The TP53 gene polymorphisms and survival of sporadic breast cancer patients.

The TP53 gene polymorphisms, Arg72Pro and PIN3 (+16 bp), can have prognostic and predictive value in different cancers including breast cancer. The aim of the present study is to investigate a potential association between different genotypes of these polymorphisms and clinicopathological variables with survival of breast cancer patients in Croatian population. Ninety-four women with sporadic breast cancer were retrospectively analyzed. Median follow-up period was 67.9 months. The effects of basic clinical and histopathological characteristics of tumor on survival were tested by Cox's proportional hazards regression analysis. The TNM stage was associated with overall survival by Kaplan-Meier analysis, univariate, and multivariate Cox's proportional hazards regression analysis, while grade was associated with survival by Kaplan-Meier analysis and univariate Cox's proportional hazards regression analysis. Different genotypes of the Arg72Pro and PIN3 (+16 bp) polymorphisms had no significant impact on survival in breast cancer patients. However, in subgroup of patients treated with chemotherapy without anthracycline, the A2A2 genotype of the PIN3 (+16 bp) polymorphism was associated with poorer overall survival than other genotypes by Kaplan-Meier analysis (P = 0.048). The TP53 polymorphisms, Arg72Pro and PIN3 (+16 bp), had no impact on survival in unselected sporadic breast cancer patients in Croatian population. However, the results support the role of the A2A2 genotype of the PIN3 (+16 bp) polymorphism as a marker for identification of patients that may benefit from anthracycline-containing chemotherapy.

TP53 gene polymorphisms and breast cancer in Croatian women: a pilot study.

A case-control retrospective association study was conducted to investigate a possible association of the TP53 polymorphisms, Arg72Pro and PIN3 (+16bp), with sporadic breast cancer in Croatian women. Ninety-five women with breast cancer and 108 age-matched healthy women were analyzed. Arg72Pro polymorphism was detected by TaqMan essay. For designation of PIN3 (+16bp) polymorphism DNA amplification was performed by the polymerase chain reaction (PCR) while the PCR products were detected by capillary electrophoresis. Homozygous genotype of minor allele of the PIN3 (+16bp) polymorphism was associated with sporadic breast cancer (OR = 2.15, 95% confidence interval [CI] 1.80-2.56, p = 0.006). For Arg72 polymorphism, the odds ratio for breast cancer of ArgPro versus reference genotype ArgArg was 0.55 (95% CI 0.30-1.02, p = 0.039) suggesting the protective effect. Although different haplotypes did not influence the susceptibility to the disease, the joint occurrence of genotype combination ProPro/A2A2 frequent in cases, was associated with sporadic breast cancer (OR = 2.20, 95% CI 1.89-2.56, p = 0.021). The study provides evidence of the association of the TP53 gene polymorphisms Arg72Pro and PIN3 (+16bp) with sporadic breast cancer in the Croatian population.

Inbreeding and learning disability in Croatian island isolates.

The aim of this study was to investigate the prevalence of learning disability (LD) in isolate populations with different inbreeding coefficients (F). Prevalence of LD and F were determined in 10 villages from five Croatian islands: Brac, Hvar, Korcula, Lastovo and Susak. For the purpose of this study, LD was defined as the inability to attend the public school system. As the elementary schools (grade 1-8) in the place of the study are both public and compulsory, the assessment of child's inability to attend the school is performed at the age of six. This is required by all children in the country based on standard set of tests of cognitive performance defined by the Ministry of Education and Culture of the Republic of Croatia. The average inbreeding coefficients in each village population (F) were estimated in a random sample of 20-30% adults in each of the 10 villages based on 4 ancestral generations and using Wright's path method. Prevalence of LD ranged from 0.43% to 2.47%, and the inbreeding coefficients ranged from 0.8% to 4.9%. The Pearson's correlation coefficient between F and LD prevalence was 0.80 (p < 0.01). Although the relative risk per 5% inbreeding appeared very high (about 10), the absolute risk only increased from 0.18% to 1.77%. The genetic effect of inbreeding (GEI) was approximately 0.69% and the population-attributable fraction 76.6%. A review of the literature and the results of this study lead to a conclusion that a very large number of predominantly recessive genetic factors might mediate the genetic susceptibility to various forms of LD in these populations.

