Pronounced Stimulatory Effect Research Articles

Enhanced fig tree production through endomycorrhizal fungi inoculation: A nursery study on tree cuttings derived from the Ifrane Region, Morocco

The mycorrhizal association represents a crucial symbiotic bond necessary for the vitality of the majority of plants. This report aims to investigate the impact of endomycorrhizal fungi on the production of fig trees from cuttings. The research took place in a nursery environment by inoculating fig cuttings sourced from three sites in Morocco with a composite inoculum of arbuscular mycorrhizal fungi (AMF). The mean root and vegetative masses of the inoculated plants after 140 days were 33.1 g and 35 g compared with 1.8 g and 6.4 g in the control. The mean shoot length and number of leaves developed from treated cutting were 98.5 cm and 17, respectively 29 cm and 4 leaves in control. Thus, the newly formed roots showed a well-established mycorrhizal colonization and contained different structures characteristic of arbuscular endomycorrhizae: arbuscules, vesicles, endophytes, hyphae and spores. The frequency and intensity of root mycorrhization were approximately 80% and 15.5%, respectively. The contents of arbuscules and vesicles were 2.5% and 5.5%. The study of formed spores showed a combination of 52 spores per 100 g of soil, belonging to 11 species and 3 genera: Glomus, Acaulospora and Rhizophagus with dominance of the genus Glomus (85%). Glomus macrocarpum and Glomus deserticola were the most abundant species, appearing at 30.77% and 19.23%, respectively. The results demonstrate a pronounced stimulating effect on both root formation and vegetative shoot development in the inoculated cuttings. Specifically, the early inoculation of fig cutting with the AMF inoculum significantly enhanced rhizogenesis end root system development. Consequently, the gradual establishment of mycorrhizal symbiosis at the root level of inoculated plants led to good development of the root mass and the vegetative mass. The inoculated plants showed good root and vegetative mass development due to the installation and the progressive development of mycorrhizal symbiosis at their roots.

Histone H3K9 Methylation Features under Hypoxic Conditions after the HIF1A Knockdown in Mesenchymal Stromal Cells In Vitro.

The effects of HIF1A knockdown by RNA interference on the histone H3K9 methylation in human umbilical cord mesenchymal stromal cells in vitro under conditions of 24-h exposure to hypoxia (1% O2) were studied. Evaluation of transcriptional activity of genes involved in the regulation of H3K9 methylation (KDM3A, KDM4A, and EHMT2) and the cytofluorimetric analysis of the expression of the corresponding antigens and H3K9 methylation level demonstrated a pronounced stimulating effect of hypoxic exposure. Moreover, the expression of KDM4A and EHMT2 was regulated by HIF1A-mediated mechanism, unlike KDM3A; the level of the corresponding proteins depended on HIF1A. In addition, the HIF-1-dependent regulation of KDM3A, KDM4A, and EHMT2/G9a, and directly the H3K9 methylation level in mesenchymal stromal cells also took place under normoxia conditions.

Morphological characteristics of the stromal-parenchymatous component of the liver of fetal and newborn developed under the conditions of maternal hypertension disease

Background. Extragenital pathology of the mother can cause various changes in the body of the newborn, which in the future are the cause of violations in physical and mental development, and also causes an increase in somatic and infectious diseases in such children. Therefore, timely diagnosis of liver diseases in newborns and young children remains an urgent problem today. Purpose – the article is devoted to the study of the morphological features of the stromal-parenchymal component of the liver of fetuses and newborns that developed under conditions of maternal hypertension. Materials and Methods. Research was conducted on liver preparations of fetuses and newborns who died at the age of 37 weeks of gestation – the 1st day of postnatal life. All the studied material was divided into two groups. The control group (group K) included fetuses and newborns from healthy mothers (18 cases) who died due to acute disruption of umbilical-placental circulation and birth trauma. The comparison group (group C) consisted of 36 fetuses and newborns from mothers with stage II hypertension. The obtained micropreparations were stained with hematoxylin and eosin, picrofuchsin according to the van Gizon method, and according to the Mallory method. The parenchymal, stromal, and vascular components were calculated as a percentage, and then the stromal-parenchymal ratio was calculated. Results. The results we obtained during the conducted research indicated that the influence of chronic intrauterine hypoxia, which developed against the background of stage II hypertensive disease in the mother, leads to the development of destructive-dystrophic changes in the liver tissue of fetuses and newborns. Chronic intrauterine hypoxia with maternal arterial hypertension leads to parenchymal losses and has a pronounced stimulating effect on the stromal component of the liver, and this can lead to a significant decrease in the functional activity of the liver under the influence of certain triggers and increases the risk of developing liver failure and liver cirrhosis in the future. Conclusions. During our research, it was established that chronic intrauterine hypoxia, which develops against the background of stage II hypertensive disease in the mother, leads to the development of significant destructive-dystrophic changes in the liver tissue of fetuses and newborns. There is a decrease in the volume of the parenchyma due to the volume of hepatocytes and an increase in the volume of the stroma with vessels due to the volume of the portal tracts (11.2 ± 2.2% compared to the control – 3.4 ± 0.3%), the volume of the central veins (12.9 ± 1.3% compared to the control – 9.4 ± 0.4%), the volume of sinusoids (13.3 ± 1.1% compared to the control – 11.3 ± 0.8%). It was established that the morphological changes described by us in the liver of fetuses and newborns in the subsequent ontogenesis may become a prerequisite for the development of hepatobiliary system pathology in such children.

