Effect of tocopherol on biochemical blood parameters in pleuritis-induced rats treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin.

The aim of this study was to evaluate the effect of tocopherol on pleuritis-induced rats exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Rats were treated with a single TCDD dose of 5 µg/kg body weight (b.w.) and then for 3 weeks they were daily supplemented with tocopherol at a dose of 30 mg/kg b.w. The inflammation was initiated by intrapleural injection of a single dose of 1% carrageenin solution in a volume of 0.15 ml. Changes in biochemical blood parameters were measured three times at the 24th, 72nd and 120th hour of pleuritis and the blood was collected from 20 animals of each group of rats (group with the control inflammation; group treated with TCDD and with control inflammation; group treated with TCDD, supplemented with tocopherol and with the inflammation). The following biochemical parameters were measured: tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-2, IL-4, IL-6, procollagen, telopeptide, fibrinogen, cholesterol, urea, creatinine, aspartate aminotransferase (AspAT) and alanine aminotransferase (AlAT). Daily supplementation of tocopherol caused significant changes in the level of TNF, IL-1, IL-4, IL-6, urea, creatinine, AspAT and AlAT. According to the results of these studies, we suggest that tocopherol supplementation in high doses could act as a protective treatment to improve liver metabolism.

Influence of dioxin intoxication on the human system and possibilities of limiting its negative effects on the environment and living organisms.

INTRODUCTION AND OBJECTIVE: Despite the restrictive legal regulations related to the reduction of dioxins emission, their concentration in the environment is still too high. Mainly, this is related to the illegal utilisation of electronic equipment and combustion of wastes, and also to intensified activity and maintenance of ships, especially in developing countries. The most important remaining source in Europe is the metal industry. Studies on the mechanism of impact of dioxins are still being carried out. This review points at new possibilities for limiting the molecular mechanisms of dioxins activity, inter alia, through the application of high doses of tocopherol and acetylsalicylic acid while treating dioxins intoxication. BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: Apart from the knowledge of dioxins affinity to the aryl hydrocarbon receptor (AhR), the multi-stage radical-form actions and the pro-inflammatory mechanism associated with cyclooxygenase-II enzyme (COX-2) are under intense investigation at the moment. Due to the high affinity of dioxins to animals adipose tissue and their ability to accumulate in it, they can enter the food chain. Furthermore, high dioxin doses can cause poisoning manifested as advanced clinical symptoms, whereas in smaller doses, when cumulated, can cause metabolic changes which are often difficult to associate with their presence. Recently, some serious food contaminations by dioxins have been demonstrated. Sea fish and products from contaminated aqueducts still constitute potential sources of dioxins pollution. CONCLUSION: According to recent studies, dioxins are present in different concentrations in the environment and cause specific and long-time effects. These effects could be limited by the use of tocopherol and acetylsalicylic acid.

The influence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on hematological parameters during experimentally induced pleuritis in rats.

Proper functioning of homeostatic mechanisms is characteristic for every healthy organism and enables adapting to environmental changes. These complicated systematic reactions can neutralize the harmful stress factors leading to various inflammatory reactions. The aim of this study was to determine dynamic changes in the inflammatory reaction after single 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) administration of 5 µg/kg body weight into rats with experimentally induced pleuritis. These changes were observed by monitoring the hematological blood parameters during inflammation. The obtained results proved that dioxins contribute to various changes in the character of the inflammatory response. TCDD administration before pleuritis initiation caused an increase of lymphocytes and significant decrease of the number of neutrophils during inflammation. The current study proved that administration of low TCDD dose (seven times lower than used in other studies) can cause thymus, spleen, or lymphatic gland atrophy. This finding indicates the toxic influence of small TCDD dose especially on the immune system.

Mechlorethamine (NTG) effects on the erythrocytic and leukocytic blood parameters during experimentally induced pleuritis in rats.

BACKGROUND: According to cytotoxic and mutagenic properties, nitrogranulogen (NTG) changes the character of inflammatory reactions. Our previous studies have shown that NTG can enhance immunological defense reactions, because of its high affinity to DNA, and causes disorders in the synthesis of acute phase proteins (e.g., haptoglobin, transferrin, fibrinogen and complement protein C3) [15]. The aim of the current studies was to determine the influence of three different NTG doses: 5 µg/kg b.w. (body weight), 50 µg/kg b.w. and 600 µg/kg b.w. (cytotoxic dose) on the values of hematological blood parameters: RBC, HGB, HCT, RDW, MCV, MCH, MCHC, PLT, MPV, PCT, PDW, WBC, NEUT, LYMPH, MONO, EOS and BASO in pleuritis-induced rats. METHODS: The animals were randomized into five groups: Group I - control group; Group II - IP (induced pleuritis) group; Group III - NTG5 group; Group IV - NTG50 group; Group V - NTG600 group. The blood was collected from all the groups at the 24(th) h, 48(th) h, and 72(nd) h after the initiation of the carrageenin-induced inflammatory reaction. RESULTS: These investigations have revealed that NTG administered at the dose of 5 µg/kg b.w. caused the drop of the leukocyte and lymphocyte numbers and the rise of the neutrophil number at the 72(nd) h of the experimental-induced inflammatory reaction. Moreover, the dose of: 5 µg/kg b.w. was an immunomodulatory property and it also increased the erythrocytic parameters. On the contrary, NTG applied at the doses of 50 µg/kg and 600 µg/kg b.w. contributed to the drop of both: the erythrocytic and leukocytic parameters during the whole time of the inflammatory reaction. CONCLUSIONS: The results suggest that nitrogranulogen affects the erythropoiesis.

