Laboratory diagnostics of Blastocystis infections – challenges and controversies

Blastocystis is a unicellular intestinal organism with undetermined pathogenicity, one of the most commonly detected in human stool samples around the world. Sometimes their presence may contribute to the development of blastocystosis manifested by gastrointestinal complaints such as abdominal pain, diarrhea, flatulence, and nausea of varying severity with periods of exacerbation and remission. Due to the significant polymorphism (at least 4 morphological forms and a large range of sizes), and the delicate, easily degradable structure, recognition of Blastocystis in microscopic stool smears is difficult. They are easily overlooked or confused with other microorganisms present in the feces. There is a lack of a standardized procedure for the diagnostics of blastocystosis. In inter-laboratory comparative studies, Blastocystis is the most inconsistently detected intestinal eukaryote. The article discusses the difficulties in diagnosing blastocystosis, and the methods used to detect Blastocystis in terms of their usefulness in a medical diagnostic laboratory.

PCR- RFLP Polymorphism at Exon 3 and Promoter Region of Prolactin Gene in Gaolao Cattle

The present study was planned to investigate significant polymorphism in exon3 and promoter regions of prolactin gene in Gaolao cattle and its association (if any) with milk components. A total of 50 randomly selected Gaolao cows were genotyped for PRLGI-RsaI and PRLGIII-Tsp45I loci using PCR-RFLP technique. The PRLGI-RsaI locus was polymorphic and PRLGIII-Tsp45I locus was monomorphic indicating genetic variation at PRLGI-RsaI locus in Gaolao cattle. The allele frequency for the PRLA alleles was found 0.933 and for allele PRLB was 0.067 at PRLGI–RsaI locus. The result of analysis variance showed significant (PÂ0.05) effect of AA genotype on fat % as compared to AB genotype; however non-significant effect of these genotypes was found on protein %, SNF % and lactose % which may be due to small sample resulting in higher error variance. The results of present study indicated that identified prolactin gene polymorphism at PRLGI-RsaI in exon 3 after validation in large breeding populations having precise data records may prove as important candidate gene marker for selection and breeding decisions in genetic improvement of indigenous cattle for milk productivity.

CREMATION AT MEDIEVAL SITES OF THE VETLUGA REGION: BIOARCHAEOLOGICAL ASPECTS OF RESEARCH

Важную составляющую в характеристике погребальной обрядности формируют сведения об останках погребенного. Кремированные останки не доносят до нас привычной для палеоантропологического исследования информации, тем не менее важны для характеристики обрядовых действий. Объектом нашего исследования стала погребальная практика средневековых финно-угорских племен Поветлужья VI–XII вв. Нами привлечены материалы из могильников: Черемисское кладбище, Выжумский, Веселовский и Кузинские хутора из раскопок Т. Б. Никитиной 2008–2014 гг. Археологические характеристики позволяют выделить две группы памятников. Формальные характеристики состояния кремированных останков не позволяют проводить четкую дифференциацию. Выражен значительный полиморфизм погребальной обрядности. В связи с этим возникает предположение, что особенности погребения обусловлены не столько локальной принадлежностью, сколько личностью погребенного и обстоятельствами его смерти. Описанные материалы, хотя и являются немногочисленными, но позволили реконструировать спектр вариантов погребальной церемонии средневековых марийцев X–XI вв. Подчеркнем, что по ряду параметров скопления кремаций из двух марийский могильников X–XI вв. (2-я группа) напоминают скопления кремированных костей из погребений средневековой муромы Подболотьевского могильника. Information on human fossil remnants is an important component characterizing funerary rites. Cremated remains do not contain information that is usually used in a paleoanthropological study; nevertheless, they are important for characterizing ritual actions. This study is focused on the funerary practice of medieval Finno-Ugric populations inhabiting the Vetlugaregion in the 6th – 12th centuries. For our research we used materials from a number of cemeteries excavated by T. B. Nikitina in 2008–2014: the Cheremisskoye cemetery, the Vyzhumskiy, the Veselevolskiy and the Kuzinskiye farmsteads. Based on archaeological characteristics, two groups of sites can be singled out. Formal characteristics of conditions of cremated remnants do not provide an opportunity to have clear differentiation. Significant polymorphism of funerary practices is quite evident. All this may imply that specific features of the grave are stipulated by a personality of the deceased and circumstances of his/her death rather than local attribution. The materials described, though few in number, enabled us to reconstruct a range of funerary rite variants practiced by medieval Mari people in the 10th – 11th centuries. We would like to emphasize that, regarding a number of characteristics, concentrations of cremations from two Mari cemeteries of the 10th – 11th centuries (group 2) resemble concentrations of cremated bones from the medieval Muroma graves in the Podbolot’yevo cemetery.

