Modification of the Method for Determining Myeloperoxidase in Blood Neutrophils

Myeloperoxidase is a heme-containing peroxidase expressed primarily in neutrophils and to a lesser extent in monocytes. Determining the activity of myeloperoxidase in blood cells is one of the tests of the immune status of animals. Conventional methods are based on the oxidation of benzidine by the peroxide system to the unstable benzidine blue, which spontaneously turns into stable brown benzidine. The aim of this study was to develop a modification of the cytological determination of the myeloperoxidase enzyme using metol. The relative percentage of peroxidase-positive neutrophils in the peripheral blood of animals was determined after 100 neutrophils had been counted. The task was achieved by using the reaction with metol in the method of cytological determination of the activity of neutrophil myeloperoxidase in animal blood smears, which was based on the oxidation of metol by a peroxide system. Images of micropreparations were digitized using a Sony device for processing the received images of the cells. The Image Tool computer program was used for this purpose. The biological substrate was processed from a buffer-incubation mixture with subsequent drying and microscopy. The main new modification of the method was using metol. Metol does not have the ability to inhibit the activity of myeloperoxidase. The research showed easy and fast results. This method is economical and perspective for using in practice. Keywords: myeloperoxidase, blood, neutrophils, meto

PECULIARITIES OF AN ASSOCIATION OF POLYMORPHOUS MARKERS -251 Т>А OF IL-8 GENE AND POLYMORPHISM OF HELICOBACTER PYLORI GENES IN NATIVE AND VISITANT POPULATION OF THE REPUBLIC OF KHAKASSIA

We researched, polymorphism, of interleukin IL-8 gene at Helicobacter pylori-associated diseases: ulcers and chronic gastritis in native and visitant population of the Republic of Khakassia by the method, of restriction enzyme digest analysis. Prevalence of subtypes VacA+ and CagA+ of Helicobacter pylori was realized, by the method, of polymerase chain reaction. We revealed, the most widespread allelic variants of genes of cytokines among Khakas. The expediency of determination, of population, risk and. protective genotypes of Helicobacter pylori that promote development of ulcer desease in Khakas

Localization of the quantitative trait loci related to lodging resistance in spring bread wheat (<i>Triticum aestivum</i> L.)

The yield and grain quality of spring and winter wheat significantly depends on varieties’ resistance to lodging, the genetic basis of this trait being quantitative and controlled by a large number of loci. Therefore, the study of the genetic architecture of the trait becomes necessary for the creation and improvement of modern wheat varieties. Here we present the results of localization of the genomic regions associated with resistance to lodging, plant height, and upper internode diameter in Russian bread wheat varieties. Phenotypic screening of 97 spring varieties and breeding lines was carried out in the field conditions of the West Siberian region during 2017–2019. It was found that 54 % of the varieties could be characterized as medium and highly resistant to lodging. At the same time, it was noted that the trait varied over the years. Twelve varieties showed a low level of resistance in all years of evaluation. Plant height-based grouping of the varieties showed that 19 samples belonged to semi-dwarfs (60–84 cm), and the rest were included in the group of standard-height plants (85–100 cm). Quantitative trait loci (QTL) mapping was performed by means of genome-wide association study (GWAS) using 9285 SNP markers. For lodging resistance, plant height, and upper internode diameter, 26 significant associations (–log p &gt; 3) were found in chromosomes 1B, 2A, 3A, 3D, 4A, 5A, 5B, 5D, 6A, and 7B. The results obtained suggest that the regions of 700–711 and 597–618 Mb in chromosomes 3A and 6A, respectively, may contain clusters of genes that affect lodging resistance and plant height. No chromosome regions colocalized with the QTLs associated with lodging resistance or upper internode diameter were found. The present GWAS results may be important for the development of approaches for creating lodging-resistant varieties through marker-assisted and genomic selection