The evidence of mtDNA haplogroup F in a European population and its ethnohistoric implications.

Mitochondrial DNA polymorphism was analysed in a sample of 108 Croatians from the Adriatic Island isolate of Hvar. Besides typically European varieties of human maternal lineages, haplogroup F was found in a considerable frequency (8.3%). This haplogroup is most frequent in southeast Asia but has not been reported before in Europe. The genealogical analysis of haplogroup F cases from Hvar suggested founder effect. Subsequent field work was undertaken to sample and analyse 336 persons from three neighbouring islands (Brac, Korcula and Krk) and 379 more persons from all Croatian mainland counties and to determine if haplogroup F is present in the general population. Only one more case was found in one of the mainland cities, with no known ancestors from Hvar Island. The first published phylogenetic analysis of haplogroup F worldwide is presented, applying the median network method, suggesting several scenarios how this maternal lineage may have been added to the Croatian mtDNA pool.

A signal, from human mtDNA, of postglacial recolonization in Europe.

Mitochondrial HVS-I sequences from 10,365 subjects belonging to 56 populations/geographical regions of western Eurasia and northern Africa were first surveyed for the presence of the T-->C transition at nucleotide position 16298, a mutation which has previously been shown to characterize haplogroup V mtDNAs. All mtDNAs with this mutation were then screened for a number of diagnostic RFLP sites, revealing two major subsets of mtDNAs. One is haplogroup V proper, and the other has been termed "pre*V," since it predates V phylogenetically. The rather uncommon pre*V tends to be scattered throughout Europe (and northwestern Africa), whereas V attains two peaks of frequency: one situated in southwestern Europe and one in the Saami of northern Scandinavia. Geographical distributions and ages support the scenario that pre*V originated in Europe before the Last Glacial Maximum (LGM), whereas the more recently derived haplogroup V arose in a southwestern European refugium soon after the LGM. The arrival of V in eastern/central Europe, however, occurred much later, possibly with (post-)Neolithic contacts. The distribution of haplogroup V mtDNAs in modern European populations would thus, at least in part, reflect the pattern of postglacial human recolonization from that refugium, affecting even the Saami. Overall, the present study shows that the dissection of mtDNA variation into small and well-defined evolutionary units is an essential step in the identification of spatial frequency patterns. Mass screening of a few markers identified using complete mtDNA sequences promises to be an efficient strategy for inferring features of human prehistory.

Short tandem repeat (STR) variation in eight village populations of the island of Korcula (Croatia).

The aim of this study was to analyse short tandem repeat (STR) variation using the data on nine loci (D3S1358, vWA, FGA, THO1, TPOX, CSF1PO, D5S818, D13S317, D7S820) in the populations from eight villages on the island of Korcula, Croatia, in order to analyse its genetic and population structure. The analysis of STR data in this study indicated an appreciable degree of genetic homogeneity among the studied village populations on the island, even though a so-called 'east-west dichotomy' and differentiation between the inhabitants of the most recent settlement and the remaining ones was indicated with respect to the loci CSF1PO and TPOX, respectively. The validity of STR markers in assessing genetic structure of small populations and especially in determining the relationships among geographically closely related but reproductively isolated groups remains to be further evaluated, especially in terms of a larger number of studied loci in order to possibly find specific markers useful for resolving genetic patterns of variability at regional levels.

A large cross-sectional study of health attitudes, knowledge, behaviour and risks in the post-war Croatian population (the First Croatian Health Project).