Influence of milk phospholipids on microstructural changes in rat liver under tetracycline-induced hepatosis

The growing number of cases of adverse effects of medicines on the liver leads to hepatopathology and the development of complications such as cirrhosis, necrosis, liver failure, and liver carcinoma. Therefore, the purpose of this study was to determine the characteristic structural changes in the liver of rats with tetracycline damage and to find out the corrective effectiveness of milk phospholipids. The study performed histological examination of sections from different parts of the liver in experimental rats, which were stained with haematoxylin and eosin according to the conventional method. It was found that in case of artificial modelling of tetracycline-induced hepatosis in rats, the general architecture of the liver is preserved. At the same time, large-scale damage to hepatocytes and the development of fatty and granular dystrophy were recorded in the affected animals. Some of the damaged cells were destroyed, followed by lysis of fragments of the destroyed cells. The described microscopic changes were most pronounced in the areas of the liver under its capsule. As a result, there was partial or complete disorganisation of the liver lobes in all lobes of the organ. The use of milk phospholipids in the form of a biologically active additive “FLP-MD” as a corrective therapy in sick rats prevented the development of hepatocyte dyscomplexity, contributed to a substantial reduction in the count of destroyed cells in a state of dystrophy with isolated cases of disordered organisation of the liver laminae. This suggests a pronounced stimulatory effect of milk phospholipids on the processes of hepatocyte regeneration and repair in tetracycline-induced liver damage in rats. In case of separate administration of a milk phospholipid-based supplement to clinically healthy animals, the microscopic structure of liver samples did not differ from that of the control group. The findings of this study are of practical value for pathologists, clinicians, and therapists in case of development of drug-induced hepatopathology in animals, especially when using tetracycline antibiotics, and for determining the strategy in the treatment of such patients

The effects of sub-central development on the economic growth of provincial capitals: A study based on the DID and PSO-RF models

As a core component in deepening reforms and expanding the strategy of dual circulation development, the importance of developing sub-centers for the economic growth of provincial capitals is self-evident. This paper, based on panel data from 300 prefecture-level cities in China from 1998 to 2020, constructs a progressive double-difference (DID) model to assess the impact of sub-central development on the economic growth of provincial capitals. The research reveals that the development of sub-centers at the provincial level significantly promotes the economic growth of provincial capitals. Moreover, the policies of sub-center cities have a more pronounced stimulating effect on the economic growth of provincial capitals in the central and western regions. Additionally, the more sub-center cities there are, the more apparent the effect on promoting the economic growth of provincial capitals. Subsequently, this paper employs the Particle Swarm Optimization - Random Forest (PSO-RF) model through machine learning to validate the above conclusions, confirming the validity of the findings.

Нанопрайминг семян сои с применением марганецсодержащего нанокомпозита в арабиногалактановой матрице

