Cytokines and their regulators in rat lung following scorpion envenomation.

Nowadays, more than two billion inhabitants of underdeveloped tropical and subtropical countries are at risk of being stung by scorpions. Scorpion stings annually cause 2000-3000 deaths as they can lead to the respiratory and/or cardiovascular complications. Pathogenesis of lung damage under scorpion envenomation is often comprehensive. Respiratory failure can have a cardiogenic origin, associated with venom neurotoxin action. However, some venom components can stimulate pro-inflammatory signaling cascades followed by cytokines synthesis, recruit and activate immune cells, participating in the inflammatory response in lung injury. Scorpions of the Leiurus genus ("deathstalker") are one of the most dangerous Arthropoda. To date, 22 species of this genus have been described, but the venom composition and the mechanisms of tissues damage under envenomation have been studied to some extent only for L. quinquestriatus, L. hebraeus, and L. abdullahbayrami. Scorpions of L. macroctenus species are expected to be very hazardous, but the possibility of their venom cause inflammation in the lung tissue has not been investigated to date. Therefore, in this study, we focused on evaluating the levels of cytokines and their regulators - transcription factors (HIF-1α and NF-κB) and growth factors (FGF-2, VEGF, and EGF) - in rat lung homogenates after L. macroctenus envenomation. The results revealed a decrease in the levels of most pro-inflammatory cytokines (IL-6, IL-8, IL-1ß and TNF-α) with simultaneous rise in the content of both anti-inflammatory cytokines (IL-4 and IL-10) and interferon-γ. Furthermore, the levels of all researched transcription factors and growth factors were shown to be increased too. The detected changes peak occurred at 24 h, whereas a tendency towards all indicators values normalization was observed in 72 h after venom injection. Thus, our results did not reveal signs of a classic inflammatory process in the lungs of rats injected with L. macroctenus venom. However, the obtained data indicate venom influence both on cytokine profile and on their regulators content in the rat lungs, which is a feature of certain alterations in the innate immune response, caused by studied venom components. But, the mechanisms of the changes we found require additional researches.

Features of immune reactivity of the spleen and mechanisms of organ damage under the influence of animal venom toxins including scorpions (review).

OBJECTIVE: Aim: To establish features of immune reactivity of the spleen and mechanisms of organ damage under the influence of animal venom toxins including scorpions. PATIENTS AND METHODS: Materials and Methods: A thorough literature analysis was conducted on the basis of PubMed, Google Scholar, Web of Science, and Scopus databases. When processing the search results, we chose the newest publications up to 5 years old or the most thorough publications that vividly described the essence of our topic. CONCLUSION: Conclusions: Spleen plays a leading role in the implementation of the body's defense processes, the elimination of structural elements affected by toxins, and the restoration of immune homeostasis. Its participation in the formation of the immune response can be accompanied by qualitative and quantitative changes in histological organization. Morpho-functional changes in the spleen under the action of animal venom toxins currently require careful study, because from the information available in the literature today, it is not possible to clearly construct a complete picture of lesions of certain components of the organ at the microscopic or submicroscopic levels. Therefore, this direction of research in the medical field is currently relevant, taking into account the existence of a large number of poisonous animals, including scorpions.

Effect of animal venom toxins on the main links of the homeostasis of mammals (Review).

The human body is affected by environmental factors. The dynamic balance between the organism and its environment results from the influence of natural, anthropogenic and social aspects. The factors of exogenous origin determine development of adaptive changes. The present article summarises the mechanisms of animal venom toxins and homeostasis disruption in the body of mammals. The mechanisms underlying pathological changes are associated with shifts in biochemical reactions. Components of the immune, nervous and endocrine systems are key in the host defense and adaptation processes in response to venom by triggering signalling pathways (PI3kinase pathway, arachidonic acid cascade). Animal venom toxins initiate the development of inflammatory processes, the synthesis of pro-inflammatory mediators (cytokines), ROS, proteolytic enzymes, activate the migration of leukocytes and macrophages. Keratinocytes and endothelial cells act as protective barriers under the action of animal venom toxins on the body of mammals. In addition, the formation of pores in cell membranes, structural changes in cell ion channels are characteristic of the action of animal venom toxins.

EFFECT OF SCORPION VENOM TOXINS ON STRUCTURAL AND FUNCTIONAL PARAMETERS OF INTERNAL ORGANS, INCLUDING KIDNEYS (REVIEW).

OBJECTIVE: The aim: To establish patterns of structural and functional changes in internal organs, including kidneys, under the conditions of exposure to scorpion venom toxins. PATIENTS AND METHODS: Materials and methods: A thorough literature analysis was conducted on the basis of PubMed, Google Scholar, Web of Science, and Scopus databases. When processing the search results, we chose the newest publications up to 5 years old or the most thorough publications that vividly described the essence of our topic. CONCLUSION: Conclusions: The venom of various species of scorpions exhibits a wide range of biological activity. Acting on the structures of the central and peripheral nervous system, the toxins of scorpion venom cause the development of paralysis, convulsions, brain inflammation, hemorrhagic and ischemic strokes. Under conditions of influence on the cardiovascular system, damage to the endothelial lining of the vascular wall, disturbances in heart rhythm, conduction, and the development of destructive changes in the myocardium are characteristic. Data on kidney damage due to scorpion bites require a more detailed study, as information on microscopic and submicroscopic changes in the structure of the organ is too limited. However, cases of the development of tubular necrosis, interstitial nephritis, and kidney infarction are currently known.

