In search for the grave of 100 Poles executed on March 20, 1942 in Zgierz, Poland - research by SIGO (Network for Genetic Identification of Victims).

It can be reasonably assumed that remains exhumed in 2012 and 2013 during archaeological explorations conducted in the Lucmierz Forest, an important area on the map of the German Nazi terror in the region of Lodz (Poland), are in fact the remains of a hundred Poles murdered by the Nazis in Zgierz on March 20, 1942. By virtue of a decision of the Polish Institute of National Remembrance's Commission for the Prosecution of Crimes Against the Polish Nation, the verification of this research hypothesis was entrusted to SIGO (Network for Genetic Identification of Victims) Consortium appointed by virtue of an agreement of December 11, 2015. The Consortium is an extension of the PBGOT (Polish Genetic Database of Totalitarianisms Victims). So far, the researchers have retrieved 14 DNA profiles from among the examined remains, including 12 male and 2 female profiles. Furthermore, 12 DNA profiles of the victims' family members have been collected. Due to the fact that next-of-kin relatives of the victims of the Zgierz massacre are of advanced age, it is of key importance to collect genetic material as soon as possible from the other surviving family members, identified on the basis of a list of victims that has been nearly completely compiled by the Polish Institute of National Remembrance (IPN) and is presented in this paper.

Polymorphism of 12 STR loci included in the Investigator HD-plex kit in Polish population of Lodz region.

In this study Polish population data as well as efficiency parameters of 12 STR loci included in the Investigator HDplex set were presented. This set contains 9 systems not available in any other commercial multiplexes, ie.: D2S1360, D3S1744, D4S2366, D5S2500, D6S474, D7S1517, D8S1132, D10S2325 and D21S2055. The evaluation was preformed based on DNA samples derived from 303 unrelated individuals living in Lodz region, central part of Poland. The obtained distribution of the genotypes is consistent with the assumptions of the Hardy and Weinberg equilibrium (HWE). It reflects properly genetic structure of the studied population compared with other populations of Europe and the world. It indicates the linkage equilibrium within the pairs of investigated loci, as well as with regard to other syntenic loci. The total value of the power of exclusion (PE) and the random match probability (MP) were respectively 0.99999988 and 5.2 × 10-18. Therefore the polymorphism of examined genetic markers within the Investigator HD-plex multiplex allows for a significant increase of the evidence value. Thus it constitutes an excellent tool for resolving difficult cases in the field of forensic genetics.

Population database on: D1S1656, D2S441, D2S1338, D3S1358, D8S1179, D10S1248, D22S1045, D12S391, D16S539, D18S51, D19S433, D21S11, FGA, TH01, vWA loci included in NGM system based on one thousand unrelated individuals from Lodz region of Central Poland

A population data obtained on the basis of sample of 1000 unrelated individuals of Polish ancestry living in Lodz region of Central Poland with use of fluorescent multiplex-PCR and capillary electrophoresis were presented. Evaluation included 15 polymorphic loci DNA - STR from NGM multiplex-PCR set, ie. D1S1656, D2S441, D2S1338, D3S1358, D8S1179, D10S1248, D12S391, D16S539, D18S51, D19S433, D21S11, D22S1045, FGA, TH01, vWA. The allele frequency distribution and crucial statistical parameters for the investigated markers and the whole set were calculated. The compliance of the studied population with Hardy-Weinberg equilibrium, independence of inheritance and high parameters of the usefulness in forensic genetics have been demonstrated. The interpopulation comparison performed by the "neighbor-joining" method as well as multidimensional scaling depicted the genetic distances dividing the examined Polish population from other populations of Poland, Europe and the world.

Recommendations of the Polish-speaking Working Group of the International Society for Forensic Genetics (ISFG-PL) regarding the disclosure of biological traces and the handling of evidence for identification tests. / Zalecenia Polskojezycznej Grupy Miedzynarodowego Towarzystwa Genetyki Sadowej (ISFG-PL) dotyczace ujawniania i identyfikacji sladów biologicznych przeznaczonych do badan genetycznych.

