Monitoring of microbial communities of surface soil

Currently, the methods used for sampling soil do not meet modern requirements when conducting comprehensive studies of ecological state of soils. Stratified sampling of soil samples does not allow us to judge the gradients of distribution of agrochemical parameters and microbial communities of surface soil layer in natural landscapes. The use of method of GPS positioning of equidistant points of surface layer of soil in a comprehensive study of agrochemical parameters of soil, the composition of microbial communities of landscape makes it possible to draw up a summary map-scheme (GIS). The article analyzes methodological features of selection of soil samples for agrochemical, bacteriological, and molecular biological analysis. Using example of southern alkaline chernozem with gradient of pH distribution from 7.5 to 8.5 units, the distribution of microbial communities and agrochemical parameters in surface layer of soil of a perennial fallow is shown. The range of changes in agrochemical indicators on site with an area of 1 ha for organic matter was (1.4 ± 0.3% - 5.4 ± 0.5%), ammonium (12.3 ± 1.2 - 26.7 ± 2.7 mg/kg), and nitrate nitrogen (0.9 ± 0.1 - 3.9 ± 0.6 mg/kg). The change in microbial communities corresponded to gradient of soil agrochemical parameters.

Pengembangan Media Interaktif Mata Kuliah Teknik Analisis Biologi Molekuler (TABM)

Molecular Biology Analisis Techniques (TABM) is one of the courses that discusses the basic techniques needed for molecular biology. Learning in TABM, is more emphasis on procedural, so the process by using books alone is not enough. Therefore, to support the TABM learning process, interactive were developed consisting of text, sound, images, and research videos. This type of research is R & D is a research and development design by modifying the systematics of Kemp and Dayton. The development steps are to determine the specific learning objectives of the product; prepare material and video content; creating storyboard, developing, editing, and mxing; testing and revising. The instruments used are validation sheets and questionnaires for students with qualitative and quantitative data analysis techniques. Based on the result, there are several suggestions and inputs including: design, instructions for use, product packaging, introduction of tools and initial materials, and procedures. While quantitatively it can be seen that the developed interactive media doesn’t need to be revised and is suitable for use, namely 92,1% of field experts and 70,2% of media experts. It can be concluded that the development of interactive media can be used ini procedural courses such as molecular biological analysis techniques.

Transmembrane Protein Ttyh1 Maintains the Quiescence of Neural Stem Cells Through Ca2+/NFATc3 Signaling

The quiescence, activation, and subsequent neurogenesis of neural stem cells (NSCs) play essential roles in the physiological homeostasis and pathological repair of the central nervous system. Previous studies indicate that transmembrane protein Ttyh1 is required for the stemness of NSCs, whereas the exact functions in vivo and precise mechanisms are still waiting to be elucidated. By constructing Ttyh1-promoter driven reporter mice, we determined the specific expression of Ttyh1 in quiescent NSCs and niche astrocytes. Further evaluations on Ttyh1 knockout mice revealed that Ttyh1 ablation leads to activated neurogenesis and enhanced spatial learning and memory in adult mice (6–8 weeks). Correspondingly, Ttyh1 deficiency results in accelerated exhaustion of NSC pool and impaired neurogenesis in aged mice (12 months). By RNA-sequencing, bioinformatics and molecular biological analysis, we found that Ttyh1 is involved in the regulation of calcium signaling in NSCs, and transcription factor NFATc3 is a critical effector in quiescence versus cell cycle entry regulated by Ttyh1. Our research uncovered new endogenous mechanisms that regulate quiescence versus activation of NSCs, therefore provide novel targets for the intervention to activate quiescent NSCs to participate in injury repair during pathology and aging.

First cases of Hb Lepore in the Russian Federation

Hb Lepore, as the result of fusion of the b- and d-globin genes, leads to decreased amount of non-a-globin chains availability for hemoglobin formation. Hb Lepore, up to now, was not identified among Russian patients. We provide clinical and laboratory information on Hb Lepore Boston–Washington in two cases, one of them familial. A small amount of abnormal Hb was detected by capillary electrophoresis, an abnormal globin chain was shown by HPLC, and the final diagnosis of Hb Lepore Boston– Washington was made by molecular biological analysis of globin genes. Peripheral blood for all affected people revealed RBC’s hypochromia microcytosis and normal Hb concentration. The parents of the patients agreed to use the information, including photos of children, in scientific research and publications.

