranja na trombofiliju jesu adolescenti sa spontanim nastankom VTE-a, djeca/adolescenti s pozitivnom obiteljskom anamnezom te tinejdžerice s poznatom pozitivnom obiteljskom anamnezom koje planiraju uzimanje oralnih kontraceptiva koji sadrže estrogen. Manje je korisno testiranje provesti u novorođenčadi i djece u kojih je nastanak VTE-a povezan s primjenom venskog katetera. Zaključno, može se reći da testiranje na nasljednu trombofiliju treba provesti individualiziranim pristupom i samo u slučaju kada bi dobiveni rezultati mogli utjecati na poboljšanje ili promjenu tijeka liječenja.The term thrombophilia is commonly used to describe inherited and/or acquired conditions associated with an increased propensity for thrombosis and clinically refers to patients who develop spontaneous venous thromboembolism (VTE). VTE in childhood is a multifactorial disease, and one or more clinical risk factors are present in most children. In addition to acquired factors, such as the use of a central venous catheter, malignancies, severe infections and nephrotic syndrome and hereditary thrombophilic disorders may contribute to the development of VTE in newborns. Hereditary thrombophilia includes deficiency of antithrombin (AT), protein C (PC), protein S (PS) and mutations in the genes for factor V (FV G1691A, Leiden) and prothrombin (FII G20210A). Other thrombophilias, which are less well characterized and not necessarily genetically determined, include elevated levels of homocysteine, lipoprotein(a), dysfibrinogenemia, antiphospholipid syndrome, and increased levels of factors VIII, IX, and XI. According to the latest guidelines, newborns and children with purpura fulminans should be tested immediately for PC and PS deficiency, while screening for thrombophilia in children with stroke is not routinely recommended. In children with multiple unexplained thromboses, especially if they indicate catastrophic antiphospholipid syndrome, testing for antiphospholipid antibodies and hereditary thrombophilia should be considered. The pediatric patient groups most likely to benefit from thrombophilia testing are adolescents with spontaneous VTE, children/adolescents with a positive family history, teenage girls with a known positive family history who plan to take estrogen containing oral contraceptives. It is less useful to perform testing in newborns and children in whom the occurrence of VTE is associated with the use of a venous catheter. In conclusion, it can be said that testing for hereditary thrombophilia should be carried out with an individualized approach and only if the obtained results could affect the improvement or change of the course of treatment

Zajednički poslijediplomski doktorski studij zdravstvenih znanosti

Ovaj doktorski studij omogućio bi studentima potrebnu razinu znanja, vještina i stavova temeljenih na interdisciplinarnom pristupu u poimanju javnog zdravlja, što bi u konačnici moglo značajno pridonijeti istraživanju prevencije, promocije i zaštite zdravlja stanovništva u XXI. stoljeću

Reference intervals for six salivary cortisol measures based on the Croatian Late Adolescence Stress Study (CLASS)

IN ENGLISH: INTRODUCTION: The aim of this nested study is to provide the reference intervals for already published measurements of salivary cortisol from the Croatian Adolescence Stress Study (CLASS). MATERIAL AND METHODS: A total of 969 individuals (372 males and 597 females) were included in the reference sample (age range: 18-21 years). Salivary cortisol concentrations were determined by the enzyme immunoassay (LUCIO-Medical ELISA Salivary Cortisol Kit, Nal von Minden, Germany) in the Department of Medical Laboratory Diagnostics, University Hospital Split. Nonparametric statistics were used for calculating the reference intervals (RIs) and 90% confidence intervals (90% CIs). RESULTS: The lower limits of RIs determined by the direct method were higher in females (> 10%) than in males for the cortisol concentrations at awakening (SCC0), 30 to 45 after awakening (SCC30-45) and at bedtime (SCCbedtime). The upper limits of RIs for the SCCbedtime were higher (> 10%) in males than in females. Females also had higher upper limits of RIs for the cortisol awakening response (CAR) and the diurnal cortisol slope (DCS) and higher lower limits of RIs for the CAR and the area under the curve with respect to ground (AUCG). The lower limits of RIs for the DCS were higher in males than in females. CONCLUSIONS: Obtained reference values open the arena for introducing salivary bioscience in Croatian clinical laboratory practice and provide important data for better understanding of gender differences in adaptation to stress during late adolescence. --------------- IN CROATIAN: UVOD: Cilj ove studije bio je definirati referentne intervale za objavljene mjere salivarnog kortizola iz Hrvatskog istraživanja stresa u kasnoj adolescenciji (CLASS). MATERIJALI I METODE: Ukupno 969 ispitanika (372 djevojaka i 597 mladića) uključeno je u referentni uzorak (18-21 godine starosti). Koncentracije salivarnog kortizola izmjerene su imunokemijskom ELISA metodom (LUCIO-Medical ELISA Salivary Cortisol Kit, Nal von Minden, Germany) u Zavodu za medicinsko laboratorijsku dijagnostiku Kliničkog bolničkog centra Split. Referentni intervali (RI) i 90%-tni intervali pouzdanosti (90% CI) izračunati su neparametrijskim metodama. REZULTATI: Donja granica referentnog intervala određena izravnom metodom veća je kod djevojaka (>10%) nego mladića za koncentracije kortizola u sve tri vremenske točke (po buđenju, 30-45 minuta nakon buđenja i neposredno prije lijeganja). Gornja granica referentnog intervala za koncentracije salivarnog kortizola neposredno prije lijeganja veća je kod mladića (>10%) u odnosu na djevojke. Također, djevojke imaju veću gornju granicu referentnog intervala za odgovor kortizola na buđenje (engl. cortisol awakening response, CAR) i cirkadijano smanjenje kortizola (engl. diurnal cortisol slope, DCS) te veću donju granicu referentnog intervala za CAR i površinu ispod krivulje u odnosu na nultu razinu (engl. area under the curve with respect to ground, AUCG). Indeks cirkadijanog smanjenja kortizola, DCS je u donjoj granici referentnog intervala veći kod mladića u odnosu na djevojke. ZAKLJUČCI: Objavljeni referentni intervali mjera salivarnog kortizola važni su zbog uvođenje salivarnih analiza u rutinsku kliničku praksu u RH i pridonose detaljnijim spoznajama o spolnim razlikama u adaptacijskim mehanizmima na stresne poticaje tijekom kasne adolescencije