The purpose of this paper is to formulate recommendations for the disclosure of biological traces in the laboratory and the handling of forensic evidence submitted for identification tests, recommended by the Polish Speaking Working Group of the International Society for Forensic Genetics. The paper organizes the knowledge of the most relevant stages of preliminary analysis of biological traces based on both literature sources and those resulting from years of research practice. Recommendations formulated in the course of multi-stage expert consultations contained in this study should be used in the development of laboratory procedures applied during the execution.

Forensic evaluation of the AmpFlSTR® NGM™ loci in Lodz region of Poland population sample.

The paper is focused on population data for 15 polymorphic STR loci included in the NGM(TM) amplification kit, obtained from a sample of 800 individuals from the Lodz region of Poland. Main statistical parameters of forensic interest were calculated and Hardy-Weinberg equilibrium was verified for each locus. Departure from HWE was not significant after applying Bonferroni corrected significance level for multiple testing (p = 0.0033). Comparative analysis between chosen populations was performed and some significant differences were found among investigated populations. Obtained values of parameters for NGM™ multiplex amplification kit point to wide range of possible applications of investigated STR markers to forensic genetics.

Genetic investigation of biological materials from patients after stem cell transplantation based on autosomal as well as Y-chromosomal markers.

The authors presented the results of DNA polymorphism investigation of blood, buccal swabs and hair follicles originating from patients after allogeneic hematopoietic stem cell transplantation. The real-time and multiplex assays based on polymerase chain reaction within the range of autosomal as well as Y-chromosomal markers were applied to assess the possible dangers arising from investigation of these materials in forensic genetics. The results revealed that not only post-transplant blood and buccal swab, but also recipient hair, up to now regarded as devoid of any donor's cells, do not constitute entirely safe material for forensic purposes. Their analysis can lead to the false identification of gender or male haplotype. The investigation of sex-determining region Y and Y-chromosome short tandem repeats performed in female recipients with male donors resulted in the designation of donor's DNA in hair cells as well as in blood and buccal swabs. Therefore, biological stains gathered from crime scenes should not be analysed exclusively based on the investigation of male-specific markers.

[Genetic analysis of human remains exhumed during archaeological excavations on former military training ground Brus in Lodz]. / Analiza genetyczna szczatków ludzkich ekshumowanych podczas badan archeologicznych na terenie bylego poligonu na Brusie w Lodzi.

The aim of this study was the genetic identification of Nazi repression victims. Human remains were found in 2011 in the area of former military training ground BRUS in Lodz. Genetic tests were performed upon the request of the Departmental Commission for the Prosecution of Crimes against the Polish Nation of the Institute of National Remembrance in Lodz. The research material was provided by the Institute of Archaeology (University of Lodz). It consisted of bones and teeth which were exhumed from mass Grave No 7. As a reference material we used a buccal swab collected from the putative son of one of the victims. Genomic DNA was extracted from the skeletal samples using the PrepFiler BTA Forensic DNA Extraction Kit. DNA was amplified using the AmpFlSTR Identifiler Plus PCR Amplification Kit and analyzed using an AB 3500 genetic analyzer. The obtained results showed 12 male genetic profiles. The analysis excluded paternity of 10 investigated victims. The genetic data of the remaining samples did not allow for paternity settlement.

[Y-STR Poland--a database for evaluation of evidence value in forensic genetics]. / Y-STR Polska--baza danych do oceny wartosci dowodowej w genetyce sadowej.