Protein and peptide profiles of rats' organs in scorpion envenomation.

Problem of scorpion envenomation becomes more alarming each year. Main effects of scorpion venom are commonly believed to be related to its neurotoxic properties, yet severe symptoms may also be developed due to the uncontrolled enzymatic activity and formation of various bioactive molecules, including middle-mass molecules (MMMs). MMMs are considered as endogenous intoxication markers, their presence may indicate multiple organ failure. Scorpions, belong to the Leiurus macroctenus species, are very dangerous, nevertheless, effects of their venom on protein and peptide composition within the tissues remains unclear. In this work we have focused the attention on changes in protein and MMM levels and peptide composition in various organs during Leiurus macroctenus envenomation. The results revealed a decrease in protein level during envenomation as well as a significant increment of MMM210 and MMM254 levels in all assessed organs. Quantitative and qualitative compositions of various protein and peptide factions were continually changing. All of this may suggest that Leiurus macroctenus sting causes considerable destruction of cell microenvironment across all essential organs, providing systemic envenomation. In addition, MMM level increment may indicate endogenous intoxication development. Peptides, formed during envenomation, may possess various bioactive properties, analysis of which constitutes an area of further studies.

The Activity of Metalloproteases and Serine Proteases in Various Organs after Leiurus macroctenus Envenomation.

Background: Scorpion stings may be life-threatening since their venoms are comprised of a wide range of toxins and other bioactive molecules, such as enzymes. At the same time, scorpion envenomation may increase matrix metalloproteases (MMPs) levels, which enhance proteolytic tissue destruction by venom. However, investigations on the impact of many scorpions' venoms, such as those of Leiurus macroctenus, on tissue proteolytic activity and MMP levels have not yet been conducted. Methods and Results: The present study aimed to examine the total proteolysis levels in various organs after Leiurus macroctenus envenomation and evaluate the metalloproteases and serine proteases' contributions to the total proteolytic activity. Changes in MMPs and TIMP-1 levels were tested as well. Envenomation led to a significant increase in proteolytic activity levels in all assessed organs, mostly in the heart (by 3.34 times) and lungs (by 2.25 times). Conclusions: Since EDTA presence showed a noticeable decrease in total proteolytic activity level, metalloproteases appeared to play a prominent role in total proteolytic activity. At the same time, MMPs and TIMP-1 levels were increased in all assessed organs, suggesting that Leiurus macroctenus envenomation causes systemic envenomation, which may induce multiple organ abnormalities, mostly because of the uncontrolled metalloprotease activity.

PROTEOLYTIC ACTIVITY IN THE HEART OF RATS WITH HYPERHOMOCYSTEINEMIA.

OBJECTIVE: The aim: To investigate the distribution of proteolytic activity and cytokine profile in the heart of rats with hyperhomocysteinemia. PATIENTS AND METHODS: Materials and methods: A total of 60 albino non-linear male rats was used in the study. Hyperhomocysteinemia was induced by intragastric administration of DL-homocysteine thiolactone. Total proteolytic activity was measured using casein as a substrate. To determine the activity of metal-dependent and serine proteases, ethylenediaminetetraacetic acid, and phenylmethylsulfonyl fluoride were used. The level of matrix metalloproteinases, tissue inhibitor of metalloproteinases-1, and cytokines was studied by enzyme-linked immunosorbent assay. RESULTS: Results: It was found an increase in the total proteolytic activity in the heart of young, adult, and old animals. In addition, the redistribution of proteolytic activity was revealed - the portion of metal-dependent enzymes increased in all groups while the percentage of serine proteases decreased in the old animals with hyperhomocysteinemia. The state of mild inflammation, evidenced by the increased level of some pro-inflammatory cytokines, was found in the heart of young and old animals with hyperhomocysteinemia. CONCLUSION: Conclusions: The pathogenesis of hyperhomocysteinemia is accompanied by a change in the proteolytic activity in the heart as well as a change in the cytokine profile.

Mechanisms of the Impact of Hashimoto Thyroiditis on Papillary Thyroid Carcinoma Progression: Relationship with the Tumor Immune Microenvironment.

