Evaluation of the role of rs2227983 polymorphism of EGFR gene in the development of allergic asthma

The objective of the study: to study rs2227983 polymorphism of EGFR gene in patients with allergic asthma and healthy individuals.Subjects and Methods. 179 patients suffering from allergic asthma were included in the study. The diagnosis and degree of severity were established in accordance with the GINA recommendations. The Control Group included apparently healthy individuals (n = 217). Patients with allergic asthma underwent standard laboratory and instrumental examinations and DNA typing.Results. A statistically significant predominance of AG genotype frequency in the group of patients with allergic asthma, including women, versus the group of healthy individuals, was established. AG rs2227983 genotype of EGFR gene was found to be significantly more common in patients with mild and moderate allergic asthma including women, than in healthy individuals, including women.Conclusion. The association of rs2227983 polymorphism of EGFR gene with allergic asthma has been established. A homozygous GG genotype may play a protective role against the disease.

Assessment of the level of allelic polymorphism of SSR markers and genetic distances of some grape varieties of the South of Russia of different ecological-geographical groups

Представлены результаты оценки генетического разнообразия 24 местных сортов юга России, поддерживаемых на ампелографической коллекции ФГБУН «ВННИИВиВ «Магарач». ДНК-типирование сортов и оценка аллельного разнообразия выполнено с использованием 9 ядерных (nSSR) и 3 хлоропластных (cpSSR) микросателлитных локусов. Уровень полиморфизма nSSR локусов составил 100 %. Всего было идентифицировано 73 аллеля, в среднем 9.1 аллеля /локус. Минимальное количество аллелей идентифицировано в локусах ssrVrZAG64 и ssrZag83. Наибольшее количество аллелей выявлено в локусе ssrVvUCH29 (13 аллелей), диапазон размера которых составил 203 п.н. - 309 п.н. В результате анализа полиморфизма сpSSR-локусов идентифицировано 4 хлоротипа: А, В, С, D. Наиболее распростанен в группе изученных сортов хлоротип D (58 %). В статье обсуждается происхождение сортов на основе анализа их гаплотипов. По результатами анализа аллельного полиморфизма nSSR-локусов рассчитана матрица генетических дистанций, значения которой находились в диапазоне 0,33-0,94, построена дендрограмма, отражающая взаимоотношения между образцами. По степени генетического сходства выделились 3 основных кластера, в которых наблюдалась дифференциация или тенденция к дифференциации по эколого-географическим группам. The assessment results of genetic diversity of 24 local varieties of the South of Russia, maintained in the ampelographic collection of the FSBSI Institute Magarach are presented. DNA typing of cultivars and assessment of allelic diversity was performed using 9 nuclear (nSSR) and 3 chloroplast (cpSSR) microsatellite loci. The level of polymorphism of nSSR loci was 100%. A total of 73 alleles were identified with an average of 9.1 alleles per locus. The minimal number of alleles was observed in the ssrVrZAG64 and ssrZag83 loci. The biggest number of alleles was found in the ssrVvUCH29 locus (13 alleles), the size range of which was 203 bp-309 bp. As a result of polymorphism analysis of cpSSR loci, 4 chlorotypes were identified: A, B, C, D. Chlorotype D is the most widespread in the group of the studied cultivars (58%). The article discusses the origin of varieties based on the analysis of their haplotypes. Based on the results of the analysis of allelic polymorphism of nSSR loci, a matrix of genetic distances was calculated, the values of which were in the range of 0.33-0.94, and a dendrogram, reflecting the relationship between the samples, was constructed. According to the degree of genetic similarity, 3 main clusters were distinguished, in which differentiation or a tendency towards differentiation by ecological-geographical groups was observed.

DNA typing of the gastric mucosa of patients with chronic duodenal ulcer

The work involved a molecular biological technique (ISSR-PCR) using ISSR-primer S-2, with structure (AGC) 6G. Changes in the gastric mucosa in chronic duodenal ulcer disease against the background of severe chronic atrophic gastritis have been analyzed. Noteworthy is the fact that there is a strong correlation between the degree of dysplasia of the epithelium of the gastric mucosa and the mitotic index, the Pearson's correlation coefficient rxy was 0.853, respectively. A strong and very strong correlation relationship between the indicators of the degree of dysplasia of the epithelium of the gastric mucosa by phenotypic characteristics and indicators of DNA typing of samples of the gastric mucosa, the Pearson's correlation coefficient rxy was 0.863, respectively. DNA profiles of the gastric mucosa of patients with duodenal ulcer according to the results of typing by the ISSR-PCR method ranging from 520 to 620 bp. had the character of microsatellite expansions and differed from the profile of the norm, which is evidence of precancerous changes.

Combination of Multiple Microsatellite Analysis and Genome-Wide SNP Genotyping Helps to Solve Wildlife Crime: A Case Study of Poaching of a Caucasian tur (Capra caucasica) in Russian Mountain National Park

Poaching is one of the major types of wildlife crime in Russia. Remnants of goats (presumably the wild endemic species, the Caucasian tur) were found in an area of the Caucasian mountains. The case study involves a suspected poacher whose vehicle was found to have two duffel bags containing pieces of a carcass, which he claimed was that of a goat from his flock. The aim of the forensic genetic analysis for this case was to (i) establish individual identity and (ii) perform species identification. DNA typing based on fourteen microsatellites revealed that STR-genotypes generated from pieces of evidence found at crime scene fully matched those obtained from the evidence seized from the suspect. The results of genome-wide SNP-genotyping, using Illumina Goat SNP50 BeadChip, provided evidence that the poached animal was a wild Caucasian tur (Capra caucasica). Thus, based on comprehensive molecular genetic analysis, evidence of poaching was obtained and sent to local authorities. To our knowledge, this case study is the first to attempt to use DNA chips in wildlife forensics of ungulates.