The "Y-STR Poland" is a multicenter project, the aim of which is the construction of a widely available database of Y chromosome haplotypes determined in the Polish population in a range of sixteen loci in AmpFISTR Y-filer system. The database will be regularly updated and it will be used in assessment of evidence value in forensic genetics. The starting base "Y-STR Poland" contains 1600 Y-STR haplotypes and encompasses data collected in Lodz (two independent centers), Warsaw and Szczecin regions. The present report contains as an attachment the data in an Excel-type file, which serves as a tool in frequency determination of a given Y haplotype in the Polish population. The file will be updated on a regular basis along with updating the database, and will be freely available from www.genetyka-sadowa.pl.

Examination of LT-DNA traces - literature overview and general recommendations of the Polish Speaking Working Group of the International Society for Forensic Genetics (ISFG-PL).

The available literature on traces characterised by a suboptimal amount of DNA, as well as expert research practice, show the complex nature of LT-DNA traces: from their detection and collection, through genetic analysis, up to the interpretation of final results. The aims of this paper are to systematise the current state of knowledge on handling LT-DNA traces and develop examination guidelines, as recommended by the Polish Speaking Working Group of the International Society for Forensic Genetics (ISFG-PL). The proposed guidelines should be followed by all Polish laboratories conducting forensic genetic analyses for the purpose of judicial proceedings.

Recommendations of the Polish Speaking Working Group of the International Society for Forensic Genetics on forensic Y chromosome typing.

Y chromosome typing has been performed in forensic genetic practice for more than 20 years. The latest recommendations of the DNA Commission of the International Society of Forensic Genetics (ISFG) concerning the application of Y-chromosomal markers in forensic genetics were published in 2006. The aim of this report is to recapitulate, systematise and supplement existing recommendations on the forensic analysis of polymorphism of the Y chromosome with standards already implemented in practice, new capabilities linked to the development of research techniques as well as current solutions used in statistical analysis. The recommendations have been adapted specifically to aspects related to the preparation of expert opinions in the field of forensic genetics in Poland. The Polish Speaking Working Group of the ISFG believes that the presented guidelines should become a standard implemented by all Polish laboratories performing Y chromosome typing for forensic purposes.

[The usefulness of SNP markers for analyses of highly degraded biological materials]. / Przydatnosc markerów SNP do analiz materialu biologicznego o wysokim stopniu degradacji.

The most common cause of problems associated with analyzing DNA extracted from forensic samples is their high level of degradation. Such difficulties are caused by the fact that STR markers have too large amplicon sizes to be amplified in degraded DNA samples. Thus, it is necessary to employ more efficient markers for analyzing evidential samples. SNPs are ideal tools for such purposes, for the SNP genotyping method does not require large amplicon size, and thus increases the possibility of amplifying degraded DNA samples. Although single SNP is not polymorphic enough, we can obtain sufficient results by examining several SNPs. The aim of this study was to examine the usefulness of the SNP-pentaplex (rs2294067, rs2282160, rs2070764, rs2277216, rs1063739) for forensic applications by analysing several forensic cases, which were impossible to solve in a range of STR markers because of highly degraded DNA. DNA fragments were amplified in one multiplex PCR reaction, which contained 5 primer pairs. SNPs were subsequently identified in a minisequencing reaction and gel electrophoresis in an ABI Prism 377 Sequencer. The research confirmed the usefulness of SNP-pentaplex for forensic applications. Despite employing mainly degraded and low copy number DNA, full genetic profiles were obtained in almost every sample. Although the discrimination power of SNP-pentaplex is not sufficient for obtaining adequate evidential value, it seems to be an ideal screening method for forensic applications.

[Application of the QIAamp DNA Investigator Kit and Prepfiler Forensic DNA Extraction Kit in genomic DNA extraction from skeletal remains]. / Wykorzystanie zestawów QIAamp DNA Investigator Kit oraz PrepFiler Forensic DNA Extraction Kit w procesie izolacji genomowego DNA z materialu kostnego.