The results of studies on the stimulation of soybean seed germination under the influence of nanopriming with a chemically synthesized nanocomposite (NC) based on manganese-containing nanoparticles and a water-soluble polysaccharide arabinogalactan are presented. The study was carried out on the phytopathosystem: soybean seeds Glycine max (L) – phytopathogenic bacterium Pectobacterium carotovorum. Phytopathogenic bacterium P. carotovorum causes the infectious disease soft rot in various plant species from vegetables to trees and shrubs. After nanopriming (treatment Mn(OH)2/AGs NC), seed germination, biometric characteristics (mass and length of the root and hypocotyl) and biochemical parameters of soybean seedlings (content of reactive oxygen species (ROS), activity of antioxidant enzymes, amount of lipid peroxidation products) were analyzed. It was shown that the infection of seeds with P. carotovorum led to a decrease in their germination, 50 % of the seeds did not germinate. In addition, seedlings obtained from infected seeds had reduced all biometric characteristics compared to the control. Infection of seeds with P. carotovorum showed an increase in the amount of ROS in root tissues and caused a significant increase in peroxidase activity in the roots of soybean seedlings. A significant increase in diene conjugates (DC) was noted in the tissues of the roots of the hypocotel in this variant. The nanocomposite of manganese hydroxide and sulfated arabinogalactan (Mn(OH)2/AGs NC, 4,8 % Mn) removed the negative effect of the phytopathogen P. carotovorum. Morphometric characteristics of seed sprouts after nanopriming were characterized by an increase in biomass compared to the control. Thus, after nanopriming with Mn(OH)2/AGs NC, the length and weight of the root in uninfected seeds increased. The most pronounced stimulating effect of nanopriming was expressed in sprouts obtained from infected seeds. The length weight of the hypocotyl and the length of the root in infected seeds significantly increased after nanopriming of infected seeds. The precursor NC – MnSO4×5H2O did not affect the germination and sprouting of seeds both infected with P. carotovorum and free from infection. Nanopriming of infected seeds reduced the formation of ROS in seedling tissues. It was found that after nanopriming, peroxidase activity increased significantly in soybean roots compared to the control. This effect can be explained by the antioxidant effect of Mn NPs. Nanopriming of infected seeds reduced the amount of DC in root tissues to the control level. Thus, the mechanisms of interaction of NC with the studied phytopathosystem are represented by an increase in seed germination under the influence of NC, stimulation of morphometric parameters of seedlings, as well as an effect on the components of the antioxidant system. The obtained results and our previous studies on the antibacterial effect of Mn(OH)2/AGs against phytopathogens Clavibacter sepedonicus demonstrate the great potential of using Mn(OH)2/AGs in practical agriculture, in particular for application in the field of stimulating plant growth and increasing their resistance to phytopathogens.

Нанопрайминг семян сои с применением марганецсодержащего нанокомпозита в арабиногалактановой матрице

The results of studies on the stimulation of soybean seed germination under the influence of nanopriming with a chemically synthesized nanocomposite (NC) based on manganese-containing nanoparticles and a water-soluble polysaccharide arabinogalactan are presented. The study was carried out on the phytopathosystem: soybean seeds Glycine max (L) – phytopathogenic bacterium Pectobacterium carotovorum. Phytopathogenic bacterium P. carotovorum causes the infectious disease soft rot in various plant species from vegetables to trees and shrubs. After nanopriming (treatment Mn(OH)2/AGs NC), seed germination, biometric characteristics (mass and length of the root and hypocotyl) and biochemical parameters of soybean seedlings (content of reactive oxygen species (ROS), activity of antioxidant enzymes, amount of lipid peroxidation products) were analyzed. It was shown that the infection of seeds with P. carotovorum led to a decrease in their germination, 50 % of the seeds did not germinate. In addition, seedlings obtained from infected seeds had reduced all biometric characteristics compared to the control. Infection of seeds with P. carotovorum showed an increase in the amount of ROS in root tissues and caused a significant increase in peroxidase activity in the roots of soybean seedlings. A significant increase in diene conjugates (DC) was noted in the tissues of the roots of the hypocotel in this variant. The nanocomposite of manganese hydroxide and sulfated arabinogalactan (Mn(OH)2/AGs NC, 4,8 % Mn) removed the negative effect of the phytopathogen P. carotovorum. Morphometric characteristics of seed sprouts after nanopriming were characterized by an increase in biomass compared to the control. Thus, after nanopriming with Mn(OH)2/AGs NC, the length and weight of the root in uninfected seeds increased. The most pronounced stimulating effect of nanopriming was expressed in sprouts obtained from infected seeds. The length weight of the hypocotyl and the length of the root in infected seeds significantly increased after nanopriming of infected seeds. The precursor NC – MnSO4×5H2O did not affect the germination and sprouting of seeds both infected with P. carotovorum and free from infection. Nanopriming of infected seeds reduced the formation of ROS in seedling tissues. It was found that after nanopriming, peroxidase activity increased significantly in soybean roots compared to the control. This effect can be explained by the antioxidant effect of Mn NPs. Nanopriming of infected seeds reduced the amount of DC in root tissues to the control level. Thus, the mechanisms of interaction of NC with the studied phytopathosystem are represented by an increase in seed germination under the influence of NC, stimulation of morphometric parameters of seedlings, as well as an effect on the components of the antioxidant system. The obtained results and our previous studies on the antibacterial effect of Mn(OH)2/AGs against phytopathogens Clavibacter sepedonicus demonstrate the great potential of using Mn(OH)2/AGs in practical agriculture, in particular for application in the field of stimulating plant growth and increasing their resistance to phytopathogens.