BACKGROUND: The relationship between Hashimoto thyroiditis (HT) and papillary thyroid carcinoma (PTC) remains uncertain. We assessed the impact of HT on the tumor immune microenvironment (TIME) in PTC. METHODS: Thirty patients with PTC (group 1) and 30 patients with PTC and HT (group 2) were enrolled in this pilot study. The distribution and number of CD8+ lymphocytes, plasma cells (CD138+), regulatory T cells (forkhead box P3 [FOXP3+)], mast cell tryptase (MCT+), and M2 macrophages (CD163+) were evaluated. To test the hypothesis that HT impacts PTC development via signal transducer and activator of transcription 6 (STAT6) activation and M2 macrophage polarization, we investigated STAT6 expression in tumor and stromal cells. We also evaluated vascular endothelial growth factor (VEGF) expression by lymph node metastasis (LNM) status. RESULTS: TIME showed significant between-group differences. Group 1 patients demonstrated immune desert or immune-excluded immunophenotypes, while an inflamed phenotype with more CD8+ cells (P<0.001) predominated in group 2. Immune-excluded TIME was associated with the highest LNM rate. In PTC, LNM was associated with more numerous CD163+ cells. Moreover, LNM in group 1 was associated with increased numbers of mast cells peritumorally and FOXP3+ cells intratumorally and peritumorally. Group 2 demonstrated higher STAT6 but not higher VEGF expression in tumor cells. High VEGF expression was associated with LNM regardless of HT status. CONCLUSION: Concomitant HT impacted PTC signaling via STAT6 and TIME by increasing the number of CD8+ cells. LNM is associated with increases in CD163+ cells and VEGF expression in PTC, whereas HT affected LNM through different mechanisms.

Circadian Rhythms of Endothelial Nitric Oxide Synthase and Toll-like Receptors 2 Production in Females with Rheumatoid Arthritis Depending on NOS3 Gene Polymorphism.

BACKGROUND: Rheumatoid Arthritis (RA) is an autoimmune polygenic disease characterized by rapid disability progression and high prevalence. Progression of RA is closely associated with chronobiological changes in the production of some hormones and inflammatory mediators, influencing the disease course and therapy efficacy. The main pathogenetic mechanism of RA is angiogenesis, which is controlled by biological clock-genes. Further investigation of circadian rhythms of angiogenic mediators production in RA patients may be considered as important and relevant. The aim of this study was to establish daily variability of serum endothelial Nitric Oxide Synthase (NOS3) and toll-like receptors 2 (sTLR2) levels in female RA patients depending on the NOS3 gene polymorphism. METHODS: We examined 173 RA patients (100% female) aged 43.7 ± 7.35 years and 34 age-matched healthy women without joint diseases and autoimmune diseases (control). RA was diagnosed by ACR/EULAR 2010 criteria. Blood serum NOS3 and sTLR2 levels were determined at 08:00 and 20:00 using Cloud-Clone Corp kits (USA). NOS3 T-786С (rs2070744) polymorphism was determined by Real-Time PCR (Bio-Rad iCycler IQ5) using SNP-express kits. The SPSS22 software package was used for statistical processing of the results. RESULTS: Females with RA demonstrated oppositely directed serum NOS3 and sTLR2 daily changes: NOS3 level in the morning (08:00) was lower than in the evening (+ 45.5 ± 30.7%), and sTLR2 level in the evening (at 20:00) was lower than in the morning (-21.6 ± 13.1%). RA patients had differences in NOS3 and sTLR2 production depending on NOS3 T786C genotype. CC subjects had NOS3 level at 08:00, 20:00 and day average levels lower (16-25%), and sTLR2 level higher (24-27%) than those of TT subjects. RA patients, carriers of CC genotype, had higher chances of NOS3 and sTLR2 aberrant production compared to TT and TC genotype carriers (OR = 2.99 and 4.79, respectively). CONCLUSION: RA patients demonstrated oppositely directed circadian changes of serum NOS3 and sTLR2. CC genotype carriers had lower NOS3 and higher sTLR2 production rates than TT and TC genotype carriers.

The Role of Hyperhomocysteinemia in the Development of Changes in the Lungs.

BACKGROUND: In recent years, attention of the scientific community has been drawn to the study of the role of homocysteine in the pathology of diseases in general and the respiratory system in particular. Violations of the synthesis and disposal of homocysteine are the cause of its excessive accumulation in the body, which subsequently leads to damage to all organs and systems. METHODS: We conducted this study on 64 white non-linear rats of 6-8 and 24-26 months old rats, which were divided into two control and experimental groups during the experiment. The study of ultrastructural changes in the lungs of rats was performed using an electron microscope. RESULTS: The results of the conducted study show that all the animals of the experimental group, corresponding to 32 (50%) cases, possess conspicuous morphological changes in the structure of the lungs. The specific features of mature animals were the narrowing of the alveolar space of the part of the alveoli and the increase in the number of macrophages in them. Partial desquamation of capillary endothelial lining and a slight increase of collagen fibers in interalveolar membranes were observed compared to control animals. A characteristic feature for older animals was the growth of connective tissue, predominantly collagen fibers, which led to pneumosclerosis. The thickening of the aerogemic barrier was also detected, and the endothelial lining was intermittent or desquamated. CONCLUSION: Mature animals of the experimental group were characterized by activation of inflammatory processes, oxidative stress, due to increased production of macrophages and, as a consequence, the launch of the humorous link of immunity, while in old rats were determined by fibrosis, disorders of the trophic and gas metabolism, as well as damage to the endothelium part of lungs.