As the liberation of occupied Croatian territories ended the war in the country in 1995, the Ministry of Health and Croatian Health Insurance Institute have agreed to create the new framework for developing a long-term strategy of public health planning, prevention and intervention. They provided financial resources to develop the First Croatian Health Project, the rest of the support coming from the World Bank loan and the National Institute of Public Health. A large cross-sectional study was designed aiming to assess health attitudes, knowledge, behaviour and risks in the post-war Croatian population. The large field study was carried out by the Institute for Anthropological Research with technical support from the National Institute of Public Health. The field study was completed between 1995-1997. It included about 10,000 adult volunteers from all 21 Croatian counties. The geographic distribution of the sample covered both coastal and continental areas of Croatia and included rural and urban environments. The specific measurements included antropometry (body mass index and blood pressure). From each examinee a blood sample was collected from which the levels of total plasma cholesterol (TC), triglycerides (TG), HDL-cholesterol (High Density Lipoprotein), LDL-cholesterol (Low Density Lipoprotein), lipoprotein Lp(a), and haemostatic risk factor fibrinogen (F) were determined. The detailed data were collected on the general knowledge and attitudes on health issues, followed by specific investigation of smoking history, alcohol consumption, nutrition habits, physical activity, family history of chronic non-communicable diseases and occupational exposures. From the initial database a targeted sample of 5,840 persons of both sexes, aged 18-65, was created corresponding by age, sex and geographic distribution to the general Croatian population. This paper summarises and discusses the main findings of the project within this representative sample of Croatian population.

Anthropogenetical analysis of abnormal human alpha-globin gene cluster arrangement on chromosome 16.

An earlier study of human globin gene polymorphism in two Adriatic islands of Olib and Silba showed an abnormal arrangement of alpha-globin genes in two different individuals. The next step was to determine the degree of the kinship relationship between the two probands, one with a deleted and another with triplicated alpha-globin gene on the island Silba, and to determine the stability of this disorder through generations. We reviewed the parish registers (Status Animarum) of the island of Silba, dating from the year 1527, and constructed family trees for the two probands. Restriction endonuclease mapping was performed to study the arrangement of the alpha-globin genes in the offspring of our probands. A total of 183 ancestors completed the two family trees. The kinship relationship between them was established in the 5th, 6th, and 7th generation. The analysis of alpha-globin genes in the offspring of our probands showed the triplicated alpha-globin genes in two persons. We also found alpha-globin gene triplication in other three relatives. We did not find any deleted alpha-globin genes. We determined the kinship relationship between the two probands, one with deleted and the other with triplicated alpha-globin genes. This finding enabled us to determine the stability of this gene disarrangement through generations. It also showed new possibilities in anthropogenetic research, by combining the analyses of parish registers with those of modern genetic methods, such as restriction endonuclease mapping.

MtDNA haplogroups in the populations of Croatian Adriatic Islands.

The number of previous anthropological studies pointed to very complex ethnohistorical processes that shaped the current genetic structure of Croatian island isolates. The scope of this study was limited to the general insight into their founding populations and the overall level of genetic diversity based on the study mtDNA variation. A total of 444 randomly chosen adult individuals from 32 rural communities of the islands of Krk, Brac, Hvar and Korcula were sampled. MtDNA HVS-I region together with RFLP sites diagnostic for main Eurasian and African mtDNA haplogroups were analysed in order to determine the haplogroup structure. The most frequent haplogroups were "H" (27.8-60.2%), "U" (10.2-24.1%), "J" (6.1-9.0%) and "T" (5.1-13.9%), which is similar to the other European and Near Eastern populations. The genetic drift could have been important aspect in history, as there were examples of excess frequencies of certain haplogroups (11.3% of "I" and 7.5% of "W" in Krk, 10.5% of "HV" in Brac, 13.9% of "J" in Hvar and 60.2% of "H" in Korcula). As the settlements on the islands were formed trough several immigratory episodes of genetically distant populations, this analysis (performed at the level of entire islands) showed greater genetic diversity (0.940-0.972) than expected at the level of particular settlements.

STR polymorphisms in the population of the island of Hvar.

The aim of this study is to analyze short tandem repeat (STR) variation using data on 9 loci (D3S1358, VWA, FGA, THO1, TPOX, CSF1PO, D5S818, D13S317, D7S820) from the subpopulations of 6 villages on the island of Hvar, Croatia. The STR data help us to analyze the genetic structure of Hvar. The analysis of STR data in this study indicated genetic homogeneity among the village subpopulations on Hvar and the lack of the so-called east-west dichotomy, which had been indicated by some previous multidisciplinary anthropological studies. The observed value of GST (0.030) is most probably a consequence of high STR mutation rates, which produce a high level of within-group (village) diversity relative to total diversity of the population. The validity of STR markers in assessing genetic structure of small populations and especially in determining the relationships among closely related and reproductively isolated groups remains to be further evaluated.