Effect of Paracetamol Biodegradation Products Quantitative Indicators of the Anatomical Structure of the Lily of the Valley Leaf (<i>Convallaria majalis</i> L.)

Introduction. Due to the global pharmaceutical pollution of water bodies in many countries, there is currently an increasing interest of researchers in finding effective, including microbiological, ways of processing pharmaceutical waste to obtain new useful products based on them. The results of previous studies have shown that the product of bacterial destruction of paracetamol exhibits pronounced stimulating properties in relation to medicinal plants of the aster, linseed, clear-flowered, plantain, etc. families. There is no information about the effect of this product on plants of the liliaceae family.Aim. To investigate the effect of the paracetamol biodegradation product on the quantitative indicators of the anatomical structure of the lily of the valley leaf – Convallaria majalis L., fam. Liliaceae.Materials and methods. The experiments were laid in the Kirov region (2021, 2022) in accordance with the Guidelines for Conducting Registration Tests of Growth Regulators (2016). The scheme of the experiment: the control area – natural watering with water; the test area – watering with the product of biodegradation of paracetamol obtained on the basis of the laboratory of alkanotrophic microorganisms of the PFRC UB RAS (Perm); the reference area – watering with the growth stimulator "Zircon" (NRPP "NEST M", Russia). Quantitative parameters of the anatomical structure of the lily of the valley leaf were evaluated by the linear dimensions of the epidermis, elements of the conducting system and mesophyll. Viewing, photographing and processing of photographs was carried out using a microscope Motic (Motic Deutschland GmbH, Germany) in the program Motic Image 2000. Mathematical processing of experimental data was carried out by the method of variance analysis using Excel 2019, STATISTICA 12.6, STATGRAPHICS Plus 5.1 programs.Results and discussion. Anatomical examination showed an increase in the size of the cells of the upper and lower epidermis of the lily of the valley leaf in the designated growing seasons on the accounting sites treated with a product of the biodegradation of paracetamol and a standard ("Zircon") in comparison with the control (water). When using the biodegradation product of paracetamol, an increase in the thickness of the mesophyll by 10–16 % and the number of layers by 14–30 % was recorded. When using the growth stimulator "Zircon", the thickness of the mesophyll and the number of its layers increased by 3–6 % and 8–15 %, respectively. The increase in the thickness of the phloem and xylem of the conductive beam when treated with the paracetamol biodegradation product was 20–30 % greater than in the control, and 2 times greater than when treated with the growth stimulator "Zircon".Conclusion. The product of the biodegradation of paracetamol in different years of use, regardless of environmental factors, shows a pronounced stimulating effect on the lily of the valley of the May, significantly increasing the quantitative indicators of the anatomical structure of the leaf compared with the growth stimulator "Zircon" and control (water). The obtained data can be used in medicinal plant growing in the development of agrotechnical methods for growing raw materials of the lily of the valley of the May, as well as in the formation of a dossier when registering this product as a new plant growth stimulator.

RESEARCH ON VARIOUS MODELS OF THE BIOLOGICAL EFFECTS OF COMPONENTS OF A COMPLEX EFFECT (PHOTOIRRADIATION; EXOSOMES OF MESENCHYMAL STEM CELLS AND NANOPARTICLES) FOR THE CORRECTION OF THE INFLAMMATORY PROCESS

RESEARCH ON VARIOUS MODELS OF THE BIOLOGICAL EFFECTS OF COMPONENTS OF A COMPLEX EFFECT (PHOTOIRRADIATION; EXOSOMES OF MESENCHYMAL STEM CELLS AND NANOPARTICLES) FOR THE CORRECTION OF THE INFLAMMATORY PROCESS

Novel Stimulants of Medicinal Basidiomycetes Growth Based on Nanoparticles of N-monosubstituted Amino Acid Derivatives of Fullerene C60

The influence of nanoparticles of hydrated C60 fullerene and its N-monoamino acid derivatives on the oxidative metabolism and growth of the mycelial biomass of basidiomycetes during their submerged cultivation was studied. It was found that the supplementation of culture media with nanoparticles of the studied compounds at their final concentration range of 10−7 to 10−11 M significantly increased the resulting biomass, while the severity of the effect in this concentration range changed slightly. That prompted the use of nanomolar concentrations of compounds as reasonable. The most pronounced stimulating effect (an increase in biomass of about 240% with respect to control) was observed when culturing Laetiporus sulphureus, the intrinsically high level of oxidative metabolism of which was significantly lowered by the presence of the studied additives. It was shown that the growth-enhancing action of nanoparticles of fullerene C60 and its derivatives could not be attributed to photochemical reactions, particularly fullerene photoexcitation. Fullerene and its derivatives manifest a growth regulatory effect on bio-objects from different kingdoms of the living world (plants and fungi), which is indicative of these compounds’ mechanism of action based on a direct impact on fundamental, universal for all living beings, biophysical processes, primarily chain free-radical oxidation.

