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.

Low-Molecular-Mass Fragments of Collagen Improve Parameters Related to Mass and Inflammation of the Adipose Tissue in the Obese Rat.

Research background: Despite clearly recognized links between increased body mass and increased risk for various pathological conditions, therapeutic options to treat obesity are still very limited. The aim of the present study is to explore the effect of low-molecular-mass collagen fragments obtained from the scales of Antarctic wild marine fish on rats' visceral and subcutaneous white adipose tissue in a high-calorie diet-induced obesity model. Experimental approach: The study was conducted on outbred rats, which were divided into 3 experimental groups: (i) control, consuming standard food (3.81 kcal/g), (ii) obese group, consuming a high-calorie diet (5.35 kcal/g), and (iii) obese group, consuming a high-calorie diet (5.35 kcal/g) with intragastric administration of low-molecular-mass collagen fragments (at a dose 1 g/kg of body mass during 6 weeks). Low-molecular-mass collagen fragments were obtained by a procedure that included collagen extraction from fish scales and enzymatic hydrolysis with pepsin. Apart from hematoxylin and eosin staining, fibrosis level was assessed by histochemical Van Gieson's trichrome picrofuchsin staining, and mast cells were analysed by toluidine blue O staining. Results and conclusions: Group treated with low-molecular-mass fragments of collagen exhibited decreased rate of mass gain, relative mass, area occupied by collagen fibre of both visceral and subcutaneous adipose tissue, and cross-sectional area of both visceral and subcutaneous adipocytes. Treatment with low-molecular-mass fragments of collagen reduced the infiltration of immune cells, number of mast cells and their redistribution back to the septa. This was also accompanied by a decreased number of the crown-like structures formed by the immune cells, which are markers of chronic inflammation that accompanies obesity. Novelty and scientific contribution: This is the first study that reports the anti-obesity effect of low-molecular-mass fragments produced as a result of controlled hydrolysis of collagen from the scales of Antarctic wild marine fish in the in vivo model. Another novelty of this work is the observation that the tested collagen fragments not only reduce the body mass, but also improve the morphological and inflammatory parameters (decrease in the number of crown-like structures, immune cell infiltration, fibrosis and mast cells). Altogether, our work suggests that low-molecular-mass collagen fragments are a promising candidate for amelioration of some comorbidities linked to obesity.

Anti-obesity effect of collagen peptides obtained from Diplulmaris antarctica, a jellyfish of the Antarctic region.

AIM: To investigate the ability of collagen peptides derived from a jellyfish of the Antarctic region (Diplulmaris antarctica) to prevent the development of obesity in rats fed a high-calorie diet. METHODS: Collagen peptides were produced by pepsin hydrolysis of jellyfish-derived collagen. The purity of collagen and collagen peptides was confirmed by SDS-polyacrylamide gel electrophoresis. Rats were fed a high-calorie diet for ten weeks and were simultaneously orally administered collagen peptides (1 g per 1 kg of body weight every other day) starting from the fourth week. Body mass index (BMI), body weight gain, selected nutritional parameters, the key parameters associated with insulin resistance, and the level of oxidative stress markers were assessed. RESULTS: Compared with untreated obese rats, rats treated with hydrolyzed jellyfish collagen peptides had a decreased body weight gain and body mass index. They also had a decreased level of fasting blood glucose, glycated hemoglobin, insulin, lipid peroxidation products (conjugated dienes, Schiff bases), and oxidatively modified proteins, as well as a restored activity of superoxide dismutase. CONCLUSION: Collagen peptides obtained from Diplulmaris antarctica can be used to prevent and treat obesity caused by a high-calorie diet and pathologies associated with increased oxidative stress. Given the obtained results and the abundance of Diplulmaris antarctica in the Antarctic region, this species can be considered a sustainable source of collagen and its derivatives.

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.

The Rise of Factor X Level in Blood Plasma of Patients at Severe Burn Injuries.

This work is dedicated to the detection of imbalance between the pro- and anticoagulant branches of hemostasis at severe burn injuries by evaluating the content or activity of individual clotting factors. To select the targets for accurate diagnostics we measured the concentrations of soluble fibrin monomeric complexes and fibrinogen, levels of total prothrombin, factor X, protein C, and antithrombin III, and recorded the time of clotting in activated partial thromboplastin time and prothrombin time (PT) tests. Factor X level was increased in 26% of patients on the 1st day after the burn and it rose further in 62% patients on the 14th day of recovery. Increasing factor X level is assumed to be a risk factor of thrombotic complications. We propose to use it as a marker of predisposition to thrombosis at severe burn injury.

Dual soluble epoxide hydrolase inhibitor/PPAR-γ agonist attenuates renal fibrosis.

