ESHG Posters 9

VNTR Cassette Sequence Diversity In The European R408W-1.8 And R408W-2.3 Phenylketonuria Mutation Lineages

D. T. Croke¹, O. Tighe¹, C. O'Neill², P. Mayne²;
¹Royal College of Surgeons in Ireland, Dublin, IRELAND, ²The Children's Hospital, Temple Street, Dublin, IRELAND.

R408W, the predominant European Phenylalanine Hydroxylase (PAH) mutation in Phenylketonuria (PKU), has arisen by recurrent mutation on chromosomes of haplotypes 2.3 and 1.8. The R408W-2.3 and R408W-1.8 mutations exhibit west-east and east-west clinal distributions across Europe respectively. Crucial evidence for the origin of R408W by recurrent mutation was provided by a previous report [Byck et al. Hum. Molec. Genet. 1994; 3: 1675-1677.], which demonstrated that R408W had arisen on a rare VNTR cassette sequence variant of haplotype 1.8. We have investigated VNTR sequence variation across a range of European populations. DNA samples (n=141) from thirteen European regions were obtained (with appropriate ethical approval) in collaboration with the members of the European PAH VNTR cassette sequence variation study group. VNTR alleles were amplified by PCR, gel-purified and cycle-sequenced in forward and reverse orientation. Cassette sequences were identified according to the standard nomenclature. VNTR-3 alleles exhibited a single cassette organisation (a2-b2-c1) common to all chromosomes whether of wild-type or R408W mutant genotype. In contrast, wild-type VNTR-8 alleles had two different cassette structures represented within the study cohort at roughly equal frequency, namely (a1)5-b3-b2-c1 and (a1)5-b5-b2-c1. R408W-1.8 chromosomes exhibited the (a1)5-b5-b2-c1 VNTR structure alone but were associated with different STR alleles compared to wild-type 1.8 chromosomes. These data confirm the earlier suggestion that recurrent mutation gave rise to R408W mutations on different haplotype backgrounds in Europe and raise interesting questions regarding the relative ages of the R408W-1.8 and -2.3 lineages.

H. Rangel-Villalobos¹, L. Sandoval², B. Ibarra², J. Ana Rebeca², A. Gonzalez-Martin³, L. Figuera²;
¹Centro Universitario de la Cienega (CUCI-UdeG), Ocotlán, Jalisco, MEXICO, ²Centro de Investigación Biomédica de Occidente (IMSS), Guadalajara, Jalisco, MEXICO, ³Universidad Autonoma del Estado de Hidalgo, Hidalgo, MEXICO.

The non-pseudoautosomal region of the Y-chromosome constitutes a genetic record easily interpretable to obtain valuable anthropological information about the history of worldwide populations. Two bi-allelic loci (YAP and DYS199) and five STRs (DYS19, 389a, 390, 391 and 393) of the non-pseudoautosomal region of the Y-chromosome were analyzed in males from the largest and most widely distributed population in Mexico (Mestizos) and from three Mexican Amerindian tribes: Huichols, Purepechas and Tarahumaras. The allelic distribution of all seven loci was established and it was pairwise compared between populations. For YAP locus, any significant difference (p> 0.05) was observed among all four populations. The Amerindian-specific allele DYS199-T was more frequent in Mexican tribes than in Mestizos, establishing the minimum Amerindian component in the Mestizo sample as 18.6%. Tarahumaras were peculiar by its diminished frequency for DYS199-T respecting to Purepechas and Huichols. Mexican Mestizos were different (p<0.05) to Huichols, Purepechas and Tarahumaras in five, four and two STRs, respectively. Eighty-eight different haplotypes were observed among the 156 haplotypes obtained. They were grouped in three haplogroups according to the markers YAP (+/-) and DYS199 (C/T):
-/C, -/T and +/C. The greater haplotype diversity (D) was observed in Mestizos (98.6 %) and the lower in Huichols (87.17 %). The haplotype variation of the Y-chromosome in Mexican populations was analyzed by AMOVA. The inter-population and intra-population variations were significant (p<0.0001) and constituted the 78.5% and 21.5%, respectively. We discuss our findings with previous results about the same populations using autosomal markers (Hum. Biol. 2000, 72: 983-995).

Human mitochondrial DNA control region sequence variations in Lithuanian population

D. Kasperaviciute, V. Kucinskas;
Human Genetics Centre of Vilnius University, Vilnius, LITHUANIA.

The Lithuanians and Latvians are the only two Baltic cultures that survived until today. There are conflicting anthropological findings regarding the process of neolithization in the Baltic region and the formation of the Baltic tribes. However, since neolithic period the native inhabitants of Lithuanian territory have not been replaced by any other ethnic group. Therefore the genetic characterization of the present day Lithuanians may shed more light on the early history of the Balts.
We have analyzed 120 DNA samples from two Lithuanian ethnolinguistic groups (Aukstaiciai and Zemaiciai) using direct sequencing of the first hypervariable segment (HVI) of the control region of the mitochondrial DNA and restriction enzyme digestion for polymorphic site 00073. On the basis of specific substitutions the obtained sequences were classified to mtDNA clusters defined by Richards et al. Sequences of almost all major European clusters (except X) were found in Lithuania. Haplogroup H was the most common mtDNA lineage reaching frequencies from ~40% in Zemaiciai to ~45% in Aukstaiciai. The second most prevalent haplogroup was U, comprising from 25% to 36% of the sequences in Aukstaiciai and Zemaiciai respectively. The frequencies of remaining haplogroups were from 2% to 10%. In general, the mtDNA lineages reflecting more ancient demographic expansion seem to be more frequent, which is compatible with anthropological findings that neolithization in the Baltic region has been largely indigenous process. However, the lineages reflecting the spread of early farmers from the Near East are also present, indicating different processes in the history of Lithuanian population.

Isonymy, inbreeding and isolation in San Miguel Island (Azores, Portugal): A surname study.

C. C. Branco, L. Mota-Vieira;
Genetic and Molecular Pathology Unit, Hospital of Divino Espirito Santo, Azores Islands, PORTUGAL.

Small islands constitute isolated populations that offer advantages in disease locus positioning and gene identification. Here we present the population structure of San Miguel island (131 530 inhabitants, 2001 Census), the biggest of nine islands of the Azorean Archipelago (Portugal). Our study was based on surname frequencies that were obtained from the most recent telephone list (2001). We identified 1 315 different surnames in a total of 27 621 subscribers. Eleven places, including the capital (Ponta Delgada) and other rural communities (Achada, Bretanha, Furnas, Ginetes, Maia, Nordeste, Rabo-de-Peixe, Povoação, Salga and Sete Cidades), were chosen according to population size and geographic isolation. Isonymy (I), inbreeding coefficient (Fst), coefficient of kinship between locations (Ri), Fisher’s a (a), Karlin-McGregor n (n) and Nei’s distance were calculated. Salga presents the highest values of isonymy (0.0576) and Fst (0.0144) and the lowest value of a (17.36). Sete Cidades presents the highest value of n (0.130). Moreover, 51% of Salga’s population and 52% of Sete Cidades’s population are represented by 6 and 8 surnames, respectively. These results demonstrate the effective isolation and high rates of emigration of these two places, located in opposite edges of the San Miguel island. In contrast, the capital shows the lowest values of isonymy (0.0128), Fst (0.0032) and n (0.0136) and the highest value of a (78.13). Our analysis suggests a high degree of inbreeding in rural communities of San Miguel island which often constitute a model for genetic mapping studies. (claudia.branco@clix.pt)

E. Samilchuk¹, I. Al-Suliman¹, E. Usanga², S. Al-Awadi¹;
¹Kuwait Medical Genetics Centre, Ministry of Health, KUWAIT, ²Faculty of Allied Health, Kuwait University, KUWAIT.

Glucose-phosphate dehydrogenase (G6PD) deficiency is caused by mutations in the G6PD gene. The clinical manifestation of G6PD deficiency may be influenced by a (TA)n polymorphisn in a promoter of the UDP-glucuronosyltransferase 1 (UDPGT1) gene. The (TA)7 allele is associated with increase in the incidence of neonatal hyperbilirubinaemia in G6PD deficient newborns. To analyze G6PD mutations and UDPGT1 polymorphism in G6PD deficient Kuwaitis, 1080 male blood donors were screened for G6PD activity. G6PD deficiency was identified in 70 (6.5%) individuals. Mutation analysis revealed the Mediterranean (C563T) mutation in 51 (73%), A- (G202A) in 10 (14%), Chatham (G1003A) in 5 (7.1%), Aures (T143C) in 1 (1.4%) of 70 G6PD deficient cases. In three (4.2%) cases mutations remain unknown. Both the Mediterranean and Aures mutations were associated with the C1131T polymorphism while the A- mutation was accompanied by the A376G substitution. UDPGT1 genotyping revealed the (TA)6/(TA)6 genotype in 27 (38.6%), (TA)6/(TA)7 in 31 (44.3%), (TA)7/(TA)7 in 11 (15.7%), and (TA)6/(TA)8 in 1 (1.4%) of 70 G6PD deficient cases. The frequencies of the (TA)6, (TA)7, and (TA)8 alleles among G6PD deficient Kuwaitis were 0.6143, 0.3786 and 0.0071, respectively. The rare (TA)8 allele was observed in a G6PD deficient individual with the A- mutation (both A- and (TA)8 are known to be of African origin).

Evidence of a founder effect for the tissue-nonspecific alkaline phosphatase (TNSALP) gene E174K mutation in hypophosphatasia patients

M. Herasse¹, M. Spentchian², A. Taillandier¹^,3, E. Mornet¹^,3;
¹Université de Versailles-Saint Quentin en Yvelines, Versailles, FRANCE, ²Hopital André Mignot, Versailles, FRANCE, ³Laboratoire SESEP, Versailles, FRANCE.

Hypophosphatasia is an inborn error of metabolism characterized by defective bone mineralization caused by a deficiency of liver-, bone- or kidney-type alkaline phosphatase due to mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene. The clinical expression of the disease is highly variable, ranging from stillbirth with poor mineralized skeleton to pathologic skeletal fractures which develop in late adulthood only. This clinical heterogeneity is due to the strong allelic heterogeneity in the TNSALP gene. Mutation E174K is the most frequent mutation in Caucasian patients and represents 8% of hypophosphatasia chromosomes. This mutation was found in patients from various geographic origins but was more frequent in the North of Western Europe. We therefore investigated the likelihood of its unique origine or the likelihood of a multiple origin due to recurrence of the mutation on several chromosomes. Three intragenic polymorphisms, S93S, 472+12delG and V505A were genotyped in patients carrying E174K and in normal unrelated individuals. The results show that all the E174K mutations are carried out by a common ancestral haplotype, also found at low frequency in normal and hypophosphatasia chromosomes. We conclude that the E174K mutation is the result of an ancestral mutation that ocurred on a single chromosome in the North of Western Europe and spread throughout the rest of Europe and into the New World as a result of migrations.

D. Ambrasiene, V. Kucinskas;
Human Genetics Centre, Vilnius University, Vilnius, LITHUANIA.

The human Y chromosome is uniparentally inherited and nonrecombining along most of its length. Short tandem repeat (STR) polymorphisms from the male specific part of Y chromosome have been already recognised to be highly valuable in human evolutionary studies and population genetics. Present-day Lithuanians represent the Baltic branch of the populations speaking languages of the Proto-Indo-European descent. Although Lithuanian population was formed under the pressure of various migration forces, its deep roots preserved the genetic composition of the forebears. Therefore, it is reasonable to analyse genetic differences among the Lithuanian ethnolinguistic groups as well as between Lithuanians and other European populations. We present results of the investigation of genetic structure of Lithuanian population by using Y-chromosomal microsatellite markers. We examined the allele and haplotype frequencies of the Y chromosome-specific STR systems (DYS19, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS385) in 57 males from two main Lithuanian ethnolinguistic groups: Aukstaiciai (A) and Zemaiciai (Z). PCR products were detected using capillary electrophoresis on the ABI PRISM 310. In total 44 different haplotypes were identified as a result of combining the 32 alleles of the 7 Y-linked systems containing 9 loci. 35 haplotypes were seen only once, five - twice, four - thrice. The most frequent allele of DYS19 was 15 (52.6%), DYS389-I - 13 (71.9%), DYS389-II - 30 (57.9%), DYS390 - 25 (49.1%), DYS391 - 11 (54.4%), DYS392 - 11 (59.5%), DYS393 - 13 (61.4%), DYS385 - 11/14 (43.9%). Our results were compared with the data of other European populations.