Characteristic of the zimoseptoria tritici population by virulence in the of the Central Black Soil region of Russia

Based on the analysis of the intraspecific structure of the fungus Zimoseptoria. tritici, it was found that this species has significant polymorphism. Using monogenic wheat lines (Oasis (Stb1), Veranopolis (Stb2), Israel (Stb3), Tadinia (Stb4), CS / Synthetic (Stb5), Estanzuela Federal (Stb7)), the population of Z. tritici in the Central Black Earth region was tested. High heterogeneity of monosporous isolates of Z. tritici by virulence was revealed.

ПОЛИМОРФНЫЕ ВАРИАНТЫ ГЕНОВ МАТРИКСНЫХ МЕТАЛЛОПРОТЕИНАЗ У ДЕТЕЙ С АТОПИЧЕСКИМ ДЕРМАТИТОМ

Обоснование. Матриксные металлопротеиназы (MMP) играют особую роль в патогенезе атопического дерматита (АтД). Поэтому исследование особенностей наследования генов, отвечающих за синтез ММП у детей, страдающих АтД, представляет большой научный и практический интерес. Цель. Изучить роль полиморфных вариантов генов матриксных металлопротеиназ (ММР9 и ММР20) в патогенезе АтД у детей. Материалы и методы. Методом аллельспецифичной полимеразной цепной реакции проведено исследование аллельных вариантов 320AC гена MMP20, 837C гена MMP20, 8202 AG гена MMP9 у детей с АтД. Контрольную группу составили пациенты I и 11а групп здоровья соответствующего пола и возраста. Результаты. Результаты проведенных генетических исследований показали, что частота встречаемости аллелей и генотипов по полиморфизмам 320AC гена MMP20,837rYC гена MMP20 среди больных не имела достоверных отличий от группы контроля (р0,05). При изучении полиморфизма 8202 AG гена MMP9 установлено, что среди детей, страдающих АтД, преобладает А/Агенотип с частотой 69,2, тогда как в группе здоровых детей частота данного генотипа в 3 раза ниже (р0,003). В группе контроля преобладающим является A/Gгенотип (55,7), а носителей G/Gгенотипа в 2 раза меньше (21,3). Важно отметить, что у детей с генотипом А/А риск развития АтД повышен в 7,55 раза (OR7,55 95 Cl 2,9719,21 р0,001. Заключение. Таким образом, можно предположить, что наиболее значимым полиморфизмом в патогенезе АтД у детей является 8202AG гена MMP9, а именно у гомозигот по Ааллели риск развития кожных проявлений аллергии повышен более чем в 7 раз.Matrix metalloproteinases (MMP) play a special role in the pathogenesis of atopic dermatitis (AD). Therefore, the study of the features of genes responsible for the synthesis of MMP in children with AD is of great scientific and practical interest. Background. To study the role of polymorphic variants of matrix metalloproteinase genes (MMP 9 and MMP20) in the pathogenesis of AD in children. Materials and methods. Allelic variants of320AC gene MMP20,837TC gene MMP20, 8202 AG gene MMP9 in children with AD were studied using the method of allelespecific polymerase chain reaction. The control group consisted of I and IIa the health groups patients of the corresponding sex and age. Results. The results of genetic studies showed that the incidence of alleles and genotypes in the polymorphisms 320AC of the gene MMP20, 837TC of the gene MMP20 in patients had no significant differences from the control group (p0.05). It was established that the A/Agenotype of polymorphism 8202 AG of the MMP9 gene, prevailed among children suffering from AD at a frequency of 69.2, whereas in the group of healthy children the frequency of this genotype was 3 times lower (p0.003). At the same time A/Ggenotype (55.7) was predominant in the control group, while G/G genotype was 2 times lower (21.3). Thus the risk of AD increased by 7.55 times (OR7.55 95 Cl 2.9719.21 p0.001 in children with genotype A/A. Conclusion. The most significant polymorphism in the pathogenesis of AD in children is 8202 AG of the MMP9 gene, in particular the risk of developing of skin manifestations of allergy is increased by more than 7 times in Aallele homozygotes.