Comparison of Epidermal Barrier Integrity in Adults with Classic Atopic Dermatitis, Atopic Prurigo and Non-Atopic Prurigo Nodularis

A deficient epidermal barrier is a key feature of atopic dermatitis (AD) and comprises altered lipid and protein content and composition of the stratum corneum resulting in disturbed water balance. Clinically, eczematous lesions on dry skin and pruritus develop. Pruritic nodules occur in prurigo nodularis (PN), another chronic skin disease, which can be associated with atopy. We aimed at comparing the three clinical pictures, classic AD, atopic prurigo (AP), and non-atopic PN, to healthy controls regarding the epidermal barrier. We determined clinical parameters and performed biophysical measurements, histology/immunohistochemistry, electron microscopy, and molecular biological analysis. We found distinctively elevated clinical scores, reduced hydration and increased transepidermal water loss, epidermal hyperplasia and inflammation reduced filaggrin and increased loricrin and involucrin expression, as well as reduced intercellular lipid lamellae in all three disease groups. These findings show a severe disruption in epidermal barrier structure and function in all three disorders so that epidermal barrier impairment is now proven not only for AD but also for PN.

Fermented Brown Rice as a Functional Food

Brown rice, especially in a part of rice bran, contains many kinds of nutrients and biologically active components such as plant polyphenols and phytic acid, but is hard to eat. “Brown rice and rice bran fermented with Aspergillus oryzae (FBRA)” is a processed food that is easier for daily intake, commercially available, and rich in eating experience. During the fermentation process, dietary fibers is partially digested, and free vitamins and phenolic compounds have increased. These fermentation products are utilized for quality control to manage FBRA production. Recently, plant-derived polyphenols have shown anti-oxidative activity and biological function in various disease models. We and other research groups used raw powder FBRA to examine its biological activity through pathological and/or molecular biological analysis. Dietary administration of FBRA showed anti-tumorigenic effects in chemically induced tumors in rodents. Anti-inflammatory effects have been observed in DSS-induced colitis in rat and inflammation-mediated rodent tumor models. I will give an outline of the characteristic of FBRA, and then introduce our recently published work about “Preventive effect of FBRA on spontaneous type 1 diabetes in NOD female mice”, including how to estimate the in vivo effect of dietary FBRA, its possible mechanisms and the limit of this study.

P–201 The beneficial effects of ZP-free culture on cytoplasmic fragmentation in human embryos. : An innovative trial using 3PN zygotes

Study question Is it possible to culture ZP-free embryos to eliminate perivitelline threads, which are known to be involved in generating cytoplasmic fragments at the first cleavage? Summary answer ZP-free culturing, an innovative system that decreases the amount of cytoplasmic fragments without disrupting the blastomeres, using incubators with time-lapse imaging. What is known already A study in 2017 observed perivitelline threads in more than 50% of cleavage-stage human embryos using time-lapse imaging, and the rate of cytoplasmic fragmentation (at the first cleavage) was significantly decreased in embryos without perivitelline threads (P < 0.001). While it has been proposed that perivitelline threads play an important role in crosslinking the cumulus cells and oocyte during maturation, the mechanism underlying such a role remains unclear. It is also unknown whether the threads still function in mature MII oocytes. Study design, size, duration A prospective study was conducted using 2,852 normal (2PN/2PB) embryos from c-IVF/ICSI and 113 abnormal (3PN) embryos obtained from c-IVF between 2017 and 2019. The zona pellucida (ZP) of 71 abnormal embryos was removed at the pronuclear stage (“ZP-free”), and the rest (n = 42) were cultured as “ZP-intact”. Normal and abnormal embryos were cultured for five days in bench-top incubators (MINC, COOK) and an incubator equipped with a time-lapse imaging system. Participants/materials, setting, methods Embryos used in this study were donated by 412 couples who underwent c-IVF cycles in our clinic between 2017 and 2019. For ZP removal, 3PN embryos were placed in 0.125M sucrose-containing HEPES media drops to reduce the ooplasm size. Then, ooplasms were completely separated from ZPs by a laser and pipetting. Embryo development and morphology of the three groups (normal, ZP-intact and ZP-free abnormal) were compared based on the degree of cytoplasmic fragmentation. Main results and the role of chance The first cleavage occurred in 97.8% (n = 2,790/2,852) of 2PN/2PB, 83.3% (n = 35/42) of ZP-intact 3PN and 97.2% (n = 69/71) of ZP-free 3PN. Normal (2PN/2PB), ZP-intact and ZP-free 3PN embryos were classified into three groups based on the modified Veeck’s criteria thus: <20% fragmented compared to the total volume of cytoplasm at the first cleavage (Grade 1 and 2, Good); 20–39% fragmented (Grade 3, Fair) and ≧40% fragmented (Grade 4, Poor). Of 69 cleaved ZP-free 3PN embryos, 68.1% (n = 47) showed less than 20% fragments which was significantly higher than 2PN/2PB (43.7%, n = 1,218/2,790) and ZP-intact 3PN (45.7%, n = 16/35; P < 0.05). Furthermore, 24.6% (n = 17/69) of ZP-free 3PN embryos showed 20–39% fragments which was significantly lower than 2PN/2PB (45.9%, n = 1,281/2,790; P < 0.05). In addition, 50.7% of ZP-free 3PN embryos (n = 36) developed to the morula stage after the third cleavage, and 29.6% (n = 21) formed blastocoel and became blastocysts. Thus, removing the ZP before the first cleavage did not adversely affect embryo development and decreased the cytoplasmic fragmentation. Limitations, reasons for caution Due to ethical and clinical limitations, we only examined abnormally fertilized embryos in this study. Moreover, since the relationship between the perivitelline threads and cytoplasmic fragments is unclear, we plan to conduct molecular biological analysis of the perivitelline threads in further studies. Wider implications of the findings: This study revealed that ZP is not always necessary after the pronuclear stage because ZP-free embryos studied herein developed normally and maintained cell adhesion well. This innovative culture method might provide the breakthrough needed for patients to improve embryo quality who obtain embryos with severe fragmentation caused by perivitelline threads. Trial registration number Not applicable