High resolution analysis of human Y-chromosome diversity in the Western Mediterranean area

F. Cruciani¹, P. Santolamazza¹, P. Moral², G. Vona³, R. La Fratta¹, V. Proietto¹, A. Torroni¹^,4, R. Scozzari¹;
¹Dipartimento di Genetica e Biologia Molecolare, Università "La Sapienza", Rome, ITALY, ²Departament de Biologia Animal, Universitat de Barcelona, Barcelona, SPAIN, ³Dipartimento di Biologia Sperimentale, Università di Cagliari, Cagliari, ITALY, ⁴Dipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ITALY.

Y chromosome variation was analyzed by surveying 33 biallelic markers in a sample of about 900 males belonging to 23 populations from the Western Mediterranean area. Some populations from the Middle East were also included for comparison. This survey revealed a total of 21 binary haplotypes which were combined with the data from 7 microsatellites to evaluate internal diversities and coalescence ages. The dissection of the YAP+ lineage in several distinct haplotypes showed that only a few derivative haplotypes are present in Europe. One of these haplotypes, defined by the mutation M81, is phylogenetically equivalent to the previously described haplogroup 25.2 (Scozzari et al. Hum Immunol 62:871, 2001). This haplotype is very common in North Western Africa and most likely originated in the Berbers of Morocco where it reaches frequencies of about 70%. The same haplotype is rare in Europe, the only exception being represented by the Spanish from the Pas Valley (43%), thus indicating a strong component of Northern African origin in that region. The haplotype defined by the M26 mutation resulted to be phylogenetically equivalent to the haplotype carrying the 11 repeats allele at the YCAIIb microsatellite. This haplotype is very frequent in Sardinia but is rare or absent in other regions. Overall this study demonstrates that the dissection of Y-chromosome variation into haplogroups/haplotypes with a more restricted geographic distribution can reveal important affinities between populations and provides new clues about their past interactions.

E. V. Shabrova¹, E. K. Khusnutdinova², A. I. Mikulich³, L. A. Tarskaia¹, S. A. Limborska¹;
¹Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, RUSSIAN FEDERATION, ²Institute of Biochemistry and Genetics, Ufa scientific center, Russian Academy of Sciences, Ufa, RUSSIAN FEDERATION, ³Institute of Arts, Ethnography and Folklore, Belarussian Academy of Science, Minsk, BELARUS.

Over the past years multilocus DNA fingerprinting have been widely applied for the genetic population studies on the different kinds of organisms. We have used this technique with M13 phage DNA as a hypervariable minisatellite probe to investigate 13 human populations from Eastern Europe and Asia. These populations belongs to three language families: Indo-European language family (Slavonic branch: Belarussian), Uralic language family (Finno-Ugric branch: Mari, Mordvinians, Udmurts, Komi), Altai language family (Turkic branch: Bashkirs, Tatars, Chuvashes, Yakuts).
The matrice of all the fragments with the sizes from 10,000-2,000 bp was constructed as a result of the analysis of restriction fragments patterns generated by hybridization. The set of individual patterns was presented as a binary matrice like "Object-Trait".
Level of the population differentiation was estimated according to Lynch (1).
Different statitical analyses (cluster analysis, multiple correspondence analysis, multidimensional scaling) were applied for the treatment of distance matrice obtained from the populations profiles of the fragments frequencies and binary populations matrices as well.
Two territorially separated Belarussian populations produced no regional differnces whereas four separated Bashkir populations appeared to be quite different.
On the whole the correlation between our results and the linguistic affinity data is not absolute.
1. Lynch M. (1990). The Similarity Index and DNA Fingerprinting. Mol. Biol. Evol. 7(5).

C. Deltas¹, E. Rossou¹, R. Mean¹, P. Koupepidou¹, C. Costi¹, M. Koptides¹, V. Atamian², A. Pierides³;
¹The Cyprus Institute of Neurology and Genetics, Nicosia, CYPRUS, ²Private Practice, Nicosia, CYPRUS, ³Dept of Nephrology, Nicosia, CYPRUS.

Familial Mediterranean fever (FMF) is an autosomal recessive disease of high prevalence in four ethnic groups, Non-Ashkenazi Jews, Armenians, Arabs and Turks. Typically it presents as acute episodes of periodic fever accompanied by abdominal pain, chest pain, or joint pain. The attack usually lasts from 12 to 72 hours, with arthralgia or arthritis often lasting longer. The most dangerous potential complication is amyloidosis that can lead to ESRF. About 40 mutations have been identified so far, some of them being very frequent. Founder effects have been postulated to be responsible for the high frequency of certain mutations in selected populations. Molecular investigation of the Cypriot population reveals that about 1:8 is a carrier of one of four mutations, E148Q being the most frequent (1:12). Among 87 MEFV chromosomes analysed, the results are: V726A 27.6%; F479L 21.8%; M694V 20.7%; E148Q 6.9%; M694I 2.3%; R761H 2.3%; Unknown 18.4%. Mutation F479L is rather rare in other populations. Preliminary evidence suggests that this frequent Cypriot mutation is associated with later age of onset of symptoms, the most debilitating of which is strong and frequent abdominal pain, with or without fevers and arthralgias. Despite the high frequuency of E148Q, only 6 of 53 patients carried it, supporting its mild nature. More than have of documented cases were diagnosed during the past year, after announcing the availability of molecular testing and providing evidence that FMF is not rare. Molecular testing is expected to assist further in identifying ambiguous cases, while general newborn screening is under consideration.

T. Reimand¹^,2, K. Õunap², R. Zordania³, T. Ilus², T. Talvik¹, M. Sitska²;
¹University of Tartu, Department of Pediatrics, Tartu, ESTONIA, ²Tartu University Clinics, Medical Genetics Center, Tartu, ESTONIA, ³Children Hospital of Tallinn, Tallinn, ESTONIA.

The aim of this study was to investigate the incidence of Down syndrome (DS) in Estonia.
Methods: The data about the children with DS were collected from genetic centers of Estonia, from database of Down Syndrome Supportive Groups, from institutions of disabled children and from the registers of family doctors/pediatricians. The study subjects were 196 DS patients born from 1990 to 2000. Three of them died before cytogenetic investigation was performed. Therefore 193 children with cytogenetically confirmed DS were included into the study.
Results: Regular trisomy was found in 173 cases (90%), translocation in 14 cases (7%) and mosaicism in 6 cases (3%). In one patient there was regular trisomy 21 and translocation between 13;14 chromosomes. Mosaicism and translocation at the same time (47,XY,t(7;21)(80%)/46,XY(20%)) was found in one child. Thirty percent of the mothers were older than 35 years.
In 1995 screening of chromosome anomalies for advanced maternal age (>35) was started. Therefore we divided the DS children into two groups: born in 1990-1994 and 1995-2000. The first group consist 108 patients (birth-rate: 89015). According to these data the incidence of DS was 1:824. In the second group DS was diagnosed in 85 children (birth-rate: 76767). According to the data of Medical Genetic Center in 32 cases of DS was diagnosed prenatally; in 31 cases the pregnancy was terminated. If those 31 DS children were born the incidence of DS in the second group would be 1:662.
In conclusion we may say that the provisional incidence of DS is 1:700.

Y-chromosome haplotype analysis in three Eastern Europe populations - Belorusia, Russia and Ukraine.

S. A. Kravchenko¹, S. A. Limborskaya², L. A. Livshits¹;
¹Institute of Molecular Biology and Genetics, Kiev, UKRAINE, ²Institute of the Molecular Genetics, Moskow, RUSSIAN FEDERATION.

Y-chromosomal microsatellite haplotypes are highly valuable in human evolutionary and human history populations studies. Distribution of Y-chromosome haplotypes for 5 microsatellite loci ( DYS393, DYS392, DYS391, DYS390 and DYS19 ) was investigated in three population- Belorussia, Russia and Ukraine to reconstruct the evolution of paternal lineage in populations with Slavic origin from East Europe.
To determine affinity between Belorussian, Russian, Ukrainian and other populations comparative analysis of allele frequencies was performed and Neighbour-Joining tree based on Nei distances was constructed. So, highly significant differences were observed when comparing the populations from East Europe with populations from West Europe. But although the Y-chromosome microsatellites seem to be very useful in comparing closely related populations, we have not found significant differences between Ukrainian, Russian and Belorussian populations.
For more than 360 Y chromosome assuming a stepwise mutation model haplotypes net was constructed. The three populations tested had significant differences in their haplotypes distributions. The most common haplotype (13/11/11/24/16) was found in 14% of Ukrainian individuals. In population from Russia the most common haplotype (13/11/10/25/16) was observed in 22% individuals. For population from Belorussia both of these haplotypes were most frequent - 13% and 10% respectively. Also it is interested to note that these haplotypes in Western European populations are to be found with very low frequency. On the other hand, the frequencies of these haplotypes in some Central Asian populations reaches extremely high level - up to 50%. . Perhaps, male-lineage diversity in Slavic populations underdone of migration influence from East.

I. Barisic¹, I. Ligutic¹, R. Gjergja¹, Z. Beer², Z. Modrusan-Mozetic³, A. Zuzek⁴, N. Vondracek⁵;
¹Childrens University Hospital Zagreb, Zagreb, CROATIA, ²General Hospital Varazdin, Varazdin, CROATIA, ³Clinical Hospital Centre Rijeka, Rijeka, CROATIA, ⁴Medical Centre Pula, Pula, CROATIA, ⁵General Hospital Koprivnica, Koprivnica, CROATIA.

BACKGROUND: Birth defects are the most important cause of perinatal mortality in European countries. International standardised networks of registries of patients such as the Eurocat allow data comparison and give the possibility of gaining experience in surveillance and prevention of rare diseases. AIM: To to determine population-based prevalence rates, types and epidemiological characteristics of the major congenital anomalies in northwestern Croatia using standardised Eurocat methodology. RESULTS: During the 1990-1999 period, 1274 children with congenital anomalies per 64.364 births were registered, with the mean prevalence of 19.7/10 000 births. Stability in the overall prevalence of malformations was observed, but there were differences in congenital anomaly rates between different regions (p<0.01). Limb anomalies (32.3%) and congenital heart defects (24.2%) represent the largest groups of anomalies detected, followed by the defects of the urogenital system, gastrointestinal system and central nervous system. An unusually high rate of polydactyly was observed in Varazdin (16.9/10000 compared with mean rate of 8.0/10000 for EUROCAT registries). The most frequent congenital heart defects were ventricular septal defect (14.5/ 10 000), atrial septal defect (8.4/10 000) and transposition of the great vessels (2.2/10 000). Down syndrome was the most frequent chromosomal aberration (10.7/ 10.000 births). CONCLUSIONS: Differences in congenital anomaly rates between different regions of Croatia require further investigation in order to determine whether they represent a true difference in the prevalence rates or they are due to small number variation, differences in clinical reporting or characteristics of the population (e.g. size, maternal age).

Allelic associations of COL1A1 and VDR3 genes with one-year rates of bone mineral mass loss in postmenopausal women.

M. V. Moskalenko, M. V. Aseev, I. E. Zazerskaya, V. S. Baranov;
Ott's Institute of Obstetrics and Gynecology RAMS, St-Petersburg, RUSSIAN FEDERATION.

Genetic factors play an important role in the pathogenesis of osteoporosis but the genes that determine susceptibility to pathological BMD are still not fully determined.
The alleles rates of COL1A1 and VDR genes in North-West Russian population and in the group of the postmenopausal women were investigated. The control group included 174 women without any clinical or laboratory traits of osteoporosis. The group of patients comprised 119 postmenopausal women (all of them had been in menopause for 4-5 years). Respective of BMD loss speed. two groups of the postmenopausal women were allocated after 12 months survey: women with fast loss of BMD (T-criteria decrease to 10±6,38%) and women with slow loss of BMD (T-criteria decrease during 12 month (2±8,23%). The polymorphisms of COL1A1 and VDR genes were studied by PCR-RFLP method (polymorphism COL1A1/Apa I and VDR3/Taq I).
The differences in allele combinations of VDR and COL1A1 genes between control group and the group of postmenopausal women with osteoporosis were proved. Frequencies of functionally impaired allele of COL1A1 gene (s allele) was 17,9% (control group), 2.4% (patients with slow loss of BMD), 30.8% (patients with fast loss of BMD). Frequencies of functionally impaired allele (t -allele )of VDR gene was 32,7% (control), 13.9% (patients with slow loss of BMD), 46.8% (patients with fast loss of BMD) . Significant association of BMD loss values with functionally inferior alleles for COL1A1(s) and VDR (t) genes has been proved (p<0,001).

