[DNA profiling as a method of zygosity determination in multiple pregnancy]. / Profilowanie DNA jako metoda okreslania zygotycznosci w ciazy wieloraczej.

The aim of the report was to present a method of zygosity determination in multiple pregnancy. The study wascarried out on same-sex neonates born as a result of spontaneous quadruplet pregnancy. Zygosity was determinedby DNA profiling. The pregnancy was confirmed to be polyzygotic. DNA profiling may be used as a method ofzygosity determination in multiple pregnancy.

Frequency of streptococcal upper respiratory tract infections and HLA-Cw*06 allele in 70 patients with guttate psoriasis from northern Poland.

INTRODUCTION: The association of guttate psoriasis with a streptococcal throat infection and HLA-Cw*06 allele is well established in different populations. Nevertheless, only few studies on this form of disease have been performed in the Polish population. AIM: To analyze the frequencies of streptococcal-induced guttate psoriasis and HLA-Cw*06 allele in 70 patients with guttate psoriasis originating from northern Poland. MATERIAL AND METHODS: Seventy patients with guttate psoriasis and 24 healthy volunteers were enrolled into the study. Both groups were sex- and age-matched. The evidence of streptococcal infection was based on the positive throat swabs and/or elevated ASO titers. The modified method, including PCR-SSP and PCR-RFLP, was applied to HLA-Cw*06 genotyping. RESULTS: HLA-Cw*06 allele was confirmed in 49 (70%) out of 70 patients, which is significantly higher than in the control population (30%) (p = 0.001). Evidence for streptococcal infection was found in 34 (48.5%) subjects with psoriasis. Twenty-seven of them (79%) carried HLA-Cw*06 allele. In 36 individuals in whom no evidence of streptococcal infection was found, 14 (39%) did not carry HLA-Cw*06 allele. CONCLUSIONS: Our data confirm that HLA-Cw*06 is a major, but not imperative, genetic determinant for guttate psoriasis.

[Identification of a novel Y-SNP in the USP9Y gene and its impact on genotyping alleles of the M46 locus]. / Identyfikacja polimorfizmu typu Y-SNP w genie USP9Y i jego znaczenie w genotypowaniu alleli locus M46.

Y-chromosomal SNP markers are becoming increasingly more popular among forensic geneticists, but since they constitute variants specific to the ethnic origin, detailed population studies are required. Research into frequency of haplogroup N-M46 in the Belarusian population detected a mutated allele in 22 males, including one with a very distinct Y-STR haplotype. Sequencing of the M46 locus of this individual revealed the presence of a novel Y-SNP nearby the M46 locus, which was responsible for the erroneous assignment of the Y chromosome to the haplogroup N-M46. An impact of the identified polymorphism on discrimination of alleles of the M46 locus with various techniques was discussed, and solutions ensuring correctness of the genotyping results were proposed.

[Analysis of mutations in father-son pairs within selected Y-STR loci]. / Analiza czestosci mutacji w parach ojciec-syn w wybranych markerach Y-STR.

The objective of the study was to examine the mutation rates of Y-chromosomal STR from father-son pairs. The paternity in these cases was confirmed previously with the use of autosomal STR system performing standard analyses of genetic profiles of the mother, child and putative father (PI > = 100000). We examined 200 father-son sample pairs from Northern Poland using the Y-STR 18-plex. We identified eleven mutations. Five mutations resulted in the gain of a repeat in the sons' chromosome and six resulted in a loss of a repeat. All the samples resulted in single repeat mutations from one sample, which contained a two repeat loss at DYS385. The overall average mutation rate estimate was 0.0031.There was no significant difference in the mutation rate between Y-STR loci of the 200 tested father-son pairs and the YHRD base.

[Examples of application of X chromosomal markers in familial investigations and personal identification]. / Przyklady zastosowania markerów chromosomu X w badaniach rodzinnych i identyfikacyjnych.