Lodging in wheat: genetic and environmental factors and ways of overcoming

Lodging is one of the main factors in reducing the yield and grain quality of winter and spring wheat varieties. The resistance of wheat cultivars to lodging largely depends on environmental factors, biological and morphological features of the stem and root systems. Selection of the varieties for resistance to lodging is relevant in many countries of the world and has a number of achievements. Plant height is one of the most important morphological characters associated with lodging resistance. Breeding of the varieties carrying the dwarfing genes (Rht) is the main direction to reduce the risk of lodging. The Rht-B1b, Rht-D1b, Rht8 and Rht11 genes are widely used throughout the world due to their significant influence on agronomically valuable traits, including lodging. It turned out to be important to study the anatomical and morphological features and chemical composition of stem tissues, which complement the assessment of resistance to lodging and allow the varietal material to be more fully characterized. The thickness of stem internodes and their anatomical structure play an important role in the stem strength. The diameter of the stem, its thickness and weight, a large number of vascular bundles and a wide ring of mechanical tissues correlate with resistance to lodging. The content of lignin, silicon and cellulose are important structural components and provide the stem strength of wheat plants. Molecular genetic analysis and mapping of genes and quantitative trait loci are of great importance in identifying the genetic basis of the relationship between the anatomical and morphophysiological characters of the stem and root system and lodging. Genetic factors reflecting correlations between the lodging and the thickness of the stem wall, the number of vascular bundles and other characters were mapped to chromosomes 1A, 1B, 2A, 2D, 3A, 4B, 4D, 5A, 5D, 6D and 7D. It has been found that loci with high phenotypic effects on lodging tolerance are colocalized with loci responsible for plant height, stem diameter and stem strength. To increase resistance to lodging, it is necessary to develop a set of agrotechnical methods that reduce the influence of soil and climatic factors and create wheat varieties tolerant to lodging

Brown mycelial mat as an essential morphological structure of the shiitake medicinal mushroom lentinus edodes (Agaricomycetes)

© 2017 Begell House, Inc. We show here, to our knowledge for the first time, that the brown mycelial mat of the xylotrophic shiitake medicinal mushroom, Lentinus edodes, not only performs a protective function owing to significant changes in the ultrastructure (thickening of the cell wall, increased density, and pigmentation of the fungal hyphae) but also is a metabolically ac tive stage in the development of the mushroom. The cells of this morphological structure exhibit repeated activation of expression of the genes lcc4, tir, exp1, chi, and exg1, coding for laccase, tyrosinase, a specific transcription factor, chitinase, and glucanase, which are required for fungal growth and morphogenesis. This study revealed the maximum activity of functionally important proteins with phenol oxidase and lectin activities, and the emergence of additional laccases, tyrosinases, and lectins, which are typical of only this stage of morphogenesis and have a regulatory function in the development and formation of fruiting bodies

Состояние глазной поверхности при псевдоэксфолиативном синдроме

In the article the ocular surface was assessed in groups of patients with and without pseudoexfoliative syndrome. Tear secretion (Schirmer test II) and break up time reduction and significant damage of conjunctiva surface (staining with fluorescein) weremarked in group of patients with pseudoexfoliative syndrome. According to questionnaire which reflects index of damage of ocular surface, clinical implications of ocular surface disease was higher in group of patients with pseudoexfoliative syndrome.Проведена оценка состояния глазной поверхности в группах обследованных с псевдоэксфолиативным синдромом и без него. В группе с псевдоэксфолиативным синдромом отмечается статистически значимое снижение слезопродукции (тест Ширмера II), времени разрыва слезной пленки и выраженное повреждение поверхности конъюнктивы (прокрашивание флюоресцеином). По данным опросника, отражающего индекс повреждения глазной поверхности, у пациентов с псевдоэксфолиативным синдромом более выражены клинические проявления синдрома сухого глаза