A. V. Kirichenko;
Institute of Cytology and Genetics, Novosibirsk, RUSSIAN FEDERATION.

Modern human genetics data includes large (hundreds of people) pedigrees coming, in particular, from isolated populations. It is a problem to draw these pedigrees and to analyze the pictures due to their complex structure. Our software PedigreeQuery is based on step-by-step drawing in which a nuclear pedigree is added at each step. The direction of a pedigree extension is indicated by the user. The software helps visualizing the transmission of rare alleles, extracting the fragment of complex pedigree with identical mitochondrial genome or identical Y-chromosome.
The software uses a data file with LINKAGE format and creates PostScript files for printing. PedigreeQuery draws complex pedigrees with multiple loops, inter-generation mating, individuals with multiple mates. There are several additional features for visualization of genetic data: different colors of symbols according to the different phenotypes, crossing symbols, text underneath the person’s symbol which includes any desired information (genotype markers, name, age, etc.). Drawing pedigrees is highly configurable and may be specified by nine parameters, i.e., font and symbol’s size, space between symbols, and others.
This software can be widely used both in population genetics and genetic epidemiology.

Genetic polymorphism in Cumanian population determined by analysis of ancient bone samples

E. Szabo¹, T. Kalmar¹, F. Horvath², I. Rasko¹;
¹Institute of Genetics, Biological Research Center of Hungarian Academy of Sciences, Szeged, HUNGARY, ²Mora Ferenc Museum of Szeged, Szeged, HUNGARY.

Between 1975 and 1999 thirty-nine tombs were discovered in Csengele (Hungary). The archeologists presumed that the buried persons belonged to the Cumanian ethnic group who, according to the historical data, settled down in that area of the early Hungary during the XII-XIII centuries. The anthropological analyses of the Cumanians suggest that they might be originated from Asia, however their genetic origin has not been known.
DNA was extracted from 11 bones for studying the genetic origin of Cumanians. From the extracted DNA gender determination and mitochondrial D-loop sequence analysis has been carried out. Using X and Y chromosome specific alphoid satellite markers the gender of the 6 bone samples out of 11 were determined. Our analysis confirmed the anthropological sex identification of four adult individuals as males and was successful in cases of the female juvenile individuals. The remaining five samples did not preserve the investigated genomic DNA markers in detectable form.
The nucleotide sequences of the mitochondrial hypervariable region I were determined. The genetic homogeneity was investigated within the studied Cumanians, and the phylogenetic relationship among the Cumanian samples and modern populations was also established. One part of the investigated Cumanian bone samples carries common maternal lineages with the Asian Mongol and Buryat populations, while the other part shows common mitochondrial haplotypes with some modern European populations. We postulated that the Cumanian population originated from that area (putative homeland) where the modern Buryat and Mongol populations live, at that time when they reached Hungary they were not genetically homogenous.

V. Havasi¹, K. Komlosi¹, J. Bene¹, M. Ghosh¹, A. Nagy², K. Méhes¹, G. Kosztolányi¹, B. Melegh¹, Z. Szolnoki³, G. Melegh⁴, G. Tóth¹;
¹Department of Medical Genetics and Child Development, University of Pécs, Pécs, HUNGARY, ²1st Department of Internal Medicine, University of Pécs, Pécs, HUNGARY, ³Pándy Kálmán County Hospital, Gyula, HUNGARY, ⁴Semmelweis Hospital, Miskolc, HUNGARY.

The presence of mutations of the factor V gene such as Arg306Thr (factor V Cambridge), Arg306Gly (factor V Hong Kong) and Arg506Gln (factor V Leiden) were tested in 1360 Hungarian thrombotic patients and in 670 controls. We examined 553 hematological patients (e.g. deep venous thrombosis, pulmonary embolism) and 807 patients with different neurovascular diseases (including stroke, ischaemic cerebral attacks, sinus thrombosis, cerebral infarctions). For the FV Cambridge and Hong Kong mutation tests a PCR-based method was developed in our laboratory. After amplification of a 252 bp fragment of the gene, it was digested with MvaI. Both the above mutations abolish the cleavage site for this enzyme. Further differentiation was performed by digestion of the original PCR product by HpaII. For the factor V Leiden mutation we found 111 heterozygotes and 6 homozygotes in the hematological group (allele frequency:11.1%), 83 heterozygotes and 5 homozygotes in the neurovascular cohort (5.76%) and 43 heterozygotes and 3 homozygotes among the controls (3.66%). The allele frequencies of the Leiden mutation showed statistically significant difference in the disease groups versus the controls. In contrast to our expectations neither the factor V Cambridge nor the factor V Hong Kong mutations could be detected in the above mentioned groups. Our findings suggest that the FV Cambridge and Hong Kong mutations are even more rare than previously supposed and cannot be significant contributors to the APC resistance related thrombophilias.

Microsatellite DNA variability in the dystrophin gene in three Asian populations

J. Miranda¹, H. L. Lim², M. H. Liew², W. L. Lee¹, P. S. Low², P. S. Lai²;
¹National Neuroscience Institute, Singapore, SINGAPORE, ²National University of Singapore, Singapore, SINGAPORE.

In order to establish the informativeness of polymorphic markers in the dystrophin gene for linkage analysis in our local ethnic groups, we analysed seven multiallelic STR loci in normal X chromosomes from Han Chinese, Malay and Asian Indian populations. The markers examined were the 3‘ flanking DYSI, intragenic STR44, STR45, STR50, STR62, STRHI, and the 5‘ flanking DYSIII microsatellites located at Xp21. Allele frequencies, heterozygosity values, and pair-wise population differentiation were calculated using the GENEPOP (version 3.1d) software. The number of alleles ranged from 4 to 15, with the highest and lowest average heterozygosity values observed at STR44 (87%) and STR62 (34%), respectively. Genetic differentiation was observed between all population pairs at STR44, STR62, and STRHI. The allele frequency distributions at all loci showed varied patterns: DYSI, STR62 and STRHI showed unimodal patterns while DYSIII was bimodal; STR45 showed complex, bimodal, and unimodal patterns while STR44 and STR50 showed complex patterns in all populations. Small to significant differences were seen between the data observed in our populations and those reported for other Asian and Caucasian populations. Genetic heterogeneity found for the microsatellite loci demonstrate that these markers are effective tools for linkage analysis for non-deletion cases of Duchenne and Becker muscular dystrophy families, as well as for anthropological studies.

G. Sole¹, L. Balaci², D. Altea², N. Olla¹, C. Spada², A. Maschio¹, E. Mocci¹, S. Doratiotto¹, M. Devoto³, A. Cao¹^,2, G. Pilia¹^,2;
¹IRTAM - CNR, Selargius - Cagliari, ITALY, ²Dipartimento di Scienze Biomediche e Biotecnologie, Università di Cagliari, Cagliari, ITALY, ³Dipartimento di Oncologia, Biologia e Genetica,Università di Genova, Genova, ITALY.

There is evidence that Linkage disequilibrium (LD), the non random association of alleles at closely linked loci, vary between populations. As the feasibility of linkage disequilibrium mapping studies are critically dependent on the extent of linkage disequilibrium, we examined the patterns of LD for 24 polymorphisms in a sample from Sardinian population. Founder haplotype sets were identified by Simwalk2 program in a sample of 100 families. Microsatellite markers came from three genomic regions - 118 Kb on chromosome 12q14, 10 Mb on chromosome 13q14.2-q21.1 and 128 Kb on chromosome on chromosome 16p12.1-p11.2. For each marker pair, the strength of association was measured by a chi-square test and the standardized multiallelic D' was calculated. A useful measure is the half-length of LD - the distance at which the average D' drops below 0.5.
As expected, LD tended to be most significant for nearby markers, but occasionally markers at 6 - 6,5 Mb apart showed p< 0.05. At distances up to 200 Kb, all the marker pairs LD but one (95%) appeared significant (p<0.05). The negative correlation between D' and physical distance was striking and significant (rs = -.625, p<0.01).
Average D' value was 0.51 at distances up to 50 Kb, but it dropped to 0.28 at distances up to 100 Kb. We are at present extending this study, genotyping other markers in the same and another genomic regions. These results, if representative of the whole genome, would suggest the feasibility of whole-genome association studies in Sardinian population.

Mutation A1555G in the 12S rRNA gene and its epidemiological importance in German, Hungarian and Polish patients

S. Kupka¹^,2, T. Tóth³, M. Wróbel⁴, W. Szyfter⁴, H. Zenner², I. Sziklai³, N. Blin¹, M. Pfister²;
¹Department of Anthropology and Human Genetics, University of Tübingen, GERMANY, ²Department of Otolaryngology, University of Tübingen, GERMANY, ³Medical and Health Science Center, Department of Otolaryngology, University of Debrecen, HUNGARY, ⁴Department of Otolaryngology, University of Medical Sciences, Poznan, POLAND.

The A1555G mutation in the 12SrRNA gene has been associated with aminoglycoside induced and nonsyndromic sensorineural hearing impairment. In this study we analyzed Hungarian, Polish and German patients with nonsyndromic severe to profound hearing impairment of unknown origin for this mutation. The frequency of the A1555G mutation in the Hungarian hearing impaired population was below 1.8 %. Three out of 125 Polish patients carrying the A1555G mutation were identified. Among German patients one carrier was found (0.7 %) revealing a homoplastic A1555G mutation, whereas no mutation was detected in control individuals with normal hearing (frequency < 0.6%). In summary the frequencies of the A1555G mutation are low in the hearing impaired as well as in the normal population in Hungary, Poland and Germany. Since the importance of this mutation and its relationship with aminoglycoside exposure is not well understood yet, patients with nonsyndromic hearing impairment should be routinely screened for this mutation to avoid aminoglycoside induced hearing impairment due to increased sensitivity of maternal relatives.

S. Atanasova¹, N. von Ahsen², T. Dimitrov³, M. Oellerich², D. Toncheva¹;
¹Medical Genetics, Medical University, Sofia, BULGARIA, ²Clinical Chemistry, Georg-August University, Göttingen, GERMANY, ³Department of Nephrology, Medical University-Sofia, BULGARIA.

NAT enzymes are involved in metabolic activation and deactivation of environmental carcinogens, such as arylamines. Several allelic variants determine the phenotype of rapid or slow acetylators. Each phenotype has been associated with different risk to certain cancers, related to carcinogenic exposure. Various environmental carcinogens are suspected to be etiological factors for the chronic nephritis and for the frequent development of uroepithelial (bladder, renal pelvis and ureter) tumours in BEN patients. There is a lack of data on NAT allelic frequencies in BEN patients and in the healthy Bulgarian population. 72 Bulgarian BEN patients and 112 healthy Bulgarians as controls were genotyped for NAT1 (C1095A, T1088A, C559T, G560A, T640G) and NAT2 (T341C, C282T). Rapid cycle PCR and melting curve analysis were used for genotyping. To increase the throughput of genotyping probes were designed for temperature multiplexing where possible. The estimated frequency of the predictive NAT2 phenotype for the rapid acetylators was 0.486 in BEN patients group versus 0.464 in the control group and for the slow acetylators 0.514 versus 0.536 respectively. The frequency of the predictive NAT1 phenotype for the homozygous normal acetylators was 0.639 in BEN patients group versus 0.580 in the control group; for the homozygous rapid acetylators 0.042 versus 0.071; for the heterozygous rapid acetylators 0.292 versus 0.339 and for the heterozygous slow acetylators 0.028 versus 0.009. Homozygous slow acetylators were not identified. Allele variants and frequencies did not significantly differ between the two groups and were similar to those reported for the Caucasian population.

Interest of haplotypic tests to detect the role of a candidate gene in multifactorial diseases

P. Verpillat¹^,2, P. Margaritte-Jeannin¹, C. Bourgain¹, F. Clerget-Darpoux¹;
¹INSERM U535, Le Kremlin Bicêtre, FRANCE, ²CHU Bichat-Claude Bernard, Paris, FRANCE.

Background: When studying the potential role of a candidate gene in a multifactorial disease, information may be not available on the functional marker, but only on two neutral markers which are in linkage disequilibrium with the functional one.
Objective: To compare the power of haplotypic tests, taking into account simultaneously the information on two neutral markers, to the power of tests realised individually on each marker.
Methods: A program was written to calculate for patients and controls the frequency of haplotypes and alleles for the two markers, according to different models for the functional polymorphism (allelic frequency, different genotype penetrances) and different values of linkage disequilibrium. Power was then calculated for the haplotypic test and the tests on each marker.
Results: In many situations, the role of the candidate gene is detected by the haplotypic test when it can not be detected by the separated study of each marker. The power increase of the haplotypic test depends on the contrast of the linkage disequilibrium between patients and controls, disequilibrium which may be sometimes of contrary sign in the two samples. This situation was observed in the study of the low density lipoprotein receptor-related protein gene in Alzheimer’s disease (Verpillat et al, 2001).
Conclusion: In some situations, the use of simultaneous information on two markers of a candidate gene, using haplotypic tests, can strongly increase the power to detect the role of this gene.