Ultrasound-assisted green syntheses of novel pyrimidine derivatives and their comparison with conventional methods

Ultrasound-assisted green syntheses of novel potentially bioactive pyrimidine derivatives have been carried out. The same compounds were obtained by conventional methods of synthesis, and the reaction times and yields of final products obtained by these two methods were compared. It was found that the time of utrasound-promoted reactions was reduced by almost 6–96 times, and their yields were equal or turn out to be greater compared to the traditional approach. The synthesized compounds showed a pronounced stimulating effect on plant growth. The most active derivatives were selected for deeper biological studies and subsequent field trials.

Aminodihydrophthalazinedione Sodium Transdermal Therapeutic System Specific Activity on an ExperimentalModel of Extensive Liver Resection

Previously, the authors showed that the application of the aminodihydrophthalazinedione sodium (ADPS) immunomodulator transdermal therapeutic system (TTS) to laboratory animals provides bioavailability analogous to the intramuscular administration of this drug at the same dose. At the same time, its maximum blood concentration is significantly reduced, and the retention time of the drug in the body is increased more than 10-fold, which can contribute to prolonging the drug effect. The aim of the work was to identify a possible positive effect of the transdermal administration of the ADPS immunomodulator on reparative liver regeneration on an experimental model of extensive liver resection (ELR). It has been shown that at a period of 48 h after ELR, the percutaneous administration of the immunomodulator has a pronounced stimulating effect on the mitotic activity of rat liver cells; by 72 h after ELR, an accelerated rate of recovery of hepatic homeostasis in the body was observed in laboratory animals in groups with the application of the ADPS TTS versus the control group.

Role of glutamate in thymic energy metabolism

Objective. To analyze the energetic role of glutamate in thymocytes and tissues of the thymus at different stages of its age-related involution.Materials and methods. The studies were performed on white male rats. The state of energy metabolism was determined by the rate of oxygen consumption by thymus tissues and thymocytes using a polarographic method.Results. It was found that in the thymus tissues of 4, 5 and 6-month-old rats the rate of oxygen consumption is constant and does not significantly change in response to glutamate administration. Thymocytes - the immunocompetent thymus cells of 3- and 8-month-old animals, despite similar levels of oxygen consumption on endogenous substrates, respond differently to glutamate action. In thymocytes of 3-month-old animals, the action of glutamate has a more pronounced stimulating effect on bioenergetic processes compared to 8-month-old animals. There is a reason to believe that the decrease in the efficiency of glutamate action as the animals grow older is caused by the processes of age-related involution of the thymusConclusion. Glutamate stimulates aerobic respiration in the thymocytes of 3- and 8-month-old animals, with a greater stimulating effect in the thymocytes of younger animals. At the same time, glutamate does not cause significant changes in the rate of oxygen consumption in the thymus tissues of 4-, 5-, and 6-month-old animals.

ВЛИЯНИЕ ФИТОПРЕПАРАТА НА ОСНОВЕ ХВОИ НА ИММУНИТЕТ ЦЫПЛЯТ-БРОЙЛЕРОВ ПРИ ВАКЦИНАЛЬНОМ СТРЕССЕ

The article presents the results of studying the effect of a phytopreparation based on needles on the cellular and humoral immunity of broiler chickens of the ROSS 308 cross under vaccine stress. For the experiment, 4 groups of 25 heads were formed. Chickens of the control group were on the main diet. In the diet of the 1st experimental group, a phytopreparation based on needles was introduced with water, the 2nd and 3rd - with food. At the age of 14 days, the chickens were vaccinated against Newcastle disease (ND). In broiler chickens vaccinated against ND and receiving the phytopreparation with feed, a more pronounced stimulating effect was noted, characterized by an increase in the number of T-lymphocytes by 1.3-1.8 times, cytotoxic T-lymphocytes - by 1.1-1.9 times , as well as the activity of cationic proteins - by 0.1-0.6 c.u. When drinking a complex of biologically active substances, only activation of intracellular antimicrobial peptides was observed, but in a longer period after vaccination.