Features of the prevalence of TLR-2 gene polymorphism (Arg753Gln) in patients with acute brucellosis with liver damage

Brucella have the potential influence for all organs of the human body, which is reflected by a significant polymorphism of clinical manifestations, as a result, not always timely diagnosis of the disease and the development of complications that result of disability. The aim of our study was to determine the characteristics of the prevalence of polymorphism (Arg753Gln) of the TLR-2 gene in patients with acute brucellosis with liver damage. 178 patients with clinical symptoms of brucellosis were examined. According to the criteria for inclusion in a study of 178 patients, only 120 persons, who were the main group, fully met all criteria. All patients were tested for polymorphism (Arg753Gln) TLR-2. Statistical processing the results of the study was performed using the programs “SPSS 20.0”, “STATISTICA 6.0”. Among 120 patients with acute brucellosis, the disease of the hepatobiliary system had only 43 patients, which was 35.8% according to the laboratory and instrumental methods of investigation. The Gln/Gln genotype of the TLR-2 gene was determined in 3.3 times more in patients with acute liver brucellosis than in the patients without liver damage. Among the carriers of the genotype Gln/Gln, there is a significantly increased risk of brucellosis (χ2=17.52; p=0.0002; OR=4.81; 95% CI [0.98-23.59]), while carries of the homozygous genotype Arg/Arg on the contrary, has a protective effect on the development of brucellosis (OR=0.11, 95% CI [0.04 - 0.33]).

Vaginal biocenosis state assessment in patients with cervical ectopy relapse in the conditions of human papillomavirus infection

Cervical ectopy is one of the most common benign processes, characterized by relatively high recurrence rate, and in the case of vaginal microbiota violations in the conditions of HPV infection combination deserves the aimed attention, particularly in the context of reproductive health care and cancer prevention. The objective: to study the vaginal microbiota features in patients withcervical ectopy relapsein the conditions of HPV infection. Materials and methods. The study included 98 women of reproductive age (18 to 46 years). Three groups were formed: a control group (n=30), in which conditionally healthy women were included, group 1 (n=33) – patients with firstly diagnosed cervical ectopy, group 2 (n=35) – patients with cervical ectopy relapse. All women wereexamined with a bacterioscopic, cytological, microbiological study of the vaginal biocenosis, as well as a determination of the vaginal environmentpH level. Results. In patients with cervical ectopy relapse the pH of the vaginal environmentwas significantly higher than that in women of the control group (p<0.001) and of the group 1 patients (p<0.05). About 9% of patients in both study groups had signs of bacterial vaginosis; in about 70% of group 2 patients the signs of mild and moderate severity aerobic vaginitis were found, significantly (p<0.05) more often than in patients with firstly diagnosed cervical ectopy. In group 2 patients a significant polymorphism of infects was detected in 40.00±8.28% of cases, the associations of 3 or more microorganisms determined simultaneously were significantly (p<0.001) more often foundin group 2 in comparison with patients with firstly diagnosed cervical ectopy. Сonclusion. The use of the short-term test system A.F.GENITAL SYSTEM (Liofilchem®, Italy) has a number of advantages for clinical practice: results being ready in short time, the detection of a wide range of infections, the possibility of quantifying M. hominis and U. urealyticum, and then determining the sensitivity to the most frequently used antibacterial drugs. Recurrent cervical ectopy often occurs in the conditions of colpitis, vaginitis, vaginal microbiota disorders with significant polymorphism of microorganisms and alkalization of the vaginal environment, which complicates the processes of ectopy epithelialization and can predispose to its recurrence. The observed cytological features in this cohort of patients suggest that long-term violations of the vaginal biocenosis in combination with the inflammatory processes of the vagina and cervix create favorable conditions for the realization of dysplastic changes caused by HPV. When determining the tactics of treatment of recurrent cervical ectopy, in particular in the conditions of HPV infection, first of all it is necessary to achieve both the cytological norm and the normalization of the vaginal biocenosis before any intervention, if necessary. Key words: relapse of cervical ectopy, vaginal biocenosis, vaginitis, cervicitis, HPV.