Insertion-Deletion Polymorphism of the Ace Gene in Populations of North Caucasus and of Middle Asia

T. Y. Korshunova¹, A. Z. Salimova¹, V. L. Akhmetova¹, M. A. Bermisheva¹, G. M. Berezina², I. A. Kutyev¹, R. I. Fatkhlislamova¹, G. S. Svyatova², E. K. Khusnutdinova¹;
¹Institute of Biochemistry and Genetics of the Ufa scientific centre of RAS, Ufa, RUSSIAN FEDERATION, ²Republican Centre of Health Protection of Mother and Child, Almaty, KAZAKHSTAN.

Polymorphic Alu-repeats (polymorphisms consisting of the presence/absence of an Alu element at a particular location) present a very useful markers for human population genetics and evolution studies. A well studied polymorphism in the intron 16 of the angiotensin-converting enzyme (ACE) gene due to insertion (I) or deletion (D) of a 287 bp sequence has been reported to occur producing three genotypes DD, DI and II.
We have investigated the frequency of ACE insertion/deletion (ID) polymorphism in two populations of North Caucasus : Cuban Nogays from republic Karachaevo-Cherkesia (n=102) and Caranogays from republic Dagestan (n=121) and in two populations of the Middle Asia: Kazakhs (n=168) and Uzbeks (n=105). Caucasus and Middle Asia are an interesting regions for studying relative influence of linguistic variability and geographic barriers on the genetic structure of populations.
We used PCR method. PCR product was subjected to polyacrilamid gell electrophoresis. ACE genotypes frequencies for DD, ID and II were determined 19,6%, 58,3%, 22,1% in Kazakhs and 12,4%, 73,3%, 14,3% in Uzbeks and 24,5%, 58,8%, 16,7% in Nogays and 23,9%, 67,8%, 8,3% in Caranogays. The ACE allelic frequency distribution for D and I alleles was 48,8% and 51,2% in Kazakhs, 49% and 51% Uzbeks, 53,9% and 46,1% in Nogays, 57,8% and 42,2% in Caranogays.
Results of our investigations as compare to analoges of literature data.

G. F. Korytina¹, T. Victorova¹, T. Ivaschenco², B. Baranov², E. Khusnutdinova¹;
¹Institute of Biochemistry and Genetics, Ufa, RUSSIAN FEDERATION, ²Research Institute for Obstetrics and Gynecology, St. Petersburg, RUSSIAN FEDERATION.

Cystic fibrosis (CF) is one of the most common autosomal recessive disorders. A mutations CFTR gene causes this disorder. The aim of our study was to identify the most common mutations and create a CF-screening programme that would enable complete CF mutations detection among our patients. We have screened 141 CF chromosomes for 18 previously reported mutations: delF508, 1677delTA CFTRdele2,3(21kb), 394delTT, 1154insTC, R347P, R334W, G542X, G551D, R553X, S549N, 2184insA, 2143delT, S1196X, 3737delA, 3849+10kbT C, W1282X, N1303K. We also used SSCP-analysis for exons 3, 7, 10, 11, 13, 19 and 20, to find other mutations. The results of molecular analysis revealed the following frequency of detected mutations: delF508 (34%), 394delTT (3,5%), CFTRdele2,3(21kb) (1,4%), R334W (1,4%), G542X (0,7%), 2184insA (0,7%), S1196X (0,7%), 3849+10kbT>C (1,4%), W1282X (0,7%), N1303K (1,4%). Six novel mutations are detected I488M (0,7%) Q493E (0,7%) in 10 exon, 1811+12A C (1,4%) - 11 intron, T663S (1,4%) - 13 exon, I1226R ( 0,7%) -19 exon, 4005+9A>C (0,7%) - 20 intron. And two new polymorphisms was found - 2097A C (A655A) -13 exon (0,7%), 3996G>C (V1288V) - 20 exon (2,1 %). This leading study resulted in the identification of 16 different mutations accounting for almost 50% of CF alleles.

Mitochondrial DNA diversity in the populations of Middle Asia and Northern Caucasus.

M. Bermisheva¹, A. Salimova¹, T. Korshunova¹, G. Svyatova², G. Berezina², R. Villems³, E. Khusnutdinova¹;
¹Institute of Biochemistry and Genetics, Ufa, RUSSIAN FEDERATION, ²Repablican Center of health protection of mother and child., Alma-Ata, KAZAKHSTAN, ³Dept. Evolutionary Biology Tartu University., Tartu, ESTONIA.

Hypervariable segment 1(HVS-1) sequences of mitochondrial DNA together with RFLP sites diagnostic analyzed in the populations of Middle Asia - Uzbeks, Kazakhs and population of Northen Caucasus - Nogays in order to determine the haplogroup structure. The Nogays was found to have a large portion of mtDNA belonging to haplogroups observed in West Eurasian populations. The majority of the mtDNA lineages of Nogays belong to haplogroups H (23,6%) and U (20,2.%). East-Asian-specific lineages of mtDNAs were observed in 38,2% (haplogroups M,C,Z,D,G,A,B,F). On the data of mtDNA genetic variation we demonstrated that the Nogays have slightly more similarities with South-Eastern Bashkirs, which live in the Volga-Ural region of Russia than with other populations from Caucasus. The most maternal lineages of Kazakhs and Uzbeks have the Asian mtDNA haplogroups. However, the relative frequencies of this continental fraction of the mtDNA pool vary considerably over the populations studied. The frequency of Asian haplogroups in Kazakhs reached 62,38%, in Uzbeks - 52,52%. Thus, based on the mtDNA haplogroups composition analysis, the Kazakhs and Uzbeks were found to be the closest population to the East Asian populations.

V. A. Stepanov¹, V. Puzyrev¹, T. Kivisild², V. Kharkov¹, M. Spiridonova¹, S. Rootsi², I. Khitrinskaya¹, R. Villems²;
¹Institute of Medical Genetics, Tomsk, RUSSIAN FEDERATION, ²Estonian Biocentre, Tartu, ESTONIA.

We have analyzed the distribution of Y chromosome haplogroups (HGs) in the Old World with the special attention to the population of North Eurasia. We have constructed the database containing data on Y chromosome lineage`s frequency in 300 world populations including our experimental data on 30 populations of Siberia and Central Asia and all data published in the literature. We standardized the HGs nomenclature used by different authors to the unified format (Jobling et al. (1998), with modifications). Graphic representation of HGs distribution was made using Surfer software.
The general picture of the Y lineages distribution is consistent with the hypothesis of recent African origin of modern human. Several HGs marks the following migrations of modern human to Europe, Asia, Pacific and colonization of the New World.
As to population of North Eurasia, phylogeographic analysis reveals several components of different age and origin in their modern Y lineages pool, penetrating the territory through boreal and southern migration routs. Initial colonization of this territory, probably, through West, Middle and Central Asia in upper Paleolithic brings to the territory HG2 and 4. A next Paleolithic migrations from the West is traced by HG1. After last glacial maximum, mongoloid populations from the South, descendants of “austratic population” which settled the South-East Asia, penetrates the North Eurasia. The genetic traces of these migrants in male gene pool are, probably, HG10 and 26. Population migrations on the territory of North Eurasia in the Neolithic form the distribution picture of HGs 3, 9, 12 and 16.

Incidance of Genetic Thrombophilia in Turkish Women with Obstetric Complications-Preliminary findings

R. Bircan¹, C. Erzik¹, B. Saygi Erzik², F. Akbas², I. Güney¹, E. Oral², B. Cirakoglu¹;
¹Marmara University, Faculty of Medicine, Dept of Medical Biol & Genetics, Istanbul, TURKEY, ²Istanbul University, Faculty of Medicine, Dept of Obstetrics & Gynecology, Istanbul, TURKEY.

Background: Venous thrombosis is related with some mutations like Factor V Leiden (FVL) and prothrombin 20210A. Obstetrical complications such as preeeclampsia, IUFGR and some others are known to be associated with placental thrombosis. This study investigates the relation between certain gene mutations and obstetrical complications in Turkish women.
Methods: 88 pregnant women with obstetrical complication and 44 control subjects were studied. FVL, prothrombin 20210A and MTHFR C677T mutations were screened using PCR-RFLP method.
Results: Results have yet been obtained for 54 patients and 44 controls. The incidance of FVL heterozygosity was found to be 20.37% among patients, being most common in preeclamptics (7/34) followed by stillbirth (2/9) and IUFGR (2/10) groups. Only 2 stillbirth cases of 54 patients (3.7%) carry both the prothrombin 20210A and FVL mutations. Of 22 patients screened for MTHFR C677T mutation, 12 (55%) were found to be heterozygous carriers whereas the ratio of homozygotes is 9%. (2/22, both preeclamptics). In the control group, FVL mutation was found in 6.82% of the cases although prothrombin 20210A mutation was absent.
Conclusions: The study being still progressing, FVL gene mutation seems to be relatively common in pregnancies with preeclampsia than the uncomplicated ones. Both prothrombin 20210A and MTHFR C677T mutations were detected in certain groups, but the number of subjects is not sufficient for reasonable statistical analysis. However it looks likely that the suggestion coming out of some other studies to screen women with complicated pregnancies for thrombophilia markers can be supported.

Unusual frequency of the GSTM1 0/0 genotype in a Russian population of Ural region

G. V. Pavlov, G. A. Tsaur, S. V. Tsvirenko;
Ural State Medical Academy, Ekaterinburg, RUSSIAN FEDERATION.

Glutathione S-transferase M1 (GSTM1) belongs to the GST gene family. GSTM1 enzyme is responsible for the detoxification of active metabolites many potential carcinogens like trans-stilbene oxide, benzopyrene and other polycyclic aromatic hydrocarbons (PAH). Therefore GSTM1 may be important in modulating susceptibility to cancers. GSTM 1 exert part of glutathione peroxidase activity and have an important function in intracellular binding and transport of a wide variety of endogenous (steroid hormones) and exogenous (drugs) compounds. GSTM1 is polymorphic, and the null alleles result in a lack of corresponding enzyme activities. The presence of the GSTM1 null genotype is responsible for the highest level and significant induction of chromosomal aberrations.
The GSTM1 genotype was analysed from blood spots by means of PCR assay. Mutation in GSTM1 gene was investigated in a random sample of healthy subjects from villages of Ural's region (around Ekaterinburg-city), Russia
All the results were divided into 2 groups: GSTM1-positive (included GSTM1 1/1 and 1/0 genotypes) and GSTM1-negative (GSTM1 0/0 genotype). Frequency of the GSTM1 0/0 genotype in our study was more than 60%. This unusually high proportion of GSTM1-null genotype among healthy individuals could be attributed to the possible founder effect in villages which have stable populations. Also Ural region is the place where mix Caucasian and Asian populations On the other hand, It might reflect real geographical-dependent differences among Slavic populations.

D. A. Verbenko¹, T. V. Kekeeva¹, T. V. Pogoda¹, V. A. Spitsyn², E. K. Khusnutdinova³, A. I. Mikulich⁴, L. V. Bets⁵, N. A. Bebyakova⁶, V. P. Ivanov⁷, N. G. Abolmasov⁸, S. A. Limborska¹;
¹Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, RUSSIAN FEDERATION, ²Research Centre of Medical Genetics, Russian Academy of Medical Sciences, Moscow, RUSSIAN FEDERATION, ³Institute of Biochemistry and Genetics, Ufa Scientific Centre of Russian Academy of Sciences, Ufa, RUSSIAN FEDERATION, ⁴Institute of Arts, Ethnography and Folklore, National Academy of Sciences of Belarus, Minsk, BELARUS, ⁵Moscow State University, Moscow, RUSSIAN FEDERATION, ⁶North State Medical University, Arkhangelsk, RUSSIAN FEDERATION, ⁷Kursk State Medical University, Kursk, RUSSIAN FEDERATION, ⁸Smolensk State Medical University, Smolensk, RUSSIAN FEDERATION.