The report presents an application of the QIAamp DNA Investigator Kit and PrepFiler Forensic DNA Extraction Kit in genomic DNA extraction from post-mortem highly degraded skeletal remains. The analysis included 25 bone samples collected on autopsy. DNA extraction was performed in accordance with the QIAamp DNA Investigator Kit and PrepFiler Forensic DNA Extraction Kit manufacturer's isolation protocols. Amplification was performed on a Biometra termocycler using the AmpFISTR Identifiler PCR Amplification Kit according to the manufacturer's protocol. Typing of PCR products was carried out on an ABI Prism 377 DNA sequencer. The recommended parameters for GeneScan analysis and Genotyper software were followed. The authors demonstrated that the QIAamp DNA Investigator Kit was more effective, convenient and statistically significantly better method which may be employed in DNA extraction from bone specimens.

Sequence analysis of a new short tandem repeat locus D17S2266E located in the human growth hormone gene cluster HumGH@.

D17S2266E is a new, variable genetic marker exhibiting polymorphism of the number of repeats of four- and two-nucleotide motifs. This study, carried out on a group of 250 unrelated persons from various regions of Poland, revealed the presence of 24 different alleles ranging in size from 232 to 290 base pairs. Analysis of the sequenced fragments demonstrated that the alleles consisted of two flanking regions and two variable blocks that were separated by a consensus sequence. There were (AAAG)(5)(AG)(1)(AAAG)(3-4) repeats in the first block, and [(AAAG)(2)(AG)(1)](0-1)[(AAAG)(6)(AG)(1)](0-1)(AAAG)(n) repeats in the second block. On the basis of the allele frequencies in the population, we were able to do biostatistical calculations, which gave the following results: expected heterozygosity 0.8947 +/- 0.0137, power of discrimination 0.9793, polymorphism information content 0.8837, probability of exclusion (PE) 0.7859, PE for motherless cases 0.6473, and an average paternity index of 4.7470. These biostatistical parameters show that the marker D17S2266E can find a wide range of applications in forensic testing.

[Association of the tyrosine hydroxylase gene polymorphism with schizophrenia in the population of central Poland]. / Asocjacja miedzy polimorfizmem genu hydroksylazy tyrozyny a wystepowaniem schizofrenii w populacji Polski centralnej.

AIM: The aim of the study was a comparative analysis of the polymorphism in TH01 locus-- tetranucleotide microsatellite region located in the first intron of the tyrosine hydroxylase gene (TH) between a group of Polish patients suffering for schizophrenia and their regionally matched healthy subjects. METHOD: One hundred patients affected by paranoid schizophrenia and healthy individuals with negative family history of psychiatric disorders as control, were investigated. Genomic DNA was isolated from peripheral blood, amplification of TH01 locus was carried out using the following sequences of primers: 5'-GTG GGC TGAAAAGCT CCCGAT TAT-3', 5'-ATT CAA AGG GTA TCT GGG CTC TGG-3', PCR products were detected on the ABI Prism 377 sequencer. Distributions of alleles, genotypes and homo-heterozygosity of the patients were compared with those of the matching controls using the RXC program created by G. Carmody. Relative Risk (RR) of the disease was calculated according to the formula given by Dyer & Werrens. RESULTS: . The conducted analysis showed the existence of statistically significant differences in the distribution of alleles, as well as genotypes between the schizophrenics and the control population. We revealed that 7 allele is present statistically significantly more often in the group of patients and its presence increases the risk of schizophrenia almost twofold (RR = 1.89). Whereas the presence of 9.3 allele reduces the risk of the disease (RR = 0.72), in the homozygote form 9.3-9.3 even over three times (RR = 0 .32). CONCLUSION: The revealed differences in the susceptibility to schizophrenia depending on polymorphic allele variants in repetitive TCAT sequence in TH01 locus may be associated with the function of a regulatory element in the process of TH gene transcription. Our findings need further investigation in a larger sample of patients.

[Antioxidative enzymes--structure, properties, functions]. / Enzymy antyoksydacyjne--budowa, wlasciwosci, funkcje.