Phyto-stimulating Effect of Paracetamol Biodestruction Product on Calendula Officinalis

Introduction. In recent years, there has been an increase in fundamental interest in the search for effective, including microbiological, methods for processing pharmaceutical waste to obtain new biologically active compounds on their basis. Our results showed that the product of bacterial degradation of paracetamol (BDP) exhibits pronounced stimulating properties in relation to medicinal plants of the families Plantaginaceae, Lamiaceae, Urticaceae, Linaceae and can be used as an inducer of the accumulation of biologically active substances in them.Aim. The purpose of this work is to study the effect of BDP on the dynamics of biomass accumulation and the content of flavonoids in the flowers of Calendula officinalis L., fam. Asteraceae in comparison with the control (water) and growth stimulator "Zircon".Materials and methods. BDP obtained on the basis of the Laboratory of Alkanotrophic Microorganisms Institute of Ecology and Genetics of Microorganism UB RAS (Perm). Calendula officinalis flowers collected from plants treated with BDP and Zircon growth stimulator were used to study the dynamics of dry biomass accumulation and changes in the content of flavonoids by the spectrophotometric method. For a comparative analysis of the intensity of the increase in the biomass of flowers during the treatment of plants with these agents, as well as to predict the accumulation of flavonoids, kinetic modeling was used.Results and discussion. The total biomass harvest of calendula officinalis flowers when treated with BDP increased by 55 %, and when treated with the Zircon growth stimulator, by 24 % compared to the control. The content of flavonoids in flowers when treated with these agents increased by 101 and 40 %, respectively. The dates for the beginning of the collection of calendula officinalis flowers in the conditions of the Western Urals were determined: with the use of the Zircon growth stimulator – from July 20, BDP – from August 1, without treatment with growth stimulants – from August 14. Both stimulants increase the end date of the collection of raw materials by 10 days.Conclusion. The product of bacterial degradation of paracetamol has a pronounced stimulating effect on calendula officinalis, significantly increasing the biomass of flowers and the content of flavonoids in them compared to the control (water) and growth stimulator "Zircon". With the use of kinetic modeling, the values of the parameters of the growth rate of the biomass of plant raw materials during the treatment of plants with these agents were established, a forecast was made for the accumulation of flavonoids, and the timing of the collection of calendula officinalis flowers in the conditions of the Western Urals was substantiated.

EFFICIENCY OF OILY AND POLYELECTROLYTE ADJUVANTS IN CASE OF RABBIT HYPERIMMUNIZATION WITH DIMETHYLSULFOXIDE ANTIGEN OF PSEUDOTUBERCULOSIS MICROORGANISM

A comparative evaluation of two adjuvants: polyazolidinammonium modified with iodine hydrate ions (PAAH) and complete Freud's adjuvant (CFA) was conducted. The assessment was carried out in the process of hyperimmunization of rabbits with a mixture of an adjuvant and dimethyl sulfoxide antigen (DA) of a pseudotuberculous microbe. Both adjuvants stimulated antibody genesis and allowed to obtain hyperimmune sera with high antibody titers. However, CFA showed a slightly higher activity (1:819200) than PAAH (1:409600). The most pronounced stimulating effect of adjuvants was expressed at low doses of DA (0.2-1 mg/rabbit). Large doses of the antigen (16.0 mg/rabbit) enabled to obtain sera with a high antibody titer without adjuvants. The stimulating effect of DA-PAAH and DA-CFA complexes on antibody genesis is associated with the rate and quality of lymphocyte differentiation, and to a lesser extent with an increase of their number. PAAH showed a weak irritating effect on body tissues at the injection spot, and CFA had a strong irritating effect. The application of CFA led to formation of many painful indurations in the subcutaneous tissue of rabbits, which did not resolve between immunizations. Thus, usage of PAAH as an adjuvant in hyperimmunization of rabbits with DA of the pseudotuberculous microbe is potentially productive and without CFA disadvantages.

M-CSF and GM-CSF determinate fibromodulatory activity of polarized human macrophages