Most of the populations of Eastern Europe inhabit an area of long time relations between Caucasoid and Mongoloid peoples. This makes it possible to analyse the interaction during ethnogenesis of European peoples. The normal variability of minisatellite loci D1S80 (pMCT118) and 3’ApoB have been analysed in some East European populations. There are both Caucasoid populations (Belorussians, Russians) and admixture populations with different levels of a Mongoloid component (Komis, Mari, Bashkirs). Kalmyk population, belongs to Mongoloid group, was also considered. More than 1200 native individuals from 14 population samples were studied.
The analysis of minisatellite polymorphisms was carried out using the PCR and subsequent electrophoresis followed by silver staining. We detected 27 alleles of the D1S80 locus and 24 alleles of the 3’ApoB locus. Observed allele frequency distributions in Russians, Belorussians, and Mari were appeared to be similar with European populations ones. Minisatellite allele frequency distributions in Kalmyks are very special but more similar with distributions in other Asian peoples.
Genetic diversity analysis based on calculation of Nei’s genetic distances with allocation of populations studied in multidimensional space. The plots obtained revealed certain differentiation between East European ethnic groups. In spite of the patterns similarity, the total genetic variability of the 3’ApoB locus changes more extremely in comparison to total variability of the D1S80 locus. The genetic diversity of Komis and Bashkirs indicate a number of peculiarities that supposed to reflect an intermediate position of these ethnic groups between Caucasoid and Mongoloid peoples.

B. A. Malyarchuk¹, T. Grzybowski², M. Derenko¹, J. Czarny², D. Miscicka-Sliwka²;
¹Institute of Biological Problems of the North, Magadan, RUSSIAN FEDERATION, ²The Ludwik Rydygier Medical University in Bydgoszcz, Forensic Medicine Institute, Bydgoszcz, POLAND.

Mitochondrial DNA (mtDNA) control region sequences were determined in Poles (n = 436) and Russians (n = 201). Despite the high mtDNA diversity, both populations are characterized by similar pattern of mtDNA haplogroup distribution, which is also typical for many European populations studied. The analysis of mtDNA haplotype distribution has shown that both Slavonic populations share them mainly with Germans and Finns. The following numbers of the rare shared haplotypes and subclusters were found between populations analyzed: 10% between Poles and Germans, 7.4% between Poles and Russians, and 4.5% between Russians and Germans. A novel subcluster U4-310, defined by mutation at nucleotide position 310 in HVS II, was found predominantly in common between Poles and Russians (at frequency of 2%). Given the relatively high frequency and diversity of this marker among Poles and its low frequency in the neighbouring German and Finnish populations, we suggest a central European origin of U4-310, following by subsequent dispersal of this mtDNA subgroup in eastern European populations during the Slavonic migrations in early Middle Ages. This work was supported by the Russian Fund for Basic Research (00-06-80448) and by grant from the Ludwik Rydygier Medical University in Bydgoszcz (BW66/2002).

Population structure of Adygs, as revealed using DNA data and surname frequencies.

E. Balanovska¹, E. Pocheschkhova², D. Verbenko³, O. Balanovsky¹, Y. Seryogin¹, D. Subbota¹, K. Mamgetov², S. Limborska³, R. Villems⁴;
¹Research Centre for Medical Genetics, Moscow, RUSSIAN FEDERATION, ²Kuban Medical Academy, Maikop, RUSSIAN FEDERATION, ³Institute of Molecular Genetics, Moscow, RUSSIAN FEDERATION, ⁴Estonian Biocentre, Tartu, ESTONIA.

Adygs people represents indigenous population of Caucasus and belongs to the North Caucasus linguistic family. The ethnos is strongly subdivided into four tribes, each consists of some smaller groups. The most peculiar tribe is Shapsugs, which is divided from other Adygs by main Caucasus mountains chain. Analysis of inter- and intra group genetic diversity has been performed in the hierarhical population system: ethnos - tribes - villages.
Surname frequencies were obtained almost totally for all Adygean local populations (61 villages, near 60000 individuals). Nuclear DNA markers (ApoB and D1S80 minisatellites) and mtDNA markers (HVR1) have been analysed. More than 400 individuals have been studied in total.
We detected more than 16 alleles for each minisatellite loci. The geterozigosyty values are: 0.72 (D1S80) and 0.85 (3’ApoB) in Adygs; in Shapsugs 0.76 and 0.67 accordingly. As for HVR1 mtDNA, in Shapsugs 64 mitotypes and in other Adygs 80 mitotypes have been revealed (sample sizes are 107 and 239).
Fst level of intergroup diversity in whole population system of Adygs as revealed by surname data amount to 0.0209. Different tribes appeared to be quite different by Fst levels: level varies from 0.0025 for Kabardinians to 0.0253 for Shapsugs. Since, strong genetic heterogeneity has been revealed in population system of Adygs. The work was supported by RFBR grants.

Allelic association of gene markers in the FC epsilon receptor I beta gene and IL-4 gene promoter in Italian atopic children.

D. C. Salpietro¹, L. Rigoli¹, C. Di Bella¹, G. Calabrò¹, V. Procopio¹, E. Cavallaro², S. Briuglia¹, M. V. Merlino¹;
¹Unità Operativa di Genetica ed Immunologia Pediatrica Policlinico Universitario, Messina, ITALY, ²Unità Operativa di Terapia Subintensiva e Dialisi Policlinico Universitario, Messina, ITALY.

It is known the correlation between the FC epsilon receptor I beta(FCERIB) gene and IgE levels.Indeed,the beta-chain of the high-affinity receptor for IgE is found on mast cells and basophils.Crosslinking of this receptor leads to increased IL-4 production by these cells.Mutations in the FCERIB gene could alter IL-4 production and thus modify IgE levels.IL-4 is essential for B cells switching to IgE antibody production and for maturation of T-helper cells to the Th2 phenotype.A panel of 100 children(58 girls and 42 boys) with atopy was selected from 24 families of Sicily (Southern Italy).A group of 103(50 girls and 53 boys) nonatopic controls included outpatients with no history of allergic diseases was recruited.Atopy was defined by the presence at least of two of following criteria:1)a positive skin prick test;2)elevated specific IgE at least class 2;3)elevated circulating total IgE.In both groups,we have performed the molecular screening of FCERIB gene GLU237Gly and IL-4 promoter C-590T polymorphisms(RFLPs)by allele-specific PCR and PCR-SSCP.Significant difference was observed in the genotype frequency at codon 237 of the FCERIB gene between atopic children and not atopic controls (p=0.044).This RFLP was also associated to elevated levels of IgE,positive prick and rast tests.There was no difference in genotype frequency of IL-4 promoter -590C/T between the two groups.No associations were observed between this RFLP and total IgE and allergic tests.The results of our study suggests that genetic markers of atopy can be identified and could be used in the clinical setting to identify individuals at risk for atopy.

Does accounting for gene-environment (GxE) interaction increase the power to detect the main effect of a gene in a multifactorial disease ?

H. Selinger-Leneman¹, E. Genin¹, J. M. Norris², M. Khlat³;
¹INSERM, U535, Le Kremlin Bicêtre, FRANCE, ²University of Colorado Health Sciences Center, Denver, CO, ³INED, Paris, FRANCE.

Despite tremendous efforts, few genes involved in the susceptibility for complex disorders have been identified. One explanation is that these disorders are a result of an interaction between genes and environment, and under such conditions, it may be difficult to measure the true genetic effect without accounting for the interaction. Umbach and Weinberg (2000) have proposed an association test which looks at the joint effects of genotype and environment using case-parent trios. In this study, we explore under which conditions accounting for GxE interaction enhances one?s ability to detect the main effect of the gene. Using asymptotic power calculations, we investigate the power to detect the gene effect over varying exposure frequencies and several models of GxE interaction. We show that, for a given sample size, interaction model and allele frequency, the gain in power while accounting for the interaction depends on the exposure frequency : the largest gain are seen for the smallest exposure frequencies, and a loss of power can even be observed when the exposure is frequent and/or the exposure effect is strong. If we consider a gene with a disease allele frequency of 0.2, with no effect in the absence of exposure, an exposure with a 10-fold increase risk and a GxE relative risk of 2, then : when the exposure frequency is 0.1, accounting for GxE interaction increases the power to detect the gene effect in 200 trios by 10%; when the exposure frequency is 0.9 it decreases the power by 15%.

A. Milic¹, G. Piluso², V. Ventriglia², F. DAmico², B. Kovac³, A. Trlaja⁴, Z. Mitrovic⁵, N. Zurak⁵, L. Politano², N. Canki-Klain¹;
¹Croatian Institute for Brain Research and Dept of Neurology, Zagreb University Medical School, Zagreb, CROATIA, ²Dept of Pathology and Dept of Internal Medicine, 2nd University of Napoli, Napoli, ITALY, ³Dept of Neurology, University of Osijek, Osijek, CROATIA, ⁴Dept of Transfusiology, Universita of Split, Split, CROATIA, ⁵Dept of Neurology, Zagreb University Medical School, Zagreb, CROATIA.

Background. Our previous results have shown that the most frequent mutation in CANP3 gene in patients from Croatia is the 550delA mutation, while the Y537X mutation was found only in 1 family. We report the results of the screening of CANP3 gene on 27 families, 8 of them never investigated, in which one or both mutations have been identified.
Objective. To determine mutation spectrum of CAPN3 in patients from Croatia.
Patients and Methods. During a 3-year-long project concerning etiology and epidemiology of muscular dystrophies in our country, 37 patients from 27 families with potential calpainopathy were selected by clinical and family study. In the only sporadic patient the diagnosis was confirmed by CAPN3 Western blot. Beside the 550delA mutation in exon 4, two new mutations (R49H and R541W) and one gross deletion (F200-L204del) were identified by DHPLC, Transgenomic Wave System. Furthermore, we developed screening methods for these mutations which included PCR and use of restriction enzymes.
Results. Analysis of 54 CANP3 chromosomes by 5 mutations revealed the presence of 550delA in 36/54 (67%), R541W in 3/54 (5.5%), R49H in 1/54 (1.85%), Y537 in 1/54(1.85%) and delFWSAL in 1/54 (1.85%).
Conclusions. CANP3 gene screening by 5 mutations is able to identify 77% of patients with calpainopathy (LGMD2A) in our population. R541W and R49H are novel mutations.

Lack of association between endothelial nitric oxide synthase gene polymorphism and coronary artery disease in the Greek population

K. Lamnissou¹, M. Vasilakou¹, C. Maratheftis¹, C. Kasparian², N. J. Pantazopoulos², C. Deltas³;
¹Dept of Biology, University of Athens, Athens, GREECE, ²Dept of Cardiology, "Laiko" Hospital, Athens, GREECE, ³Dept of Molecular Genetics, The Cyprus Institute of Neurology & Genetics, Nicosia, CYPRUS.

Genetic polymorphism in the gene for endothelial nitric oxide synthase (ecNOS) has been considered as a potential risk factor for the development of coronary artery disease (CAD) in some populations. We studied a 27 base-pair tandem repeat polymorphism in intron 4 ecNOS gene in 105 control and 82 patients of Greek population. The patient group consisted of subjects aged under 58 years presenting with symptomatic CAD, documented by coronary angiography. The dada between the two groups were analysed by chi- square test. We found no significant difference in the frequency of 4ab genotypes between patients and controls. The frequencies for ecNOS4bb, ecNOS4ab and ecNOS4aa genotypes were 0.69, 0.28, 0.03, respectively, in controls compared to 0.71, 0.24, 0.05 in patients. Thus, in contrast to earlier findings by others from some Asian populations, we have found no evidence for an association between ecNOS4a allele as well as ecNOS4aa genotype and the risk of premature coronary disease in the Greek population.

V. Karcagi¹, I. Tourrnev², C. Schmidt³, A. Herczegfalvi⁴, V. Guergueltcheva², I. Litvinenko², I. Song³, A. Abicht³, H. Lochmüller⁵;
¹National Center for Public Health, Budapest, HUNGARY, ²Alexandroff University Hospital, Sofia, BULGARIA, ³Gene Center and Friedrich-Baur-Institute, Munich, GERMANY, ⁴Bethesda Children's Hospital, Budapest, HUNGARY, ⁵Gene Center of LMU and Friedrich-Baur-Institute, Munich, GERMANY.

Congenital myasthenic syndromes (CMS), a heterogeneous group of disorders arise from various defects of the neuromuscular transmission. The majority of CMS are inherited as autosomal recessive traits due to loss-of-function mutations of the acetylcholin receptor (AChR) subunit genes or the collagenic tail subunit of acetylcholinesterase (ColQ) gene. Many of the e subunit deficiencies, as most frequent causes of CMS, were found to be due to “private” mutations and produce clinical phenotypes with great variations.
This study summarizes data of the genetic analysis of a common mutation, a homozygous basepair deletion in exon 12 of the AChR e subunit gene (e1267delG) in 66 CMS patients from 46 non-related families. All patients were clinically characterized as having sporadic or autosomal recessive CMS. All e1267delG families were of Romani (Gypsy) and/or southeastern European origin. Phenotype analyses revealed a uniform pattern of clinical features including bilateral ptosis, moderate fatigable weakness of ocular, facial, bulbar and limb muscles, positive response to anticholinesterase treatment and a benign natural course of the disease. Genotype analysis was carried out and indicated a common ancestor (founder). We conclude that the mutation e1267delG might be the most frequent cause of CMS in patients of Romani (Gypsy) ethnic origin. Therefore, carrier and/or newborn testing may be beneficial in this group, as the disease is treatable. Moreover, additional haplotype studies in patients harboring e1267delG may enable a more accurate age-dating of this particular mutation. This may help to understand the origin, evolution and dispersion of the disease in the Romani populations.