Besides autosomal STR loci, markers of sex chromosomes, X and Y, are increasingly more commonly used in genetic analyses aiming at paternity testing or personal identification. The paper presents cases in which analysis of microsatellite loci of the X chromosome (X-STRs) was included in the routine examination and allowed for an unambiguous determination of the relationship between the tested individuals. The cases addressed paternity testing of female children, determination whether the examined women were paternal half-sisters, as well as personal identification of a deceased man. In none of the conducted expert opinions, the putative father's DNA sample was't available. Genotyping of X-STR markers was carried out with the use of commercial kits: Mentype Argus X-8 PCR Amplification Kit (Biotype) and Investigator Argus X-12 Kit (Qiagen).

[Personal identification of an unknown individual based on determination of his DNA profile from exhumed remains]. / Ustalenie tozsamosci nieznanej osoby w oparciu o okreslenie profilu DNA z ekshumowanych szczatków ludzkich.

The aim of the present investigation was personal identification of an unknown man whose remains were exhumed four years after burial. The femur of the deceased was secured for the genetic analysis. The comparative material included buccal swabs collected from the putative relatives of the deceased, i.e. the wife, son and brother. Genomic DNA was extracted from the bone using two methods: traditional isolation with phenol/chloroform and as a alternative technique, a simple and rapid method described by T. Kalmár et al. The results were then compared. The specimens underwent DNA amplification using the AmpFISTRSEfiler PCR Amplification Kit. The authors obtained a full STR profile of the unknown man from each isolate, yet the DNA extraction method proposed by T. Kalmár et al. allowed for simpler and faster isolation of genetic material. The statistical analysis of the obtained results confirmed the paternity of the deceased and established his son as his rightful child (P = 99.999999%), also confirming the consanguinity between the investigated individual and his putative brother (P = 99.9999%).

Frequency of DRB1-DQB1 two-locus haplotypes in tuberculosis: preliminary report.

Analysis of correlation between tuberculosis (TB) and human leukocyte antigen (HLA) in populations from Asia and Latin America has shown conflicting results. The aim of this study was to evaluate the frequency of HLA-DRB1-DQB1 two-locus haplotypes of 61 TB patients and 125 healthy volunteers in the same ethnic group in Poland. DRB1 and DQB1 alleles were determined by PCR-SSP "low-resolution" and "high-resolution" methods. Our study showed that DRB1*1601 and DQB1*0502 alleles were more frequent, whereas DQB1*0201 was rarer in TB than in controls. DRB1*16-DQB1*05, DRB1*04-DQB1*03 and DRB1*1601-DQB1*0502 haplotype were more common, and DRB1*11-DQB1*03 less frequent in TB in comparison to controls. Positive linkage disequilibrium (LD) for DRB1*01-DQB1*05, DRB1*03-DQB1*02, DRB1*11-DQB1*03, DRB1*13-DQB1*06 and DRB1*15-DQB1*06 was found in controls. A trend towards the positive LD for DRB1*01-DQB1*05, DRB1*03-DQB1*02, DRB1*11-DQB1*03, DRB1*15-DQB1*06 and DRB1*16-DQB1*05 was shown in TB. The trend towards the positive LD for DRB1*16-DQB1*05 haplotype in TB patients was not observed in the control group. It seems likely that the presence of DRB1*1601, DQB1*0502 alleles and DRB1*1601-DQB1*0502, DRB1*04-DQB1*03, DRB1*14-DQB1*05 haplotypes may be related to a higher risk of developing TB, whereas low frequency of DQB1*0201 and DRB1*11-DQB1*03 haplotype may be linked to the resistance to TB.

[Polymorphism of X-STR loci DXS7108 and DXS1196 in the Northern Polish population]. / Polimorfizm chromosomu X--STR loci DXS7108 i DXS1196 w populacji Polski pólnocnej.

This paper describes results of a population study of two X-linked STR microsatellite markers: DXS7108 and DXS1196. 298 samples of DNA of unrelated persons (male and female) from the Northern part of Poland were analyzed. DNA was isolated using a non-enzymatic method. After amplification PCR products were separated by means of capillary electrophoresis using the ABI PRISM 310 Genetic Analyzer. The most common alleles of each locus were sequenced and used as a control ladder to type unknown samples. Testing for Hardy-Weinberg equilibrium (HWE) showed no significant deviation for these two loci. Statistical parameters (PD, HET, MEC) showed that examined systems are useful in forensic medicine.