СОСТОЯНИЕ ТКАНЕЙ ГЛАЗНОЙ ПОВЕРХНОСТИ У ПАЦИЕНТОВ С ИНВОЛЮЦИОННЫМИ НАРУШЕНИЯМИ ПОЛОЖЕНИЯ НИЖНЕГО ВЕКА

The article presents the results of the evaluation of the state of the ocular surface in patients with involutional lower eyelid malpositions of atopic type. In the group with eyelid malposition was marked a statistically significant reduction in the time gap of the tear film and marked damage of the surface of the cornea and conjunctiva. According to the questionnaire, the index reflecting the damage to the ocular surface tissues in patients with malposition eyelids more pronounced clinical manifestations of dry eye syndrome. Therefore, involutional lower eyelid malpositions lead to the defeat of the ocular surface tissue, mainly due to the violation of the tear film stability and enhance its evaporation.Приведены результаты оценки состояния глазной поверхности у пациентов с нарушением положения нижнего века атонического характера. В группе с мальпозициями век отмечается статистически значимое снижение времени разрыва слезной пленки и выраженное повреждение поверхности конъюнктивы и роговицы. По данным опросника, отражающего индекс повреждения тканей глазной поверхности, у пациентов с мальпозициями век более выражены клинические проявления синдрома сухого глаза. Таким образом, инволюционные нарушения положения нижнего века приводят к поражению тканей глазной поверхности, преимущественно за счет нарушения стабильности слезной пленки и повышения ее испарения

Melatonin in the dermal film limits the blood lymphocyte death in experimental thermal trauma

According to WHO data, about 11 million people need medical care after burns every year. In the overall structure of burns, the share of thermal trauma (TT) is 80%. Lymphocytopenia in TT is a risk factor for infectious complications and limited repair, and the development of new tools for TT therapy using dermal films is demanded in combustiology. The aim of the study was to evaluate changes in blood lymphocyte parameters, i.e., quantitative composition and their death during experimental thermal damage under the influence of the originally developed dermal film with melatonin (MT) in 49 inbred rats. The grade IIIA TT of 3.5% body surface was modeled by contact with boiling water for 12 s. Dermal films based on sodium carboxymethylcellulose supplemented with MT at a concentration of 0.005 g/g were applied daily for 5 days. The total numbers of lymphocytes, CD45RA+ and CD3+ cells, counts of lymphocytes with signs of partial necrosis, early and late apoptosis were assessed in blood. Relative decrease in the area and rate of the burn wound epithelization were also calculated. In animals with TT, the number of blood lymphocytes decreased on days 5, 10 and 20, including CD45RA+ and CD3+, along with increased amounts of lymphocytes with signs of necrosis, late and early apoptosis. By the term of 20 days, the burn wound area was reduced by 11.5%. Usage of dermal films with MT increased the amount of CD3+ cells in blood on days 5 and 10, CD45RA+ on days 5, 10 and 20, being associated with decreased number of lymphocytes showing signs of early apoptosis on days 5, 10 and 20, as well as features of necrosis and late apoptosis on days 5 following TT, accelerates the healing of a burn wound on days 5, 10 and 20 after TT. with a 20 cent reduction of its area by the day 20. Epithelization rate of the burn wound when applying MT-supplemented dermal film on days 5, 10 and 20 increases, along with higher amounts of CD3+ in the blood, and reduced counts of lymphocytes with signs of early apoptosis

Alteration in the ultrastructural morphology of mycelial hyphae and the dynamics of transcriptional activity of lytic enzyme genes during basidiomycete morphogenesis

© 2017, The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg.The morphogenesis of macromycetes is a complex multilevel process resulting in a set of molecular-genetic, physiological-biochemical, and morphological-ultrastructural changes in the cells. When the xylotrophic basidiomycetes Lentinus edodes, Grifola frondosa, and Ganoderma lucidum were grown on wood waste as the substrate, the ultrastructural morphology of the mycelial hyphal cell walls differed considerably between mycelium and morphostructures. As the macromycetes passed from vegetative to generative development, the expression of the tyr1, tyr2, chi1, chi2, exg1, exg2, and exg3 genes was activated. These genes encode enzymes such as tyrosinase, chitinase, and glucanase, which play essential roles in cell wall growth and morphogenesis