S. M. Echwald¹, L. H. Larsen¹, T. I. A. Sørensen², T. Hansen¹, T. Andersen³, O. Pedersen¹;
¹Steno Diabetes Center, Copenhagen, DENMARK, ²Danish Epidemiology Science Centre at Institute of Preventive Medicine, Copenhagen, DENMARK, ³Roskilde County Hospital, Roskilde, DENMARK.

Background: Human obesity is a prevalent, complex disorder, which is now reaching epidemic proportions in the Western world. It is assumed that genetic disposition to obesity determine susceptibility to environmental factors. Discovery of the molecular basis for obesity in the Agouti mouse has implicated the melanocortin 4 receptor (MC4R) in the development of obesity in rodents as well as in man. Aim: The aim of the present study was to analyze the prevalence of MC4R mutations in a Danish sample of obese subjects. Subjects and Methods: Using PCR-dHPLC and direct sequencing, we screened a cohort of 751 obese male subjects with early-onset obesity recruited at the draft board examinations as having a BMI ³ 31 kg/m² (BMI=33,3±2,4 kg/m²) and a control group consisting of 706 randomly sampled draftees (BMI=21,4±2,1kg/m²).
Results: A total of 14 different nucleotide substitutions were identified. Among these, 11 nucleotide substitutions were found only in the obese study cohort. Among these, 9 carriers of a Tyr35Ter variant and 6 carriers of the Arg165Gln variants were identified and showed direct association to obesity (p= 0.004 and p=0.02, respectively). Combined, the carrier prevalence of mutations in the MC4R among obese Danish men is 2.8%. In conclusion, we find a 2.8% carrier frequency of possible functional variants of the MC4R gene among obese Danish subjects which is consistent with reported carrier frequencies in other European populations, making the genetic variability in the MC4R the most common cause of genetically determined human obesity with known aetiology.

The Kingdom of Saudi Arabia occupies over 2/3rd of the Arabian Peninsula The overwhelming majority of the population of Saudi Arabia are Arabs belonging to different nomadic tribes. In early 1960s, investigations began to identify the nature of genetic disorders in the Saudi population and to determine the frequency of the common abnormal genes. Among the single gene disorders, a large number of disorders are identified and some exhibit polymorphism in certain regions of Saudi Arabia. These include sickle cell, a- and b-thalassaemia and glucose-6-phosphate dehydrogenase deficiency genes. A significant number of amino acidurias and other inborn errors of metabolism have been reported. The majority of these disorders contribute significantly to the development of mental retardation in Saudi population. Cystic fibrosis, congenital hypothyroidism, Bardet-Biedl Syndrome, fragile X syndrome, Lower syndrome, and a number of other genetic disorders, have been reported amongst the Saudis. Chromosomal disorders contribute significantly to morbidity and mortality in the Saudi population. Amongst the most frequent Down’s syndrome which occurs at a frequency of 1 in 800. Multifactorial disorders are the most common genetic defects and are significant factors underlying morbidity amongst the Saudis. The community studies clarified several of these disorders including diabetes mellitus, hypertension, obesity and cardiovascular diseases. A comprehensive national study has determined the contribution of genetic causes in the development of disability in different regions of Saudi Arabia.
This paper will present an overall coverage of genetic disorders in the Saudi population and suggest preventive measures.

Genetics diversity of microsatellite loci in populations of Siberia and Central Asia

M. Spiridonova, V. Stepanov, V. Puzyrev;
Institute of Medical Genetics, Tomsk, RUSSIAN FEDERATION.

1600 individuals from 23 populations belonging to 13 ethnic groups living on the territory Siberia and Central Asia was investigated. These are Northern and Southern Altai, Tuvinian, Buryat, Evenk, Yakut, Kirghize, Uzbek, Tajik, Dungan, Russian, Ukrainian, Tatar. Nine autosomal (D4S397, D5S393, D7S640, D8S514, D9S161, D10S197, D11S1358, D12S364 and D13S173) microsatellites loci (STR) have been studied. Automated genotyping of STR loci was performed with HEX-, TET- or FAM-labeled primers with the ABIPrism310 genetic analyzer and Genescan software. High level of microsatellite loci diversity was shown. In total, 128 different allels at nine studied loci were found. Only 2.5 % of genetic variability of microsatellites was attributable to the differences between populations, whereas 97.5% of genetic diversity was found within populations. Autosomal STR system is most adequately for detection of genetic relationship between closely-related group of populations. The greatest level genetic differentiated of local populations was observed in three Tuva populations (FST=2.68%). Principal component analysis and phylogenetic analysis showed substantial difference between gene pool of Altai and Indo-European populations. High similarity of gene pool in local populations within certain ethnic group was demonstrated both by principal component and phylogenetic analysis. Phylogenetic analysis revealed genetic close relation between Evenks and Buryats.

Demogenetic study of three populations within a region with strong founder effect

M. Tremblay, È. Lavoie, L. Houde, H. Vézina;
University of Quebec at Chicoutimi, Chicoutimi, PQ, CANADA.

For more than three decades, the Saguenay population (Quebec, Canada) has been the subject of many studies in population genetics. It has been shown that 82% of the region's gene pool originated from nearly 2600 founders who settled in New-France during the 17th century. Some of these founders are believed to have introduced rare deleterious genes in the Quebec population. The demographic behaviour of these founders' descendants is at the heart of the process by which some of these rare alleles were transmitted and are now found at an elevated frequency in the contemporary Saguenay population. This study aims to a better understanding of the origins and stratification of the Saguenay gene pool. The region has been divided in three sub-populations based on geographical and historical criteria: Lower Saguenay, Upper Saguenay and Lac St. Jean. Three hundred extended genealogies (100 for each sub-population) have been reconstructed, using the BALSAC population register. These genealogies have an average depth of 10 generations. Kinship and inbreeding measurements, as well as the founders' genetic contribution, frequency and number of occurrences were examined. Founders' geographical origins, intraregional migratory movements as well as frequency and concentration of surnames were also considered. Preliminary results indicate lower inbreeding coefficients within the Upper Saguenay population, which is more industrialized and has benefited from a dynamic migratory input. Kinship coefficients between the three sub-populations become distinguishable from the 6th generation onward; highest values are those between Lower Saguenay and Upper Saguenay.

T. V. Kosyankova, N. R. Maksimova, A. N. Kucher, V. P. Puzyrev, S. V. Buikin;
Institute of Medical Genetics, Tomsk, RUSSIAN FEDERATION.

The study of three polymorphism's (G894T, C774T and VNTR) of endothelial nitric oxide syntase gene (NOS3) in Siberian populations of different race and ethnic specificity (Russians, Tuvinians, Buryats, Yakuts) was carried out. For the studied markers the distributions of genotypes in the most populations corresponded to Hardy-Weinberg equilibrium (HWE). A deviation from HWE was shown only for C774T in Russian and Tuvinians (p<0.01). There were an excess of allele C774 (82%) in Russian and G894 (69%) and VNTR B (93%) in Buryats. Have been shown significant difference in alleles prevalence for all markers between Russian and Buryats and for VNTR between Russian and Tuvinians.
These data append accumulating information on Siberian populations gene-poll; and there are all reasons to deem that the studies in this field will help to resolve both fundamental biological issues, and appliedones, medical, that is especially actual in the light of genomic medicine development.

Low density lipoprotein receptor gene mutations and polymorphisms in St.-Petersburg patients with familial hypercholesterolemia.

F. M. Zakharova, V. V. Egorov;
Institute for Experimental Medicine, St.-Peterburg, RUSSIAN FEDERATION.

Familal hypercholesterolemia (FH) is one of the most common inherited diseases (1:500), leading to atherosclerosis and premature myocardial infarctions. FH is caused by mutations in the human low density lipoprotein (LDL) receptor gene. The early diagnosis of disease by means of DNA analysis allows to perform genetic counseling and to prevent the development of the disease in the FH patients by drug treatment. Aiming to identify the FH-causing mutations of the LDL receptor gene we have screened the separate exons of the gene in collection of 42 FH probands by means of PCR/SSCP analysis followed by DNA sequencing. During this study we have identified the following DNA mutations: C74X (c.285 C>A, TGC->TGA), E397X (c.1252 G>T, GAG->TAG), G571E (c.1775 G>A, GGG->GAG), that seems to be causative for FH development, and two neutral mutations: H229H (c.750 T>C, CAT->CAC), T705I (c.2177 C>T, ACC->ACT). All the mutations with the exception of C74X and T705I were new and not reported elsewhere. Besides the mutations we have identified five neutral changes considered as DNA polymorphisms, namely c.66 C/T (C18C, TAC->TAT), c.1171 G/A (A370T, GCC->ACC), c.1773 T/C (N570N, AAT->AAC), c.1959 C/T (V632V, GTC->GTT), c.2231 G/A (R730R, CGG->CGA). We have compared the occurrence of the polymorphism alleles in FH group and in the group of control (68 non-related subjects) that have not differed besides allele C of c.66 C/T polymorphism and allele A of c.1171 G/A polymorphism were more common in control rather than in FH group.

Impact of Misspecifying Parental Relationships in Maximum Lod Score Affected Sib-Pair Method

A. L. Leutenegger¹^,2, E. Génin¹, E. Thompson², F. Clerget-Darpoux¹;
¹I.N.S.E.R.M. U535, Le Kremlin-Bicetre, FRANCE, ²University of Washington, Seattle, WA.

Many linkage studies are done in small isolated populations and populations where marriages between relatives are encouraged. In these populations, missing information on pedigree structure is quite frequent. Here, we study the impact of misspecifying the parental relationships of the sib-pairs in the Maximum Lod Score method (Risch, 1990) and on the triangle constraints (Holmans, 1993).
Characterising the parental relationships by the kinship coefficient between the parents (f), the maternal inbreeding coefficient (a_m) and the paternal inbreeding coefficient (a_p), we studied the behaviour of the identity by descent (IBD) vector expected under the null hypothesis of no linkage with respect to these quantities. We find that the expected IBD vector is not anymore (0.25, 0.5, 0.25) when they differ from zero. There is an increased probability of sharing one or two alleles IBD. And in some cases the vector is even outside the triangle constraints.
We simulated data on two different family structures: (1) parents are double first cousins (f = 0.125, a_m = a_p = 0), (2) each parent is offspring of first cousins (f = 0, a_m = a_p = 0.0625). We then analysed the data underestimating the parental relationships. We find that ignoring the kinship and/or inbreeding of the parents increases the type I error of the test when data on the parents are not available. But when parents are typed, we observe a decrease in type I error. We also point out some unpleasant feature of the software GENEHUNTER even when parental relationships are properly specified.

Evaluation of a cluster of congenital abnormalities particularly Down syndrome in a small Hungarian village

L. Timar¹, Z. Viragh², K. Sperling³, A. Czeizel⁴;
¹National Centre of Health Promotion, Budapest, HUNGARY, ²National Centre of Public Health, Budapest, HUNGARY, ³Inst. of Human Genetics, Berlin, GERMANY, ⁴Found. of Comm. Contr.of Hered. Dis., Budapest, HUNGARY.

A unique cluster of congenital abnormalities, particularly Down syndrome and twins was detected in a small Hungarian village in 1989-1990.
Of 15 livebirths 11 (73.3 %) were affected by congenital abnormalities and 6 (40 %) were twins. Of eleven malformed babies 4 had Down syndrome.
The usual causes of congenital abnormality clusters : familial inheritence, consanguinity, classical teratogenic factors including alcohol could be excluded.
Different approaches of field studies, including a case-control study, and laboratory examinations indicated the germinal mutagenic and teratogenic effect of the excessive use of trichlorfon at local fish farms.
The content of this chemical was very high in fish (100 mg/kg) and several pregnant women, including all mothers of babies with Down syndrome, had consumed contaminated fish in the critical period for the congenital abnormalities observed.
Here the main experiences of the environmental abuse are summarized.

Gene-environment interaction in hereditary haemochromatosis: effect of excessive alcohol use on iron overload in patients homozygous for the C282Y mutation

M. C. Mérour¹, V. Scotet², A. Y. Mercier¹, B. Chanu¹, T. Le Faou³, O. Raguénès², G. Le Gac¹, C. Mura², C. Férec¹^,2;
¹EFS-Bretagne, Brest, FRANCE, ²INSERM EMI 01-15, Laboratoire de Génétique Moléculaire, Brest, FRANCE, ³EFS-Bretagne, Quimper, FRANCE.