Variation of X-chromosomal microsatellites in Belarus within the context of their genetic diversity in Europe.

More and more X-STR data are becoming available for worldwide human populations for forensic and anthropological investigations, but the European datasets analysed so far represent mainly the central, northern, western and southern part of the continent with populations of Eastern Europe being practically uninvestigated. In the present study, we assessed genetic variation and linkage disequilibrium of 19 X-chromosomal STR markers (DXS7132, DXS7133, DXS7423, DXS7424, DXS8377, DXS8378, DXS9895, DXS10074, DXS10075, DXS10079, DXS10101, DXS10103, DXS10134, DXS10135, DXS10146, DXS10147, DXS10148, GATA172D05, HPRTB) in four regional populations of an Eastern European state of Belarus, including 12 loci incorporated in the Argus X-12 kit. Our results revealed cumulative power of discrimination of the tested X-STR loci to amount to 0.999999999999996 and 0.999999997 in females and males, respectively. Analysis of molecular variance demonstrated regional stratification within the country, excluding the use of a common X-STR database for Belarus in forensic casework. However, development of a separate X-STR database for the northwestern part of the country or exclusion of four loci displaying regional differences from the dataset were shown to eliminate the observed geographic substructure among Belarusians. Comparison of the Belarusian genotypes with X-STR data from other European populations disclosed a geography-driven northeast-southwest gradient extending from Belarus and Finland to Iberia and Italy. This study is the first extensive report on variation of X-STR markers in populations from Eastern Europe and the first comprehensive analysis of diversity of X-chromosomal microsatellites in Europe.

Comparative analysis of HLA class I antigens in pulmonary sarcoidosis and tuberculosis in the same ethnic group.

OBJECTIVE: To compare occurrences of respective HLA class I antigens in patients with sarcoidosis (SA), patients with tuberculosis (TB), and healthy controls in the same ethnic group in Poland. PATIENTS AND METHODS: HLA-A, -B, and -C antigens were determined from 1994 to 1997 by using the National Institutes of Health method in 100 patients with SA, 100 patients with TB, and 100 healthy controls. Frequencies of specific antigens were compared among the 3 groups. RESULTS: Our study showed that among SA patients, the HLA-B51(5) and HLA-B8 antigens tended to be more common and the HLA-B13, -B35, and -Cw4 antigens tended to be less common than in the controls. However, after Bonferroni correction, only the HLA-B35 antigen was found to be significantly different in SA patients and controls. In TB patients, the expression of HLA-B62(15) and HLA-Cw5 antigens tended to be more common compared with controls and HLA-A2 less common compared with controls, but only the differences in B62(15) and Cw5 were significant after Bonferroni correction. HLA-B51(5) and HLA-B8 antigens were statistically more frequent and B13, B62(15), and Cw4 less frequent in SA than in TB patients and remained significant after Bonferroni correction. The occurrence of other antigens studied in both populations was comparable. CONCLUSION: We identified associations of HLA class I antigens in patients with SA or TB, with an expression pattern specific and different for each group.

Analysis of polymorphism of three human Y-chromosome STR loci: DYS390, DYS392 and DYS393 in the population of northern Poland.

The study aimed at development of a multiplex PCR system for amplification of three Y-chromosome STR loci: DYS390, DYS392 and DYS393, and its application in haplotype polymorphism analysis in the population of northern Poland. Due to interactions between originally published primers, a new DYS392 primer pair was proposed. In a population of 158 unrelated males, 28 different haplotypes could be observed, 12 of which were seen only once. The haplotype diversity is 0.805. Distribution of haplotypes of the studied loci is specific to the population of northern Poland and distinguishes it from compared West-European populations. To our knowledge, this is the first report on a Y-STR multiplex system that can be analysed on native polyacrylamide gels.

Correlation of HLA-Cw*06 allele frequency with some clinical features of psoriasis vulgaris in the population of northern Poland.