Skeletal Remains from Late Roman Period: »As Old as Diocletian\u27s Palace«

In 2000, human skeletal remains were discovered in Split (Croatia). As archaeologists confirmed, it was an ancient skeleton accompanied by ceramics and bracelet characteristic for late Roman period whose possible violent death was excluded. The bone sample was radiocarbon dated by AMS to 1750 years. DNA was successfully extracted from the bone sample and subsequently typed using mt DNA and STR systems. The metal content was determined by atomic absorption spectrometry (AAS) in flame mode. Mercury concentration was determined by direct consecutive measures taken with a mercury analyzer. According to our results, we consider that the bones could belong to the one of the last citizens of the Diocletian’s Palace

Nasljedni karcinom dojke i jajnika - iskustva Kliničkog bolničkog centra Split

Aim: To investigate the clinical and pathohistological tumor characteristics, treatment, and treatment outcomes in patients with hereditary breast and ovarian cancer who were diagnosed, treated, and monitored at the University Hospital of Split from October 1999 to April 2021. Methods: The data were collected retrospectively from the medical history of 15 patients. They included the patient’s age at diagnosis, family history of malignancies, histological subtype, grade, breast cancer immunophenotype, stage of disease, status and types of BRCA mutations, type of surgical and oncological treatment, the specifics of metachronous bilateral breast cancers, the specifics of synchronous breast and ovarian cancers, and the outcome of treatment through overall survival (OS). Results: The median age of patients at the time of diagnosis of breast cancer was 53 years, and for ovarian cancer it was 56 years. A positive family history was confirmed in 13 patients (87%). All ovarian cancer patients had a high-grade serous histologic type, most often diagnosed in FIGO stages III and IV. Breast cancers were most commonly diagnosed in stages IA and IIA, with equally represented triple-negative and luminal immunophenotypes. The most common mutation was BRCA1 c.5266dup. The median OS of our patients was not reached. Conclusion: The clinical features of patients, pathohistological characteristics of tumors, and treatment outcomes in our study population are comparable with reports in the literature, respecting the specifics of different nations and races.Cilj: Istražiti kliničke osobitosti, patohistološke karakteristike tumora, način i ishode liječenja bolesnica s nasljednim karcinomom dojke i jajnika koje su dijagnosticirane, liječene i praćene u Kliničkom bolničkom centru Split od listopada 1999. do travnja 2021. godine. Metode: Podatci su prikupljeni retrospektivno iz povijesti bolesti 15 bolesnica. Uključivali su dob bolesnica kod dijagnoze bolesti, obiteljsku anamnezu za zloćudne bolesti, histološki podtip, gradus, imunofenotip karcinoma dojke, stadij bolesti, status i tip BRCA mutacija, osobitosti kirurškog i onkološkog liječenja, specifičnosti metakrono nastalih bilateralnih karcinoma dojke, specifičnosti sinkrono nastalih karcinoma dojke i jajnika te ishod liječenja kroz ukupno preživljenje. Rezultati: Medijan dobi bolesnica u trenutku dijagnoze raka dojke bio je 53 godine, a za karcinom jajnika 56 godina. Pozitivna obiteljska anamneza potvrđena je u 13 (87%) bolesnica. Karcinom jajnika je kod svih bolesnica bio seroznog papilarnog histološkog podtipa visokog gradusa i najčešće dijagnosticiran u FIGO stadiju III i IV. Karcinom dojke je najčešće dijagnosticiran u stadiju IA i IIA, jednake zastupljenosti trostruko negativnog i luminalnog imunofenotipa. Najčešća mutacija je bila BRCA1 c.5266dup. Medijan ukupnog preživljenja naših bolesnica nije dosegnut. Zaključak: Kliničke osobitosti bolesnica, patohistološke karakteristike tumora kao i ishodi liječenja u našoj studijskoj populaciji su usporedivi s izvješćima iz literature, respektirajući specifičnosti različitih naroda i rasa