Analysis of dairy productivity of cows of ukrainian brown dairy breed of different genotypes by kappa-casein

In the herd of cattle of the Ukrainian Brown Dairy breed studies were conducted, the purpose of which was to study the influence of the genotype of cows by kappa-casein on the indicators of their milk productivity. Genotyping of 29 heads of cattle. Determination of the polymorphism of the kappa-casein gene was performed in the genetic laboratory of the Institute of Physiology n.a. Bogomolets of NAS using molecular biological analysis of allele recognition by polymerase chain reaction (PCR) in real time. Allele A had a slightly higher frequency - 0.517 compared to allele B - 0.483 at the locus of the kappa-casein gene, although it was not significant. As a result, the proportions of AA and BB genotypes were high (38 and 34%, respectively). The performed genetic-statistical analysis revealed an excess in the kappa-casein locus of homozygous variants AA and BB and a lack of heterozygous AB. The degree of homozygosity, which is 50.1%, indicates a sufficient level of consolidation in the studied population of cattle. The number of effective active alleles in the kappa-casein locus of cows of the Ukrainian Brown Dairy breed is 1.99, with the maximum value of the level of polymorphism possible at a two-allele locus of 2. A negative heterozygosity test (HT) indicates a lower proportion of actual heterozygotes relative to the proportion of theoretical heterozygotes. The reason for the existing genetic structure of the herd of Ukrainian Brown Dairy breeds by the kappa-casein locus was the use of Swiss cattle breeders, most of which were not evaluated by the genotype of the kappa-casein gene. Studies of the level of milk productivity of cows of different genotypes by kappa-casein in the first lactation did not reveal a significant difference. At the same time, heterozygous (AB) animals were inferior to homozygous (AA and BB) in terms of milk yield, and animals with AA and AB genotypes were preferred in terms of quality. For the best lactation, a significant difference was found in the amount of milk yield and the amount of milk fat. Homozygous animals (AA and BB) significantly (p <0.05) outperformed heterozygous (AB) in milk yield, by 1091 and 922 kg, respectively. Animals with heterozygous AB genotype in terms of the average amount of milk fat were significantly inferior to homozygous AA and BB animals (p <0.05). The small number of experimental animals was one of the reasons for the discrepancy between the results of research and the data of other scientists. Key words: breed, milk yield, fat content, protein content, kappa casein, genotype, allele.

Genetic Diversity of Natural Morchella spp. from the Southern Gansu Province Based on rDNA-ITS, China

Background: China is a mainland country rich in natural Morchella spp. resources. At present, about half of the natural Morchella spp. in the world has been recorded in China. The current study was aimed at the classification and cultivation of Morchella spp. This study provided a more accurate molecular trait for the systematic classification of Morchella spp. in southern Gansu Province, China.Methods: From April to May 2019. Based on the molecular biological analysis of 16 natural Morchella spp. strains collected from the southern Gansu province, China. the ITS fragments between transcripts of the rDNA gene were amplified by PCR and sequenced.Result: The results of BLAST comparison according to GenBank showed that the 16 tested strains of Morchella spp. were classified into 5 species, which were M.angusticeps, M.esculenta, M.elata, M.crapssipes and M.conica. According to the molecular evolutionary trees constructed by the Maximum-Parsimony method (MP) and the Neighbour-Joining method (NJ), the topological structures of the two molecular evolutionary trees were similar and the Bootstrap verification had a high support rate, indicating that the phylogenetic relationship had high credibility.