In physiological condition mammalian cells produce a small amount of reactive oxygen species (ROS) which influence biological processes. At high concentration ROS are the mediator of damage to lipids, protein and nucleic acids. Exposure to free radicals has led organisms to develop series of defense mechanism that can protect against oxidative stress. The defense mechanism is represented by non-enzymatic antioxidants and enzymatic antioxidants that are mainly represented by: superoxide dismutases (SOD), catalase (CAT) and, glutathione peroxidases (GPx), glutathione reductase (GR). This article presents short review about structure, properties and functions of the above enzymes in living cells.

[The central Poland population database of 500 SNP alleles]. / Baza 500 alleli SNP w populacji centralnej Polski.

SNP analysis is one of the most contemporary methods for personal identification in forensic genetics. It is increasingly more frequently used in forensic practice, especially for analyses of highly degraded DNA samples from crime scenes and thus it requires suitable population data. The aim of this study was to develop a central Poland population database consisting of 500 alleles in a range of 5 SNP biallelic loci (rs2294067, rs2282160, rs2070764, rs2277216, rs1063739). DNA fragments were amplified in one multiplex PCR reaction and SNPs were identified in a minisequencing reaction. The combined PD of the pentaplex was 0.9907147657. This makes the pentaplex a good screening method for forensic applications.

Standardization of research in the field of forensic genetics - the current state in the world and introduction to the guidelines in Poland with reference to the work of the Expert Team for Standards and Assessment in Forensic Genetics of the Polish Speaking Working Group of the International Society for Forensic Genetics (ISFG-PL).

The beginnings of forensic genetics, one of the most rapidly growing fields of research, can be traced to Great Britain in 1985. It appeared in Poland in 1989. It uses the most advanced technologies, including the investigation of the variability of the human genome through mass parallel sequencing, which help, among other things, to analyze features of human appearance and origin. These technologies coexist with well standardized techniques of multiplex short tandem repeat analysis based on capillary electrophoresis, which allows to obtain a unique individual profile at a minimal cost. Legislation, research standards and guidelines developed by opinion-forming institutions and expert teams play a key role in the field of genetic forensic examinations. This study presents the current normative state of this area. It precedes the presentation of guidelines concerning the main aspects of research in the field of forensic genetics in Poland, prepared by a team of experts gathered within the Polish Speaking Working Group of the International Forensic Genetics Society and the Forensic Genetics Committee of the Polish Society of Forensic Medicine and Criminology.

Recommendations of the Polish Speaking Working Group of the International Society for Forensic Genetics for forensic mitochondrial DNA testing.

Although mitochondrial DNA (mtDNA) testing has been used in forensic genetics only since the mid-1990s, forensic DNA laboratories have been recently increasing the range of mtDNA sequencing, employing new analytical approaches and methods of data analysis. Therefore, it seems fitting to gather and systematize existing recommendations in the field of mtDNA analysis for forensic purposes, and formulate a set of interpretative guidelines which are especially relevant in view of recent developments in the forensic casework. The starting point is the recommendations of the International Society for Forensic Genetics (ISFG) which, in the opinion of the Polish Speaking Working Group of the ISFG (ISFG- PL), should be followed by all Polish laboratories conducting forensic testing.

[SNP pentaplex--the allele frequency database of central Poland population]. / Pentapleks SNP - rozklad czestosci alleli w populacji centralnej Polski.

SNP analysis is among the most contemporary method for personal identification in forensic genetics, especially in analyses of untypical cases and in studies of degraded DNA samples from crime scenes. This paper shows the results of Central Poland population studies carried out using minisequencing and SNP-pentaplex, which contains 5 biallelic loci rs2294067, rs2282160, rs2070764, rs2277216 and rs1063739. DNA fragments were amplified in one multiplex PCR reaction and SNPs were identified by the minisequencing method. The combined PD was 0.9895253125, which makes the pentaplex useful for forensic applications.