VNTR DNA variation in the population of the island of Hvar, Croatia.

The aim of this study was to investigate hypervariable DNA polymorphisms in the Croatian population. Two VNTR loci, D7S22 and D12S11, were studied in a sample of 68 inhabitants of the western and eastern regions of the island of Hvar. Binned allele frequencies and heterozygosity were calculated for the two regions and measures of genetic kinship and genetic distance were computed between the two regions and between each of these regions and nine world populations. The relatively large genetic distance between the two regions of the island of Hvar (0.0353) may reflect historical immigrations from the mainland and interesting peculiarities of the populations that arrived from the Balkan peninsula.

HLA class II gene and haplotype diversity in the population of the island of Hvar, Croatia.

The DRB1, DRB3, DRB5, DQA1 and DQB1 allele polymorphisms were analysed in 3 western and 3 eastern villages of the island of Hvar using PCR-SSOP method and 12th International Workshop primers and probes. Three DQB1 alleles (*0304, *0305, *0607) detected in the population of the island of Hvar (HP) have not yet been observed in general Croatian population (GCP). Significant differences were observed between two regions of Hvar for: a) DRB1*0701 allele (p < 0.001), b) DQA1*0201 allele (p < 0.01), and c) DRB1*0101-DQA1*0101-DQB1*0501 haplotypic association (p < 0.05). Two unusual haplotypic associations, which have not yet been described in general Croatian population (GCP), DRB1*0101-DQA1*0102-DQB1*0501 and DRB1*1501-DQA1 *0102-DQB1*0604 were observed in the population from the island of Hvar (HP). Measures of genetic kinship and genetic distances revealed isolation and clusterization which coincides with the known ethnohistorical, as well as biological and biocultural data obtained from a series of previous investigations. The five studied village subpopulations formed two clusters (East-West) to which the far eastern village (with the highest rii of 0.0407) joined later, thus indicating possible impact of historical immigrations from the mainland.

Application of HLA class II polymorphism analysis to the study of the population structure of the Island of Krk, Croatia.

The population structure of the northern Adriatic island of Krk, Croatia, was studied using PCR methodology and nonradioactive oligonucleotide hybridization for the analysis of HLA-DRB1, DRB3, DRB4, DRB5, DQA1, and DQB1 polymorphisms. Allele frequencies, genetic kinship (R), and genetic distances (E2) were computed, and correlations between distance (genetic, linguistic, geographic) and kinship (migration) matrices were examined. The results, reflecting past (micro-) evolutionary processes, indicate that ethnohistorical and sociocultural events rather than geographic distances are the primary determinants of today's population structure of the island.

The aging process--an analysis of the latent structure of body morphology (in males).

The morphological characteristics (20 anthropometric variables) of a total of 2,351 examinees (from the age of 18 to 90) were analyzed by a model of the principal components of the factor analysis. Four factors were extracted that explain 71.4% of the total variance. The factors-"general body voluminosity", "subcutaneous fat tissue", "longitudinal body dimensionality" and "upper body voluminosity"-were analyzed within the context of their appearance in different age-determined cohorts. The differences between cohorts (groups per ten years of age) were studied by the canonical discriminant analysis. The first two discriminant functions (describing mostly the variability of cohorts-96.11%) indicate a constant decrease of body and sitting height, and an increase of upper body voluminosity till the fourth age cohort, which is the most crucial one in the change of latent morphological structure. Results of the correct classification of cohort members show that only 48.45% of probands were correctly placed (the best classification determined was in the age between 46 and 55 years) indicating that in males, at least three different groups exist according to the specificity of morphological aging in human organisms.

Estimation of radioactive calcium absorption based on expanding Ca-exchangeable space.