Psoriasis is a common skin disease with a genetic background and significant human leukocyte antigen (HLA) associations. HLA-Cw6 is the most frequently described association, particularly with psoriasis of the early onset type. Few studies of its correlation with various psoriasis clinical phenotypes and severity of the disease have been published so far; none in the Polish population. In this study 78 patients with psoriasis vulgaris were evaluated clinically and subdivided according to the age of onset and the type of psoriasis. A system of disease severity evaluation was applied to each patient. All patients and the control group (70 unrelated persons) were typed for HLA-Cw*06. The results show that Cw*06 allele frequency was higher in psoriatics than in the control group. Our investigation confirms that the Cw*06 allele is positively associated with psoriasis vulgaris of the early onset type, with a positive family history and its more severe form.

[Application of polymorphism of HLA-DR locus in personal identification]. / Mozliwosci wykorzystania polimorfizmu locus HLA-DR w identyfikacji osobniczej.

The aim of this paper was to present the results of investigation of the high polymorphic HLA-DR locus by the PCR-SSP method. Genomic DNA was isolated from whole blood, tissue samples (muscles and brains). HLA-DR alleles were typed using the DYNAL DR "low resolution" SSP kit. PCR products were separated on agarose gel with ethidium bromide and estimated in ultraviolet light. Phenotypic and genotypical rates of blood and tissues have been analysed. After electrophoresis in 228 DNA samples 39 phenotypes and 10 alleles were identified. The most frequent phenotypes of the HLA-DR locus were 5/6, 6/6 and 3/5. The high frequency of HLA-DR 6, DR 5 and DR 7 alleles have been found. No deviations from the Hardy-Weinberg equilibrium were observed. The HLA-DR locus has a relatively high information content (PIC), power of discrimination (PD) and mean exclusion chance (MEC). The molecular PCR-SSP method allows for very rapid and highly discriminatory HLA analysis. We found the PCR-SSP technique very useful in routine HLA-DR identification.

[Identification of a very short YCAII allele in the northern Polish population]. / Identyfikacja bardzo krótkiego allela YCAII w populacji pólnocnej Polski.

The YCAII system is one of the Y-chromosomal STR markers with dinucleotide repetitive units and due to the duplication on the chromosome, its amplification provides two polymorphic products. Due to the high polymorphism, YCAII was included in the Y-STR Haplotype Reference Database (http://www.ystr.org) and significantly increased discrimination power of the collected haplotypes. During studies on the Y-STR polymorphism in the Polish population, a very short YCAII allele with relatively low stutter was identified. Sequencing revealed the presence of 11 dinucleotide repetitive units characteristic of this system. Among 3998 extended haplotypes collected in the database, this allele was found only in the population of south-western Germany. Similarities between the haplotypes possessing the YCAII*11 allele suggest their common origin.

[Polymorphism of the DXS1062 locus in a Polish population]. / Polimorfizm locus DXS1062 w populacji polskiej.

This paper presents the results of a population study of a dinucleotide STR marker DXS1062. Blood samples were obtained from unrelated adult individuals (males and females) living in the northern part of Poland. In the analyzed population, 21 different phenotypes and 9 alleles of the DXS1062 locus were found. The alleles were sequenced and used for the construction of an allelic ladder. The nomenclature in accordance with ISFG guidelines was proposed. The most frequent alleles were 20 and 21. Statistical parameters (PR, PM, PD, PIC) showed that the examined system is useful in forensic medicine.

[Entomological methods of determining time of death]. / Entomologiczne metody okreslania czasu smierci.

The paper contains descriptions of the entomological methods which can be used in determining time of death. It based on the knowledge of groups of arthropods (waves of succession), characteristic for stages of decomposition of the corpse as well as the biology and longevity of development of necrophagous insects.

[Effect of bovine serum albumin and restriction enzyme on quality of DNA profile of tissues extracted from paraffin blocks and histological slides]. / Wplyw stosowania albuminy wolowej i enzymu restrykcyjnego na jakosc profili DNA tkanek wyizolowanych z bloczków parafinowych i z preparatów histologicznych.