GM-CSF and M-CSF, the hematopoietic colony-stimulating factors, induce various phenotypic changes in macrophage lineage populations and promote cell differentiation, respectively, into M1- and M2-like macrophages. The pro- and anti-inflammatory properties of macrophages generated by these colony-stimulating factors are well described, but the contribution of differentiation and polarization signals to the fibromodulatory activity of macrophages remains unexplored. To clarify the differences in the fibrogenesis regulation mechanisms inherent in differently activated macrophages, we studied the effects of macrophage-conditioned media on proliferation and differentiation of dermal fibroblasts. In this study, the human macrophages generated from peripheral blood monocytes were investigated. They were induced for differentiation by M-CSF or GM-CSF, being further polarized in the M1 direction with lipopolysaccharide and, in the M2 direction, with IL-4 or dexamethasone. Proliferative response of the fibroblasts was determined radiometrically by [3H]-thymidine incorporation. Differentiation into myofibroblasts was determined with flow cytometry technique, as expression of a specific marker α-smooth muscle actin (α-SMA). The level of macrophage TGF-β1 production was assessed using an appropriate ELISA kit. The data obtained indicate that the macrophages differentiated under the influence of “homeostatic” M-CSF are characterized by a moderate stimulating effect upon fibroblast proliferation, and the effects of M2 (IL-4) and M2 (Dex) macrophages exceed that of M1 (LPS), but do not differ significantly from each other. The M-CSF-induced M1 (LPS) and M2 (IL-4) macrophages, but not M2 (Dex), enhance the fibroblast differentiation and show similar level of stimulation. In contrast to M-CSF, the macrophages induced by “pro-inflammatory” GM-CSF exhibit a pronounced stimulatory effect on fibroblast proliferation, and the effects of M2 macrophages exceed those of M1 cells, being most pronounced for M2 (Dex). At the same time, only GM-CSF-induced M2 (IL-4) macrophages enhance fibroblast differentiation. Dexamethasone-polarized macrophages do not significantly affect fibroblast differentiation regardless of the CSF used (M-CSF or GM-CSF). The content of TGF-β1 in the supernatants of differently activated macrophages does not correlate with the level of stimulating effect of macrophage-conditioned media upon fibroblast differentiation. In general, the data obtained suggest the involvement of differentiation and polarization signals into modulation of pro- and anti-fibrogenic properties of macrophages.

The effect of biologically active substances on the intestinal microflora of quails

The Intestinal's microflora has a pronounced stimulating and immunomodulatory effect on the body. The use of an experimental group of quails of the Estonian breed Chiktonik at a dose of 1 ml / l of water from the 1st to the 10th day and from the 31st to the 40th day. Vetom 1.1 at a dose of 25 mg/kg of body weight from the 11th to the 20th and from the 41st to the 50th day showed a change in the coprogram, the qualitative and quantitative composition of the microflora compared with the control. The determination of the species composition of the microflora was carried out by inoculation of dilutions of the intestinal contents on differential diagnostic and selective nutrient media, the study of feces was carried out with a set of reagents "Clinic-feces". In 30- and 60-day-old quails of the experimental group, the pH of the content is acidic, there is no intracellular starch, digestible fiber, neutral fat, single cocci and rod-shaped microflora are noted; by the age of 60 days, single leukocytes and erythrocytes appear. In the control group, the pH of the content is alkaline, there is extra- and intracellular starch, digestible and indigestible fiber, drops of neutral fat, coccal and rod-shaped microflora, iodophilic microflora, cellular elements, mucus. In 30-day-old quails of the experimental group, an increase in the number of Lactobacillus spp. and Bifidobacterium spp. up to 6.6ґ106 and 1.0?105 Cfu/g compared to 2.6ґ106 Cfu/g and 2.8ґ104 in the control group; the number of Escherichia coli and Enterococcus faecalis decreased and amounted to 1.08?106 CFU/g and 1.0ґ104 CFU/g, respectively, against 3.15ґ106 CFU/g and 7.0?104 CFU/g in the control group. In 60-day-old quails of the experimental group Escherichia coli, Enterococcus faecalis, Bacillus spp., Lactobacillus spp. and Bifidobacterium spp. were 5.8ґ104, 1.87ґ105, 1.0?103, 7.6ґ106 and 1.49ґ105 CFU/g versus 1.6ґ105, 2.86ґ105, 2.0ґ103, 2.88ґ106 and 3.2ґ104 CFU/g in the control group. Therefore, the developed scheme for the use of biologically active substances contributes to the reduction of opportunistic microflora and successful colonization of the small intestine by resident microflora.