Identification and comparison of individual chromosomes of three accessions of Hordeum chilense, Hordeum vulgare, and Triticum aestivum by FISH

Karyotypes of three accessions of Hordeum chilense (H1, H16, and H7), Hordeum vulgare, and Triticum aestivum were characterized by physical mapping of several repetitive sequences. A total of 14 repetitive sequences were used as probes for fluorescence in situ hybridization (FISH) with the aim of identifying inter- and intraspecies polymorphisms. The (AG)12 and 4P6 probes only produced hybridization signals in wheat, the BAC7 probe only hybridized to the centromeric region of H. vulgare, and the pSc119.2 probe hybridized to both wheat and H. chilense, but not to H. vulgare. The remaining repetitive sequences used in this study produced a hybridization signal in all the genotypes. Probes pAs1, pTa-535, pTa71, CCS1, and CRW were much conserved, showing no significant polymorphism among the genotypes studied. Probes GAA, (AAC)5, (CTA)5, HvT01, and pTa794 produced the most different hybridization pattern. We identified large polymorphisms in the three accessions of H. chilense studied, supporting the proposal of the existence of different groups inside species of H. chilense. The set of probes described in this work allowed the identification of every single chromosome in all three species, providing a complete cytogenetic karyotype of H. chilense, H. vulgare, and T. aestivum chromosomes, which could be useful in wheat and tritordeum breeding programs.

Identification and comparison of individual chromosomes of three Hordeum chilense accessions, Hordeum vulgare and Triticum aestivum by FISH

Karyotypes of three accessions of Hordeum chilense (H1, H16 and H7), Hordeum vulgare and Triticum aestivum were characterized by physical mapping of several repetitive sequences. A total of fourteen repetitive sequences were used as probes for fluorescence in situ hybridization (FISH) with the aim of identifying inter‐ and intra-species polymorphisms. The (AG)12 and 4P6 probes only produced hybridization signals in wheat, the BAC7 probe only hybridized to the centromeric region of H. vulgare, and the pSc119.2 probe hybridized to both wheat and H. chilense, but not to H. vulgare. The remaining repetitive sequences used in this study produced a hybridization signal in all the genotypes. Probes pAs1, pTa535, pTa71, CCS1 and CRW were much conserved, showing no significant polymorphism among the genotypes studied. Probes GAA, (AAC)5, (CTA)5, HvT01 and pTa794 produced the most different hybridization pattern. We identified large polymorphisms in the three accessions of H. chilense studied, supporting the proposal of the existence of different groups inside H. chilense species. The set of probes described in this work allowed the identification of every single chromosome in all three species, providing a complete cytogenetic karyotype of H. chilense, H. vulgare and T. aestivum chromosomes, useful in wheat and tritordeum breeding programs.

ВИЗНАЧЕННЯ ОСОБЛИВОСТЕЙ ГЕНОТИПІВ У ЖІНОК З ГЕНІТАЛЬНИМ ЕНДОМЕТРІОЗОМ ТА ФІБРОЗНО-КІСТОЗНОЮ МАСТОПАТІЄЮ ТА ЇХ РОЛЬ У ТЯЖКОСТІ ПЕРЕБІГУ ЗАХВОРЮВАННЯ

The scientific work is devoted to the urgent problem of our time - dishormonal disease of the female reproductive system, namely endometriosis and fibrosis-cystic mastitis. During the last years there has been a steady increase in the dual pathology of genital and breast diseases. According to statistics dishormonal frequency of breast diseases in the population of the female population suffering from various gynecological diseases varies between 76-98 %. It is known that the risk of this disease is closely associated with some alleles detoxification system, glutathione family - S- transferase and proonkogenom p 53, characterized by significant polymorphism. The article presents the results of research on the role of deletion variants of genes GSTT1, GSTM1 and p 53 in the development of endometriosis and fibrosis-cystic mastitis. Screening populations on carriage of alleles reveals women who are likely to develop these diseases and choose the most effective tactics of medical treatment in the event of the disease.