This paper reports the possibility of DNA profiling obtained from fixed and paraffin-embedded tissues or histological slides. The influence of using bovine serum albumin and restriction enzyme Hinfl on quality of DNA was analysed. The usability of those chemicals was estimated by examination of amplification results locus VWA, and multiplex PCR with the use commercial kit AmpFlSTR Identifiler (including 15 loci + amelogenine locus) and AmpFlSTR SEfiler kit (including 11 loci + amelogenine locus) Applied Biosystems. PCR products of VWA locus were separated by electrophoresis on polyacrylamide gels and visualized by silver staining. The products of loci of AmpFlSTR Identifiler and AmpFlSTR SEfiler were analyzed by capillary electrophoresis on an ABI Prism 310 sequencer. Results achieved with and without using bovine serum albumin and restriction enzyme Hinfl were compared and positive influence of those substances on DNA quality was demonstrated. Obtained results of purification DNA with the use of restriction enzyme were verified in paternity testing case.

[The use of entomologic methods in determining the time of death--case studies]. / Wykorzystanie metod entomologicznych w próbach okreslenia daty zgonu--opis przypadków.

This paper describes a number of cases in which the determination of the time of death was based on entomological methods. These cases took place in the Pomorskie Province, and the autopsies were carried out in the Department of Forensic Medicine, Medical University of Gdansk.

[Determination of polymorphic DNA sequences STR-PCR in decomposed human tissues]. / Okreslanie polimorficznych sekwencji DNA typu STR-PCR w rozlozonych tkankach ludzkich.

This paper reports the possibility of DNA profiling obtained from soft tissues influenced by the decomposition process: heart, kidney, liver collected during autopsy. As a control DNA, profile from blood was determined. DNA was extracted by the phenol-chloroform method. Amplification was performed with the use of the GenePrintSTR Multiplex (CSF1PO, TPOX, TH01) system, Promega and AmpF[symbol: see text]STR Identifier, Applied Biosystems. PCR products of the CTT system were separated by electrophoresis on denaturing polyacrylamide gels and visualized by silver staining. The 16 loci of AmpF[symbol: see text]STR Identifier products were analyzed by capillary electrophoresis on an ABI PRISM 310 sequencer. The results from both methods were compared. Electrophoresis of the CTT products showed clear results for DNA extracted from blood and decomposed tissues, particularly from the heart and kidney. The capillary electrophoresis method gave a positive signal for all 16 loci of AmpF[symbol: see text]STR Identifier for DNA extracted from heart, kidney and blood. Worse results similar to the manual method were obtained for DNA extracted from the liver. The soft tissues, also decomposed by putrefaction can be a useful source of genomic DNA in personal identification and paternity testing.

Northern Slavs from Serbia do not show a founder effect at autosomal and Y-chromosomal STRs and retain their paternal genetic heritage.

Studies on Y-chromosomal markers revealed significant genetic differentiation between Southern and Northern (Western and Eastern) Slavic populations. The northern Serbian region of Vojvodina is inhabited by Southern Slavic Serbian majority and, inter alia, Western Slavic (Slovak) and Eastern Slavic (Ruthenian) minorities. In the study, 15 autosomal STR markers were analysed in unrelated Slovaks, Ruthenians and Serbs from northern Serbia and western Slovakia. Additionally, Slovak males from Serbia were genotyped for 17 Y-chromosomal STR loci. The results were compared to data available for other Slavic populations. Genetic distances for autosomal markers revealed homogeneity between Serbs from northern Serbia and Slovaks from western Slovakia and distinctiveness of Serbian Slovaks and Ruthenians. Y-STR variation showed a clear genetic departure of the Slovaks and Ruthenians inhabiting Vojvodina from their Serbian neighbours and genetic similarity to the Northern Slavic populations of Slovakia and Ukraine. Admixture estimates revealed negligible Serbian paternal ancestry in both Northern Slavic minorities of Vojvodina, providing evidence for their genetic isolation from the Serbian majority population. No reduction of genetic diversity at autosomal and Y-chromosomal markers was found, excluding genetic drift as a reason for differences observed at autosomal STRs. Analysis of molecular variance detected significant population stratification of autosomal and Y-chromosomal microsatellites in the three Slavic populations of northern Serbia, indicating necessity for separate databases used for estimations of frequencies of autosomal and Y-chromosomal STR profiles in forensic casework. Our results demonstrate that regarding Y-STR haplotypes, Serbian Slovaks and Ruthenians fit in the Eastern European metapopulation defined in the Y chromosome haplotype reference database.