Hereditary haemochromatosis (HHC) is the most common inherited disorder in Caucasian populations. Characterised by an iron overload, the disease is associated with increased risk of hepatocarcinoma but is effectively treated by phlebotomies. The HFE gene was cloned in 1996 and a single mutation (C282Y) is responsible for 80-95% of cases. HHC presents a genetic heterogeneity and its expression can be modified by environmental factors. The aim of this study was to identify the influence of alcohol use on the intensity of iron overload, measured by serum ferritin, in patients homozygous for the main mutation.
We retrospectively registered patients C282Y/C282Y treated in a blood centre of western Brittany. A clinical questionnaire was completed at their first visit, informing on biological and clinical signs, life customs, …. The analysis was made using a linear regression model.
This study included 351 subjects of whom 60.4% were males. The most frequent clinical signs were: weakness (58.5%), arthritis (44.7%), melanoderma (28.1%) and hepatomegaly (14.8%). Twenty-five patients had an excessive alcohol use (>60 g/day - 7.2%). Those subjects presented a significantly increased serum ferritin (1865.5 vs 940.3 ng/ml - p<0.001) and a higher risk of hepatomegaly and melanoderma. The result of regression analysis remained unchanged after adjustment on sex and age at onset.
Excessive alcohol use accentuates iron overload in C282Y/C282Y patients. This aggravating factor increases the risk of cancer and of cirrhosis. Consequently, it is recommended to those patients to have a reasonable alcohol use. This study illustrates an interaction between genetic and environmental factors.

E. Pennarun¹^,2, E. Metspalu², T. Kivisild², H. El Amri³, S. El Kabbaj³, A. Chaventré¹, R. Villems², J. P. Moisan¹;
¹LEPA, faculte de medecine, Nantes, FRANCE, ²Estonian Biocentre and Tartu University, Tartu, ESTONIA, ³Laboratoire de Recherche et d'Analyse Medicale, Gendarmerie Royale, Rabat, MOROCCO.

Moroccan mtDNA sample (N~540) was collected from different localities of the country and an ethnic origin of individuals (~320 Arabs and ~220 Berbers) has been established for the last 100 years.The variability of the sample was studied using mtDNA hypervariable region sequencing and extensive RFLP typing of phylo-genetically informative coding region positions. Sub-Saharan African lineages (L) comprise more than a fifth of the sample, being, however, somewhat more frequent among Arabs than Berbers, though a relative proportion of subtypes of haplogroup L does not differ among the two groups. In both populations, the rest of the lineages is split between 10%-11% African-specific varieties (U6 and M1) and those, typically found in western Eurasia. Meanwhile, Berbers possess haplogroup V at three-fold higher frequency than Arabs (~6.4% and ~2.2%, respectively) while the content of the pre-V variants differs less (~4.6 and 2.8%). Furthermore, Moroccan Afroasiatic speakers differ from northeastern African and Near Eastern Arabs (as well as from Ethiopians) in one more important aspect: while the latter populations possess a significant proportion of a variety of sister branches of H and V, in particular, preHV, such mtDNA variants are much less frequent or nearly absent in Morocco, whereas haplogroup V is virtually absent in northeast Africa and in the Near East. That, and several other phylogeographic arguments suggest a limited recent migration of female lineages alongside the northern Africa and should be considered as an important aspect where the spread of languages and genes is discussed.

Maternal legacy of Bretons reveals common features with the extant Scottish mtDNA

J. P. Moisan¹, E. Pennarun¹, K. Tambets², R. Villems², A. Chaventre³;
¹Laboratoire d'Etude du Polymorphisme de l'ADN (LEPA), Faculte de Medecine, Nantes, FRANCE, ²Estonian Biocentre and Tartu University, Tartu, ESTONIA, ³Laboratoire d'Etude du Polymorphisme de l'ADN, Faculte de Medecine, Nantes, FRANCE.

Information about the variability of maternal lineages in France is very limited. Here we bring new data about 400 mtDNAs from different localities in France with a specific emphasize to Brittany. We have analyzed about 120 mtDNAs of native northern Bretons by HVR sequencing and extensive search for coding region polymorphisms. Phylogenetic analysis was carried out using median networks. All found in Bretons mtDNA lineages belong to well-established European-specific haplogroups H, I, J, K, T, U, V and W. French mtDNA samples from different localities were analyzed likewise. The obtained phylogenetic trees were compared with mtDNA variability in English, Scottish, Irish, Welsh, German and Iberian populations, known from published data. This extensive search over more than 3000 mtDNAs revealed that, apart of trivial overlaps among common all over western Europe mtDNA lineages, there is a subset of more rare mtDNA variants present both in Bretons and Scots. It may suggest that a part of maternal inheritance of northern Brittany gene pool may come from Scotland, in accordance with historical data indicating the replacement of the imperial Roman Legions from the northern England to the coastal Brittany some 1500 years ago.

H. V. Tolk¹, M. Pericic², L. Barac², S. Cvjetan², P. Rudan², K. Tambets¹, R. Villems¹;
¹Estonian Biocentre and Tartu University, Tartu, ESTONIA, ²Institute for Anthropological Research, Zagreb, CROATIA.

Mitochondrial DNA lineages of three South Slavonic-speaking populations of the northwestern Balkan peninsula - Croats, Bosnians, and Slovenians (N ~1,200; ~370 haplotypes) - were identified combining the sequences of mtDNA HVS-I region and the RFLP data from coding region. These lineages were compared with a dataset of about 12,000 samples from elswhere. This phylogeographic knowledge base was used to interpret demographic events of the past since the peopeling of Europe. An absolute majority of the lineages found belong to the common western-Eurasian haplogroups - H, I, J, K, T, U, V, and W. Low-frequency haplogroups, e.g., N1, R, HV, and pre-HV, are present as well. Lineages, characteristic for sub-Saharan Africa or eastern Eurasia, occurred in single cases. For better phylogeographic resolution the data of the populations from different geographic areas were compared in a sub-haplogroup level, and the fraction of the identical haplotypes between population groups was determined. The distribution and diversity of many subhaplogroups reveals that the gene pool of the populations of northwestern Balkans has not gained much influence from the Near East during the Holocene. With some interesting exceptions, southern Slavs tend to have more common phylogenetic branches shared with Germanic (e.g. T2), West Slavonic, or, in some cases, with Finno-Ugric speakers (e.g., U4, U5), but significantly less so with southern European and eastern Mediterranean populations.

Alu-deletion polymorphism at CD4 locus correlates with Mongoloid component in the gene pool of Northern Eurasia population.

I. Y. Khitrinskaya¹, V. Stepanov²;
¹Institute of Medical Genetics,, Tomsk, RUSSIAN FEDERATION, ²Institute of Medical Genetics, Tomsk, RUSSIAN FEDERATION.

Insertion-deletion Alu-polymorphism at the CD4 locus located on chromosome 12 has been studied in more than 1400 individuals sampled from 11 ethnic groups inhabiting Siberia and Central Asia. The investigated populations related to two basic racial types of Eurasia: Mongoloid and Caucasoid. In contrast to other polymorphic Alu-insertion the presence repeat (allele Alu+) is ancestral for insertion at the CD4 locus. Primates (chimpanzees, gibbons and gorillas) are found to be monomorphic relative to insertion presence. All studied ethnic groups are polymorphic at the CD4 locus. It is interesting to note that deletion frequency (allele Alu-) decreases with increased Mongoloid component in the population. Maximal frequency of deletion allele is shown in Russians (0.339). It decreases to the east and is minimal in Yakuts (0.025). For Caucasoids living in Central Asia, i.e. Tajiks and Uzbeks the frequency of Alu allele was 0.143 and 0.130, while those for Mongoloids, i.e. Kazakh and Kirghiz was 0.129 and 0.125, respectively. In the territory of East Siberia the intensity of Mongoloid signs increases in the following series: Tuvinians, Buryats, Dungans, Evenks and Yakuts, while the frequency of Alu-deletion decreases as 0.074; 0.051; 0.045; 0.043 and 0.025, respectively. The study proved the importance of analyzing insertion-deletion polymorphism at the CD4 gene, which is very informative. This polymorphism may probably serve as a marker correlated with the degree of the Mongoloid component in populations. It may be also useful for the specialists of related fields - ethnographers, archaeologists and anthropologists.

Association between Matrix Metalloproteinase-1 (+2506) gene polymorphism and Early Onset Periodontal Disease (EOP)

M. R. Bazrafshani¹, W. E. R. Ollier¹, M. H. Thornhill², A. Hajeer¹;
¹The Centre for Integrated Genomic Medical Research, University of Manchester, Medical School, Manchester, UNITED KINGDOM, ²Oral Disease Research Centre, St Bart's and The Royal London School of Medicine and Dentistry, London, UNITED KINGDOM.

Background: Periodontal disease is a chronic inflammatory disease of the supporting tissues of the teeth, starting with gingivae and progressing to gradual destruction of the bony support and peridontal attachment of the teeth. Early onset periodontal diseases (EOP) are a group of inflammatory disorders characterised by a rapid rate of periodontal tissue destruction, in young individuals. There is now substantial evidence to suggest that genetic factors play a role in the pathogenesis of EOP.
Polymorphisms in the MMP-1 gene, which may underpin inter individual differences in MMP-1 synthesis and secretion have been associated with other diseases, which have an inflammatory pathogenesis. Genetic variation within candidate genes in EOP patients may represent a mechanism by which individuals are rendered susceptible to disease.
Objective: To investigate whether three biallelic polymorphisms occurring within the exonic regions of the MMP-1 gene (positions +85^*, +2506^** and +2613^**) are associated with EOP.
Methods: The MMP-1 polymorphisms were detected using a PCR-SNP-SHOT method. MMP-1 polymorphisms were examined in 72 patients with EOP and 91 healthy matched UK controls.
Results: No differences in allele and genotype frequencies were observed for the +85 and +2613 SNP’s. In contrast significant increases in the +2506 A allele and A/G genotype were observed in EOP cases.


	Controls		Cases
MMP-1 (+2506)	N=91	%	N=72	%
Genotype
G/G	78	85.7	41	56.9
G/A	11	12.1	29¹	40.3
A/A	2	2.2	2	2.8
Allele
G	167	91.8	111	77.1
A	15	8.2	33²	22.9
¹O.R.=4.9, C.I. 95% 2.3-10.6, c² =17.3, P<0.05 ²O.R.=3.3, C.I. 95% 1.7-6.3, c² =13.8, P<0.05

Conclusion: Several polymorphisms exist in the MMP-1 gene that influences the MMP-1 biological activity. Our results demonstrated association for EOP risk with the A allele and G/A genotype of the MMP-1 (+2506) gene.
^*Exon 1
^**Exon 5

"Implication of population genetics dimension in unrelated bone marrow transplantation organisation"

P. A. Gourraud¹, C. Raffoux², A. Cambon-Thomsen¹;
¹INSERM UNIT 558, Toulouse, FRANCE, ²FGM, Paris, FRANCE.

The human major histocompatibility complex (MHC) which contains polymorphic multicopy genes such as HLA (Human Leukocyte Antigen), is particularly interesting in terms of polymorphism and genetic markers.
For bone marrow transplantation, most allogeneic transplants are from HLA-identical sibling donors. However many patients do not have a suitable familial donor and transplants from unrelated volunteer donors are required.
Since 1980, Donor Registries have gathered more than 7 x 10⁶ potential donors worldwide.
We analysed the 100 000 HLA- A-B-DRB1 typed French donors, and patients since 1998. Haplotype frequencies, regional distribution and comparison between donors and patients HLA distribution, provide good basis for the optimisation of new recruitment strategies. We assume that within the MHC "Polymorphic Frozen Blocks", new SNP inside and outside the genes and Microsatellites would characterise some genetic profiles (including non-HLA genes). Pre-test for such markers would help choosing which entering donors to fully HLA type.
Some of 150 highly polymorphic MHC Microsatellites are in linkage disequilibrium with HLA alleles. We tested the ability of 6 Msat to predict HLA types on 800 DNA. To investigate differences and similarities compared to SNP, we reanalysed HLA class 1 alignment. These technical and genetic issues associated with economical considerations contribute to imagine new scenarios for current BMT organisation where the most polymorphic human system is playing a central clinical role. [Work performed as part of an EU contract QLG7-CT-2001-00065: MADO].