Renal fibrosis is a contributor to chronic kidney disease and an important predictor of long-term prognosis. We developed a dual soluble epoxide hydrolase inhibitor-PPAR-γ agonist (sEHi/PPAR-γ), RB394, and investigated its ability to attenuate renal fibrosis in a mouse unilateral ureteral obstruction (UUO) model. RB394 efficacy was compared to an sEH inhibitor (sEHi), a PPAR-γ agonist rosiglitazone (Rosi), or their combination (sEHi + Rosi). All interventional treatments were administrated in drinking water 3 days after UUO induction surgery and continued for 7 days. UUO mice developed renal fibrosis with higher collagen formation and RB394 significantly attenuated fibrosis (P < 0.05). Renal expression of α-smooth muscle actin (α-SMA) was elevated in UUO mice and all treatments except sEHi significantly attenuated renal α-SMA expression. Renal mRNA expression fibrotic and fibrosis regulators were higher in UUO mice and RB394 and sEHi + Rosi treatments attenuated their expression. Renal inflammation was evident in UUO mice with increased infiltration of CD45 and F4/80 positive cells. RB394 and sEHi + Rosi treatments attenuated renal inflammation in UUO mice. UUO mice had renal tubular and vascular injury. Renal tubular and vascular injuries were attenuated to a greater extent by RB394 and sEHi + Rosi than sEHi or Rosi treatment alone. Renal mRNA expression of oxidative stress markers were significantly higher in UUO mice (P < 0.05). RB394 and sEHi + Rosi attenuated expression of oxidative stress markers to a greater extent than other interventional treatments (P < 0.05). These findings demonstrate that RB394 can attenuate renal fibrosis by reducing renal inflammation, oxidative stress, tubular injury, and vascular injury. In conclusion, RB394 demonstrates exciting potential as a therapeutic for renal fibrosis and chronic kidney disease.

Hepatic Encephalopathy Aggravated by Systemic Inflammation.

BACKGROUND: The pathogenesis of hepatic encephalopathy (HE) is only partially understood. Beside ammonia accumulation in brain, a proinflammatory component has been suggested as precipitating event. OBJECTIVES: To evaluate the role of cytokines in cirrhosis for development of HE. METHODS: Pro- and anti-inflammatory cytokine profiles were determined in rats with CCL4-induced cirrhosis and HE as well as in patients with cirrhosis either due to metabolic disorders or chronic hepatitis C virus (HCV) with various grades of concomitant HE and depression. RESULTS: In the rat model and human cirrhosis a proinflammatory cytokine pattern (elevation of interferon gamma, interleukin [IL]-1ß, IL-6) was registered which in humans correlated to the degree of HE and depression. The most prominent elevation of proinflammatory cytokines was observed in chronic HCV as an additional inflammatory stimulus. In all cases of cirrhosis a comparable background activation of anti-inflammatory cytokines (e.g., IL-4) was detected which was interpreted as a physiologic counterbalance mechanism. CONCLUSIONS: The degree of HE and depression correlated with a proinflammatory cytokine pattern. It suggests that beside ammonia elevation, inflammatory cytokines determine occurrence and severity of hepatic encephalopathies. Thus, it can be defined a preferential therapeutic target.

The influence of immunoglobulin class G from blood plasma of patients with stroke on the activity of some parameters of hemostasis system.

Today, cardiovascular diseases are one of the main causes of disability of the population. Most of the illnesses, including stroke, are accompanied by the appearance immunoglobulin G (IgG) in the blood circulation. According to the literature sources and previous experiments, it is known that IgG made influence on the hemostasis system. Objectives of the investigation are pure enzymes: thrombin, factor X, proenzyme of protein C and prothrombin under the influence of IgG fraction that were separated from the plasma of patients with atherothrombotic and cardioembolic ischemic stroke and patients with other neurological diseases. IgG was separated by affinity chromatography. Disc electrophoresis was used to control antibodies' purity. The main goal of the experiment is to test the potential influence of all fractions of separated IgG on the process of hydrolysis of specific chromogenic substrates by the key factors of the hemostasis system such as thrombin, factor X and protein C. The appearance of IgG in the blood stream during the atherothrombotic and cardioembolic ischemic stroke and other neurological diseases was proved. Concentration of the IgG in plasma of patients with cardioembolic ischemic stroke was significantly higher compared with healthy donors, whereas the IgG in plasma of patients with atherothrombotic ischemic stroke and other neurological disorders was not significantly different. Their potential ability to influence the enzymes such as thrombin, factor X, proenzyme of protein C and prothrombin was shown. It was proved that the antibody fractions of all experimental groups significantly accelerate the process of splitting chromogenic substrate by thrombin but inhibited cleavage of a specific chromogenic substrate by protein C.

Synthesis and Anti-Platelet Activity of Thiosulfonate Derivatives Containing a Quinone Moiety.

Thiosulfonate derivatives based on quinones were synthesized for studying "structure-activity relationship" compounds with an acylated and a free amino-group. Anti-platelet activity of the synthesized compounds was determined and the influence of substituents on the activity of the derivatives was assessed.