Anthropological interest in calcium metabolism is partially connected with the genetic basis of skeletal metabolism that could provide additional optimism for people suffering from osteoporosis. The present investigation is aimed at deal with a of model describing Ca intestinal absorption, suitable for other descriptions currently investigated in anthropology (migration and gene flow). The purpose of this investigation was to establish features of the reflection of the Ca isotopic absorption tests in serum using the concept of an expanding Ca-space and to assess the reliability of the estimation of absorption (ABS) from one serum sample. Double tracer method, using 85Sr as the iv tracer, was used as the referent. The study included 100 subjects (49 females, 51 males) of which 34 were considered healthy. The mean value of ABS was 68 (+/-17.45 SD) % of dose. Serum radioactivity during the 24 hours following oral administration of 47Ca (together with 100 mg elemental Ca), o.s(t), from five blood samples was determined. The relationship between o.s(t) and ABS was found to be connected to an expanding Ca-space, V(t): ABS = o.s(t)V(t) = 26 + 11.8t0.35 o.s(t). After the absorption process was finished, an approximate value of V(t) is (100/S1)(t-x)b. The constants "S1" and "b" describe power function of iv s(t), whereas parameter "x" represents delay of V(t) Ca-space at oral administration in comparison to the iv administration. Statistical models selected on the basis of their low values of the SEE (from 8.71 to 8.90% of dose) include body surface index or body weight index (BW0.425). The CV expressed as (SEE/mean x ABS(Ca/Sr) x 100 were about 13% and the observed greatest difference between the estimated and measured ABS(Ca/Sr) was 20% of dose. The authors believe that models presented permit, to a limited extent, comparison of results obtained by different procedures. The results, according to variability of the sample of subjects, permit judgment on upper limits of error at estimation of absorption from one blood sample radioactivity.

Anthropogenetic variability of populations of the north-eastern part of the Slavonia and Baranya Region--Croatia.

In this work we studied the anthropogenetic structure of eight populations in the north-eastern part of the Slavonia-Baranya region. The results are based on the analysis of ABO and Rh systems of erythrocytic antigens i.e. analysis of A1, A2, B and O genes, and in Rh systems D and d genes. The analysis was carried out on 7,860 grown-ups chosen by a random sample method among blood donors in Clinical Hospital in Osijek. The gene frequencies, "genetic distances", coefficients of kinship and heterozygosity have been calculated for examined populations. The results show that the present anthropologenetic population structure reflects the common origin of the examined populations, but also that the numerous changes throughout their ethno-history, caused first of all by population migrations, which is one of the basic mechanisms that have shaped the population structure of the north-eastern part of Slavonia-Baranya region.

Serogenetic analysis in the study of the population structure of the eastern Adriatic (Croatia).

The anthropogenetic structure of six island and peninsular populations (Brac, Hvar, Korcula, Peljesac, Silba, and Olib) of the eastern Adriatic, Croatia, is analyzed on the basis of the study of four different erythrocyte antigen systems or groups (ABO, Rhesus, Kell-Celano, P) and two erythrocyte isoenzyme systems (ACP, ESD). The average sample size was 555 individuals. Allele frequencies, genetic distances, and gene diversity values were computed. The results indicate that all the populations in question have preserved their separate characteristics over the course of their (micro)evolution to the present day; this is especially noticeable for the island populations of Korcula and Olib, as these are distinguished from the other four populations by a greater degree of isolation. Today's genetic structure of the six populations can be explained through the existing historical and cultural data for the region in question, which indicate that over the course of their ethnohistory they were all influenced by significant waves of immigration and selective emigrations that must have greatly shaped their present-day population structure.

Population structure of the Island of Korcula, Croatia.

Serogenetic data on 16 systems and migration data from birthplaces of parent and offspring, collected from 949 inhabitants of 8 vilages on the island of Korcula in the eastern Adriatic (Croatia), were analysed. Geographic distance is not a strong determinant of the population structure on the island. The pattern of the genetic variation now observed derives mainly from history and migration behavior. © 1993 Wiley-Liss, Inc.

Population structure of the Peljesac Peninsula, Yugoslavia.

We gathered serogenetic and parent-offspring migration data from 604 residents of 7 villages on the Peljesac peninsula in southern Yugoslavia. A variety of population genetics and multivariate statistics models and procedures give a concordant picture of the population structure of this region. Extensive migration is the dominant microevolutionary force patterning the variation seen today. Multiple population bottlenecks have also occurred over the past few centuries as a result of disease, famine, war, economic failure, and founder events, making it likely that genetic drift has been an important factor in the history of this population system.