The effect of DNA polymorphisms in angiotensin I-converting enzyme and angiotensin II type1 receptor genes with arterial blood pressure levels in Serbian population

A. Stankovic, M. Zivkovic, T. Djuric, D. Alavantic;
Institute of Nuclear Sciences "Vinca", Belgrade, YUGOSLAVIA.

Polymorphism of angiotensin I-converting enzyme gene (ACE) and angiotensin II receptor gene (ATR1), the crucial components of the renin-angiotensin system, have been investigated with respect to regulation of arterial blood pressure. Aim of the study: Possible association between polymorphisms of the I/D in the ACE gene, and the A1166C of the ATR1 gene and arterial blood pressure and serum lipid levels in human population from the Belgrade area.Subjects and Methods: We have investigated 285 healthy persons recruited from the general population. Genotyping was performed by polymerase chain reaction (PCR) using a modified tree primer method for I/D and genotyping of A1166C by using allele specific amplification.
Results: Frequencies of the genotypes were a) for ACE - 0,18 (II), 0,51 (ID), 0,31 (DD), with allele frequencies 0,43 (I), 0,57 (D); b) for ATR1 - 0,54 (AA), 0,38 (AC), 0,08 (CC) with allele frequencies 0,73 (A), 0,27 (C). In males there is a correlation of the DD genotype with hypertension (OR=4,25; p=0,05). Frequencies of genotypes and alleles of A1166C significantly differ between the hypertensive males and controls (p<0,05). The male persons with CC genotype have an three time higher relative chance of being hypertensive. In two-way ANOVA, a synergistic effect of ACE and ATR1 on the total cholesterol and LDL cholesterol was obtained. The male persons with DD/CC genotype have an eight time higher relative chance of being hypertensive. Conclusion: The results of this study are a contribution to prevention and therapy of hypertension and cardiovascular disease in our population.

A. Anichkina, S. Tverskaya, A. Polyakov;
Research Centre for Medical Genetics, Moscow, RUSSIAN FEDERATION.

Phenilketonuria (PKU), one of the common inborn error of amino acid metabolism, is an autosomal recessive disease caused by mutations in phenilalanin hydroxylase gene (PAH). Up to date it is known more than 400 mutations but in every population there are some most frequent for each other. We have create the multiplex system for ACRS PCR analysis for most prevalent mutations in Europe (R408W; R158Q; P281L; I65T; IVS10-11g->a; IVS12+1g->a; R261Q; R252W) and carried out the mutation analysis in PKU patients from Moscow region. Among 94 alleles studied 47 (50%) were positive for R408W mutation, 8 (8.7%) for IVS10-11g->a mutation, 6 (6.3%) for P281L mutation, 2 (2.2%) for R252W mutation and 1 (1%) for IVS12+1g->a. No allele was positive for R158Q, I65T and R261Q mutations. The total informative of our system was 68% for our patients. The informative of this system were 87.4% for St-Petersburg PKU patients and 80.5% for Novosibirsk PKU patients (by published date). The R158Q and R261Q substitutions were detected in this regions but the frequent of Mediterranean origin mutation IVS10-11g->a were less (0.7% and 1.5% correspondently). All alleles with R408W mutation for which it was possible to determined the gaplotype (n=10) had the same VNTR3/STR234 minigaplotype. We propose that the population from central Russia include Moscow region has the other frequent mutation(s) differ from other and more sensitive technique for its detection has to be used.

M. V. Derenko¹, T. Grzybowski², B. A. Malyarchuk¹, J. Czarny², G. A. Denisova¹, M. Wozniak², I. K. Dambueva³, C. M. Dorzhu⁴, D. Miscicka-Sliwka², I. A. Zakharov⁵;
¹Institute of Biological Problems of the North, Magadan, RUSSIAN FEDERATION, ²The Ludwik Rydygier Medical University in Bydgoszcz, Forensic Medicine Institute, Bydgoszcz, POLAND, ³Institute of General and Experimental Biology, Ulan-Ude, RUSSIAN FEDERATION, ⁴Tuva State University, Kyzyl, RUSSIAN FEDERATION, ⁵Vavilov Institute of General Genetics, Moscow, RUSSIAN FEDERATION.

Based on mitochondrial DNA and Y-chromosome variability data the genetic structure of seven Southern Siberian groups, Tuvinians, Todjins, Burjats, Sojots, Tofalars, Altaians and Khakassians (the total sample size is 480 individuals) was described. It was shown that populations studied were formed on heterogeneous genetic substratum encompassing both Asian and West-Eurasian components. Phylogeographic analysis of Europeoid lineages specific for Siberian populations confirms the "southern" origin for the majority (60%) of mtDNA lineages and assumes that their occurrence is due to migrations of Europeoids from West Asia. It was shown that the Y-chromosome lineages of HG10 as well as mtDNA lineages of five (A, B, C, D, X) haplogroups specific for Native Americans are present in Altai and Sayan populations gene pools. This fact together with the presence of palaeo-Europeoid component, represented by Y-chromosome 92R7T-lineages, in Altai and Sayan populations gives an evidence of the participation of South Siberian aboriginal groups in peopling of Americas. The high frequencies of Y-chromosome HG12, which is ancestral to HG16, in populations of Altai and Sayan region (up to 30% in Tofalars) were revealed thus indicating the autochthonous Siberian origin of Y-chromosome TAT-C lineages. The results of phylogenetic analyses based on mtDNA and Y-chromosome variability data testify the considerable inter-population differentiation of Southern and Eastern Siberian groups studied. The degree of genetic differentiation is defined rather by geographical, historical and evolutionary factors, than linguistical and anthropological ones. This work was supported by RFBR (99-06-80430) and the Ludwik Rydygier Medical University in Bydgoszcz (BW66/2002).

Nonsyndromic hereditary hearing loss in Tunisia: Molecular study and impact of consanguinity

S. Masmoudi¹, A. Ghorbel², A. Elgaied-Boulila¹, M. Hmani¹, S. Ben Arab¹, A. Kassab³, S. Hachicha⁴, M. Drira², H. Ayadi¹;
¹Laboratoire de Génétique Moléculaire Humaine, Faculté de Médecine de Sfax, Sfax, TUNISIA, ²Service d'O.R.L., C.H.U. H. Bourguiba, Sfax, TUNISIA, ³Service d'O.R.L., C.H.U. F. Bourguiba, Monastir, TUNISIA, ⁴Service d'O.R.L., C.H.U. Rabta, Tunis, TUNISIA.

In industrial countries, about 60% of congenital hearing impairment cases are due to genetics defects and about 80% of hereditary hearing loss are nonsyndromic recessive deafness. Although more than 30 recessive genes have been localised, mutations involving the Cx26 gene (DFNB1) are the most common cause of deafness in many populations. In order to determine the genetic basis and the impact of consanguinity on the hereditary nonsyndromic hearing impairment in Tunisia, we have conducted an epidemiological and molecular study using 117 families with at least two affected children and 236 healthy individuals. These families were ascertained from deaf schools from the whole of Tunisia. Clinical investigation was done on all affected and unaffected individuals. Linkage analysis was undertaken in informative families and sequencing was done when families were found to be linked to one DFNB known gene. In all families and unrelated controls, Cx26 mutations were screened by DGGE and/or sequencing. Our results indicated that all families present a recessive mode of inheritance and at least 8 different DFNB genes are involved. Homozygous mutations in the Cx26 gene was revealed in 21 (18.6%) unrelated families. The most frequent mutation found was the 35delG in patients (86.5%) and in the healthy population (1.3%). On the basis of the known apparent mean consanguineous coefficient, we have determined the increase rate (3.40) of recessive deafness due to the 35delG mutation. In conclusion, the consanguinity increases the frequency of recessive deafness gene but didn’t alter the genetic heterogeneity in Tunisia.

The presence of the western Eurasian mtDNA haplogroup U5 in sub-Saharan Africans

A. Rosa¹^,2, E. Metspalu², V. Macaulay³, A. Brehm¹;
¹Madeira University, Madeira, PORTUGAL, ²Estonian Biocenter and Tartu University, Tartu, ESTONIA, ³Dept. of Statistics, University of Oxford, Oxford, UNITED KINGDOM.

We have studied mtDNA variation in 370 people from Guinea-Bissau from different ethnic groups. More than 90% of the mtDNAs belong to the sub-Saharan African haplogroups within L1, L2 and L3. Haplogroups M1 and U6 - the two African subsets of macrohaplogroups M and U, respectively - are present at much lower frequencies and exhibit little diversity. Surprisingly, the typically western Eurasian haplogroup U5 was found at a frequency of ~3%. All of these U5 mtDNAs belong to individuals from the Fulbe subpopulation of our sample - people who inhabit sub-Saharan Africa from Sudan to Senegal. U5 appears to be among the earliest branches of mtDNA outside Africa and can be found all over western Eurasia. Since U5 is either absent or extremely rare in Sudan, Ethiopia and Egypt and rare in the Near East, it seems that its presence in the Fulbe of Guinea-Bissau is best interpreted as a result of gene flow from north-western Africa, where U5 is present both among Arabs and Berbers. In contrast, our sample lacks the most frequent west Eurasian lineage clusters, e.g. H, J and T, suggesting that a specific founder event and not recurrent maternal gene flow was operable here.

S. S. Mastana¹, S. S. Papiha², P. Singh³, M. Singh³, K. Das⁴, A. Pacynko¹, P. Fisher¹, M. Das⁴, N. Malik⁵, P. Reddy⁶;
¹Loughborough University, Loughborough, UNITED KINGDOM, ²Institute of Human Genetics, Newcastle Upon Tyne, UNITED KINGDOM, ³Human Biology Punjabi University, Patiala, INDIA, ⁴ISI, Calcutta, INDIA, ⁵SGSIPU, Delhi, INDIA, ⁶Oregon Health Sciences University, Portland, OR.

ALU polymorphisms provide a useful tool to population geneticists for understanding the population dynamics that have occurred over time. We report here a study of Six Alu insertion loci (TPA25, D1, APO, PV92, FXIIIB and ACE) from 20 endogamous caste and tribal populations of India and 5 regionally subdivided populations of Britain. Overall spectrum of variation in these populations is very interesting at different geographical and cultural levels. High level of insertion frequencies was observed in some highly inbred groups. Average levels of heterozygosities were found to be relatively high in these populations (range 41% to 49.8%). The genetic diversity coefficient GST among this group of populations was observed to be high. Phylogenetic trees and principal components analysis (PCA) computed from Alu frequencies provide support for socio-cultural and geographical assignment of these populations in Indian population structure. Comparisons are made with other world populations to understand genetic diversity and dynamics of Alu variation in British and Indian populations.

Population genetic analysis of CAG repeats of IT15 in populations of Volga-Ural region of Russia

I. A. Kutuyev¹, I. M. Khidijatova¹, R. I. Fatkhlislamova¹, R. V. Magzhanov², E. K. Khusnutdinova¹;
¹Institute of Biochemistry and Genetics of Ufa Science Center of Russian Academy of Sciences, Ufa, RUSSIAN FEDERATION, ²Bashkir State Medical University, Ufa, RUSSIAN FEDERATION.

The mutation causing Huntington disease (HD) is expansion of a trinucleotide repeat in the 5' end of IT15 gene on 4p16.3 beyond the normal range of 35 repeats. The investigation addresses genetic factors associated with normal variation of the CAG repeat polymorphism in HD gene. In the study blood samples of 602 individuals from 8 populations from Volga-Ural region were investigated: 4 ethnic groups of Bashkirs, Tatars, Russians, Chuvashes, Mordvinians, Maris, Udmurts, and Komis. The method involved PCR amplification using primers flanking only CAG polymorphic cite (without CCG repeats) with subsequent PAGE and dyeing with ethidium bromide. In all populations normal distribution of CAG repeats was found (p<0,001) showing unimodal nature peaking around repeat CAG17; the distribution skewness varied from 0.74 in Chuvashs to 1.98 in Bashkirs. 26 alleles containing from 9 to 36 repeats were revealed in total sample. A new sporadic case of HD was revealed in a person clinically silent and without positive family history of the disease with genotype CAG17/36. Fst, Fis, Fit, Gst, Dst were calculated. Four populations showed rather high rate of intermediate alleles (CAG29-35): Russians, Chuvashes, Mordvinians, and Udmurts (0.06, 0.01, 0.02, and 0.01 subsequently). We found extremely heterogeneity of investigated populations as in comparison with each other so with others world populations (Caucasians, Asians, and African Blacks). Total observed and expected heterozigosity for the locus were 0.751 and 0.769 respectively. Almost all populations were in HW equilibrium.

The impact of prevention strategies on the prevalence of neural tube defects in Hungary, 1987-1999

J. Metneki, K. Szalma, M. Szunyogh, C. Siffel;
National Center for Epidemiology, Budapest, HUNGARY.