Probabilistic Genotyping of Single Cell Replicates from Complex DNA Mixtures Recovers Higher Contributor LRs than Standard Analysis

DNA mixtures are a common source of crime scene evidence and are often one of the more difficult sources of biological evidence to interpret. With the implementation of probabilistic genotyping (PG), mixture analysis has been revolutionized allowing previously unresolvable mixed profiles to be analyzed and probative genotype information from contributors to be recovered. However, due to allele overlap, artifacts, or low-level minor contributors, genotype information loss inevitably occurs. In order to reduce the potential loss of significant DNA information from donors in complex mixtures, an alternative approach is to physically separate individual cells from mixtures prior to performing DNA typing thus obtaining single source profiles from contributors. In the present work, a simplified micro-manipulation technique combined with enhanced single-cell DNA typing was used to collect one or few cells, referred to as direct single-cell subsampling (DSCS). Using this approach, single and 2-cell subsamples were collected from 2-6 person mixtures. Single-cell subsamples resulted in single source DNA profiles while the 2-cell subsamples returned either single source DNA profiles or new mini-mixtures that are less complex than the original mixture due to the presence of fewer contributors. PG (STRmixTM) was implemented, after appropriate validation, to analyze the original bulk mixtures, single source cell subsamples, and the 2-cell mini mixture subsamples from the original 2-6-person mixtures. PG further allowed replicate analysis to be employed which, in many instances, resulted in a significant gain of genotype information such that the returned donor likelihood ratios (LRs) were comparable to that seen in their single source reference profiles (i.e., the reciprocal of their random match probabilities). In every mixture, the DSCS approach gave improved results for each donor compared to standard bulk mixture analysis. With the 5- and 6- person complex mixtures, DSCS recovered highly probative LRs (> 1020) from donors that had returned non-probative LRs (<103) by standard methods.

Keeping race at bay: familial DNA research, the ‘Turkish Community,’ and the pragmatics of multiple collectives in investigative practice

In this contribution, we analyze the recently adjudicated Milica van Doorn rape and murder case. In this case, committed in 1992, no suspect could be identified until investigatory actors employed familial DNA searching in 2017. Crucially, familial DNA typing raised the possibility of ethnic and racial stereotyping and profiling, particularly against the background of the first case in which familial DNA typing was used in the Netherlands: the Marianne Vaatstra case, which from the start had been marred by controversy about the ethnicity of the unknown perpetrator. In our analysis, we show how criminal justice actors managed this potential for racialization through strategically mobilizing and carefully managing multiple collectives. Drawing on the notions of multiplicity and non-coherence, we show we do not only empirically trace the situated ethics and pragmatics of familial DNA research in this specific case, but we also develop a theoretical argument on the multiple and non-coherent character of race itself and its attendant ethical, political, and methodological possibilities and obligations.

Twenty-Seven Y-Chromosome Short Tandem Repeats Analysis of Italian Mummies of the 16th and 18th Centuries: An Interdisciplinary Research

Roccapelago (MO) is a small village located in the Northern Central Apennines, with a population of 31 inhabitants (2014). In 2010, more than 400 individuals dated between the end of the 16th and the 18th century, many of which partially mummified, were discovered in the crypt of the church. This small village, because of its geographical location and surrounding environment, seems to possess the characteristics of a genetic isolate, useful for population genetics and genealogical analyses. Thus, a diachronic study of DNA aimed at investigating the structure and dynamics of the population of Roccapelago over the about 4 centuries, was conducted by analyzing ancient and modern inhabitants of the village. The 14 modern samples were selected by considering both the founder surnames of the village, identified thanks to the study of parish registers, and the grandparent’s criterion. From 25 ancient mummies, morphologically assigned to male individuals, the petrous bone, that harbors high DNA amounts, was selected for the DNA extraction. The quantification and qualitative assessment of total human male DNA were evaluated by a real-time PCR assay using the Quantifiler Trio DNA Quantification Kit and multiplex PCR of 27 Y-chromosome short tandem repeat (Y-STR) markers included in the Yfiler Plus PCR Amplification Kit, with seven rapidly mutating Y-STR loci for improving discrimination of male lineages, was performed to genotype the samples. Y-STRs were analyzed according to the criteria of ancient DNA (aDNA) analysis to ensure that authentic DNA typing results were obtained from these ancient samples. The molecular analysis showed the usefulness of the Y chromosome to identify historically relevant remains and discover patterns of relatedness in communities moving from anthropology to genetic genealogy and forensics.

Reverse complement-PCR, an innovative and effective method for multiplexing forensically relevant single nucleotide polymorphism marker systems

DNA analyses from challenging samples such as touch evidence, hairs and skeletal remains push the limits of the current forensic DNA typing technologies. Reverse complement PCR (RC-PCR) is a novel, single-step PCR target enrichment method adapted to amplify degraded DNA. The sample preparation process involves a limited number of steps, decreasing the labor required for library preparation and reducing the possibility of contamination due to less sample manipulation. These features of the RC-PCR make the technology a unique application to successfully target single nucleotide polymorphisms (SNPs) in fragmented and low copy number DNA and yield results from samples in which no or limited data are obtained with standard DNA typing methods. The developed RC-PCR short amplicon 85 SNP-plex panel is a substantial improvement over the previously reported 27-plex RC-PCR multiplex that will provide higher discrimination power for challenging DNA sample analyses.