Skeletal Remains from Late Roman Period: »As Old as Diocletian\u27s Palace«

In 2000, human skeletal remains were discovered in Split (Croatia). As archaeologists confirmed, it was an ancient skeleton accompanied by ceramics and bracelet characteristic for late Roman period whose possible violent death was excluded. The bone sample was radiocarbon dated by AMS to 1750 years. DNA was successfully extracted from the bone sample and subsequently typed using mt DNA and STR systems. The metal content was determined by atomic absorption spectrometry (AAS) in flame mode. Mercury concentration was determined by direct consecutive measures taken with a mercury analyzer. According to our results, we consider that the bones could belong to the one of the last citizens of the Diocletian’s Palace

Analysis of leptin, adiponectin and adiponectin gene polymorphism and leptin receptor in obese children and adolescents

Background: The aim of this study was to determine serum levels of leptin and adiponectin of obese children to identify the influence of leptin receptor gene polymorphisms on leptin resistance and leptin levels, as well as the association between the polymorphisms of adiponectin gene and adiponectin levels.Materials and methods: A case-control study comparing a study group of 74 obese children (age 13.34±2.60 years) to a normal weight-age matched (age 13.39±2.64 years) control group of 69 children. In both groups, body mass index (BMI) and waist/hip circumference, systolic and diastolic blood pressure were measured. Also, the leptin and adiponectin levels, as well as glucose and lipid metabolism parameters, and highly sensitive C-reactive protein (hs-CRP) were measured. Insulin sensitivity was evaluated using fasting insulinemia and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR). All subjects were tested for gene-tic polymorphisms in LEPRQ223R (rs1137101), ADIPOQ G276T (rs1501299) and ADIPOT45G (rs2241766).Results: The phenotypes of the obese children study group were significantly higher than in the control group in weight, BMI, waist/hip circumferences and systolic blood pressure (SBP) (P<0.001). We confirmed that in obese children the levels of leptin in the blood are increased and levels of adiponectin are decreased (P<0.001). The differences of the genotype distributions of leptin receptor (LEPRQ223R) and adiponectin (ADIPOG276T and ADIPOT45G) gene polymorphisms in the study group of obese chil-dren and a control group was not observed.Conclusion: In this study, we demonstrated increased leptin level and significantly decreased level of adiponectin in the obese children group compared with the control group. The results of the analysis of glucose metabolism and lipidogram between the two groups showed that insulin, HOMA-IR, and triglycerides, as well as hsCRP were increased and significantly different in the group of obese children compared to the control group, as expected. However, by including a significantly larger number of tested and control samples of both sexes and age-specific groups, with a larger number of tested SNPs, the genes investigated in this study would probably give better insight into a multicomplex disease such as obesity

Shiga toxin receptor Gb3Cer/CD77:tumor-association and promising therapeutic target in pancreas and colon cancer

BACKGROUND: Despite progress in adjuvant chemotherapy in the recent decades, pancreatic and colon cancers remain common causes of death worldwide. Bacterial toxins, which specifically bind to cell surface-exposed glycosphingolipids, are a potential novel therapy. We determined the expression of globotriaosylceramide (Gb3Cer/CD77), the Shiga toxin receptor, in human pancreatic and colon adenocarcinomas. METHODOLOGY/PRINCIPAL FINDINGS: Tissue lipid extracts of matched pairs of cancerous and adjacent normal tissue from 21 pancreatic and 16 colon cancer patients were investigated with thin-layer chromatography overlay assay combined with a novel mass spectrometry approach. Gb3Cer/CD77 was localized by immunofluorescence microscopy of cryosections from malignant and corresponding healthy tissue samples. 62% of pancreatic and 81% of colon adenocarcinomas showed increased Gb3Cer/CD77 expression, whereas 38% and 19% of malignant pancreas and colon tissue, respectively, did not, indicating an association of this marker with neoplastic transformation. Also, Gb3Cer/CD77 was associated with poor differentiation (G>2) in pancreatic cancer (P = 0.039). Mass spectrometric analysis evidenced enhanced expression of Gb3Cer/CD77 with long (C24) and short chain fatty acids (C16) in malignant tissues and pointed to the presence of hydroxylated fatty acid lipoforms, which are proposed to be important for receptor targeting. They could be detected in 86% of pancreatic and about 19% of colon adenocarcinomas. Immunohistology of tissue cryosections indicated tumor-association of these receptors. CONCLUSIONS/SIGNIFICANCE: Enhanced expression of Gb3Cer/CD77 in most pancreatic and colon adenocarcinomas prompts consideration of Shiga toxin, its B-subunit or B-subunit-derivatives as novel therapeutic strategies for the treatment of these challenging malignancies

Zajednički poslijediplomski doktorski studij zdravstvenih znanosti

Ovaj doktorski studij omogućio bi studentima potrebnu razinu znanja, vještina i stavova temeljenih na interdisciplinarnom pristupu u poimanju javnog zdravlja, što bi u konačnici moglo značajno pridonijeti istraživanju prevencije, promocije i zaštite zdravlja stanovništva u XXI. stoljeću