Serotonin syndrome onset following 25H-NBOH and MDPHP consumption: A case report

The continual emergence of new psychoactive substances remains a global health issue. We reported a case of serotoninergic syndrome following the consumption of 3,4-methylenedioxy-α-pyrrolidinohexanophenone (MDPHP) and 25H-NBOH documented by liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS). A 23-year-old man, with a history of cathinone regular use, presented a seizure and a loss of consciousness 30 min after the consumption of a powder identified as 25I-NBOH. The way of exposure was not documented. On arrival at the mobile emergency unit, the patient exhibited a serotonin syndrome with hyperthermia at 41 °C, hypertonia of the upper limbs, bilateral mydriasis, Glasgow coma 3, hypertension at 16/8, tachycardia at 180 bpm, and SaO 2 = 50%. During the hospitalization, he developped acute renal failure, acute liver failure, rhabdomyolysis and disseminated intravascular coagulation. He was discharged after 20 days of hospitalization but kept neurological sequelae. Following his hospital admission, blood and urine samples were drawn and sent to the toxicology laboratory to determine the cause of poisoning. In total, 200 μL of plasma and urine samples were extracted with QuEChERS salts and acetonitrile. The extracts were analyzed by liquid chromatography hyphenated to high-resolution mass spectrometry (Orbitrap, Exploris 120) allowing both a non-targeted screening and the simultaneous quantification of 132 compounds frequently found in poisonings. Analysis was realized in full scan experiment with a nominal resolving power of 60,000 FWHM within the 125–650 m/z mass range, followed by four cycles of DDA with a mass resolution of 16,000 FWHM. Pharmaceuticals administered to the patient during his medical care were found in plasma and urine: midazolam, hydroxyzine and its metabolite cetirizine, ketamine, etomidate, laudanosine (atracurium metabolite), lidocaine. Additionally, two NPS were identified and then confirmed with certified reference materials (CRM): MDPHP in plasma and urine, and 25H-NBOH in urine. 25H-NBOH was not detected in plasma despite a limit of detection being 0.01 ng/mL. Unfortunately, there were not enough remaining samples after the screening analyzes, therefore the quantification could not be carried out. In order to estimate the concentration, a calibration curve was carried out with the two compounds under the same conditions as the screening analysis (same pretreatment, same internal standards at the same concentrations, as well as the same analytical conditions), and the data corresponding to the initial acquisition was then reprocessed. MDPHP concentration was 1 ng/mL in plasma and 48 ng/mL in urine, 25H-NBOH concentration was 0.4 ng/mL in urine. 25I-NBOH was specifically searched in both samples and was not detected. MDPHP is a synthetic cathinone of the pyrovalerone type (e.g. MDPV), it acts as a stimulant on the release of dopamine and norephedrine and simultaneously as reuptake inhibitor, causing a pronounced stimulatory effect on the central nervous system (Grapp M, et al., Drug Test Anal 2020;12:1320–35). 25H-NBOH is a psychedelic belonging to the NBOMe family, they are potent agonists of 5-HT2A receptor (Pottie et al., Biochem Pharmacol 2020;182:114251). The clinical manifestations observed here are similar to those described in cases of NBOMe poisoning, and are probably attributable to 25H-NBOH, they may have been potentiated by the presence of MDPHP (Kyriakou C, et al., Eur Rev Med Pharmacol Sci 2015;19:3270–81). To our knowledge, very few study have been dedicated to MDPHP poisoning, and this is the first describing 25H-NBOH poisoning.

Prebiotic effect of galacto-oligosaccharides on the skin microbiota and determination of their diffusion properties.

Recently, prebiotics are attracting plenty of attention in the field of skin care, since it is found that they are able to support the balance of beneficial and harmful microorganisms on the skin, and accordingly prevent several skin conditions associated with microbial imbalance. Topical application of prebiotics, although insufficiently investigated, holds great promise in improving skin health. The purpose of this research was to determine the prebiotic potential of galacto-oligosaccharides (GOS) for skin microbiota and suitability for incorporation in different topical formulations, and finally, provide insights into their diffusion properties. The prebiotic effect of GOS was evaluated through the influence on the growth of Staphylococcus epidermidis and Staphylococcus aureus, the most common resident and pathogenic bacterium of the skin microbiota, respectively. Also, with the future use of GOS in cosmetic products in mind, the diffusion of GOS molecules from two different topical formulations, hydrogel and oil-in-water (O/W) gel emulsion, was monitored employing Franz diffusion cell and two systems-with cellulose acetate membrane and transdermal diffusion test model, Strat-M® membrane. Course of fermentation and the amount of diffused GOS molecules were monitored using high-performance liquid chromatography (HPLC). The in vitro results revealed that GOS at a concentration of 5% (w/v) has a pronounced stimulatory effect on S.epidermidis, while simultaneously showing an inhibitory effect on S.aureus, both in nutrient broth and cosmetic formulations. GOS trisaccharide and tetrasaccharide diffusion coefficients from O/W gel emulsion were calculated to be 5.61·10-6 cm2 s-1 and 1.41·10-8 cm2 s-1 , respectively. The diffusion coefficient of GOS trisaccharides from hydrogel was 3.22·10-6 cm2 s-1 , while it was not determined for tetrasaccharides due to low diffused concentration. Transdermal diffusion tests revealed that GOS incorporated in two formulations stays at the surface of the skin even after 24h. When applied in adequate concentration, GOS has the potential to be used as a skin prebiotic. Novel GOS enriched formulations, Aristoflex® AVC-based hydrogel and Heliogel™-based O/W gel emulsion, provided efficient diffusion and delivery of prebiotic GOS molecules to the skin surface.