Synthesis of glycinated glycoconjugates based on 1-thioglycosides and their preliminary studies as potential immunomodulatory factor.

The biological importance of lipopolysaccharides (LPS), components of bacterial cell wall has not been explained sufficiently. The glycine present in these structures could play an important role in the immunological response after bacterial infections and during sepsis. In our studies we obtained synthetic and stable substituted glycinated 1-thioglycosides derivatives of monosaccharides, e.g., D-glucose or D-galactose as well as disaccharides, e.g., melibiose and lactose. The conditions of acylation reactions were validated and specific products were separated by using chromatography methods. Their structures were confirmed by NMR. These compounds were conjugated with carrier proteins e.g., bovine serum albumin and horse myoglobin. Prior to conjugation proteins were modified with glycidol to create the protein-diol intermediates and subsequent periodate oxidation of the glycol moieties to generate the reactive aldehyde functionalities. Modified and formylated carrier proteins were conjugated with acylated thioglycosides in the presence of sodium cyanoborohydride. Subsequently, the products obtained were analyzed in SDS-PAGE and separated by using HW-55S gel-filtration chromatography. The immunoreactivity of selected glycinated glycoconjugates were studied in ELISA assays with specific anti-aminoacylated glyconjugate antibodies obtained after rabbit immunization with Escherichia coli K12 C600 core oligosaccharide glycine-containing glycoconjugate. The differences in the immunoreactivity of different glycinated 1-thioglycosides were observed. The received glycine-acylated glycoconjugates could mimic the non-sugar substituents localized in various bacterial LPS. These synthetic compounds could be candidates for their use as glycoconjugate vaccines in protection against serious bacterial infections, e.g.. sepsis.

O-aminoacylation of bacterial glycoconjugates: from native structure to vaccine design.

The aminoacylation of bacterial polysaccharide antigens and its biological role are poorly understood, although it might be relevant in infection and immunity. Due to the lability of ester-linked substituents on glycoconjugate antigens, such groups usually escape detection during routine structural investigation. Of the few data available, those on the occurrence of glycine in the endotoxic lipopolysaccharides of Gram-negative bacteria are well documented. This article summarizes these data on glycine as an integral constituent of bacterial LPS and also some other amino-acid esters in the teichoic acids and phosphatidylglycerol of Gram-positive bacteria. The possible functions of such noncarbohydrate ester-linked substituents in bacterial antigens are discussed. Because glycine, an inherent component of bacterial lipopolysaccharides in the core region, is supposed to participate in epitope formation, such a structure may be considered for potential use in the construction of a vaccine with broad specificity.

Estimation of the action of three different mechlorethamine doses on biochemical parameters during experimentally induced pleuritis in rats.

Nitrogranulogen (NTG) may modify the character of inflammatory reactions. These modifications are a result of cytotoxic and mutagenic effects. NTG has high affinity to DNA and causes disorders in the synthesis of acute phase proteins (e.g., haptoglobin, transferrin, fibrinogen, and complement protein C3). Our previous studies have shown that small doses of NTG can enhance immunological defense reactions in the organism. The aim of the current studies was to determine how different NTG doses cause changes in the values of biochemical parameters in pleuritis-induced rats. The animals were randomized into five groups: Group I - control group; Group II - IP (induced pleuritis) group; Group III - NTG5 group; Group IV - NTG50 group; Group V - NTG600 group. Blood was collected from all groups of animals at 24, 48, and 72 h after the initiation of the carrageenin-induced inflammatory reaction. These investigations revealed that a dose of 5 µg NTG/kg b.w. (body weight) can change the character of the inflammation. Our studies also show that a dose of 600 µg NTG/kg b.w. causes a rapid decrease in the level of C3 at the 72 h of the experiment (after 3 applications every 24 h), which indicates a cytotoxic action of such a large NTG dose. NTG used at doses of 50 and 600 µg/kg b.w. causes the opposite metabolism of albumins and other serum proteins. Our studies show that the different doses of NTG have distinct effects on the inflammatory reaction.

Glycation of the muscle-specific enolase by reactive carbonyls: effect of temperature and the protection role of carnosine, pyridoxamine and phosphatidylserine.

Reactive carbonyls such as 4-hydroxy-2-nonenal (4-HNE), trans-2-nonenal (T2 N), acrolein (ACR) can react readily with nucleophilic protein sites forming of advanced glycation end-products (AGE). In this study, the human and pig muscle-specific enolase was used as a protein model for in vitro modification by 4-HNE, T2 N and ACR. While the human enolase interaction with reactive α-oxoaldehyde methylglyoxal (MOG) was demonstrated previously, the effect of 4-HNE, T2N and ACR has not been identified yet. Altering in catalytic function were observed after the enzyme incubation with these active compounds for 1-24 h at 25, 37 and 45 °C. The inhibition degree of enolase activity occurred in following order: 4-HNE > ACR > MOG > T2N and inactivation of pig muscle-specific enolase was more effective relatively to human enzyme. The efficiency of AGE formation depends on time and incubation temperature with glycating agent. More amounts of insoluble AGE were formed at 45 °C. We found that pyridoxamine and natural dipeptide carnosine counteracted AGE formation and protected enolase against the total loss of catalytic activity. Moreover, we demonstrated for the first time that phosphatidylserine may significantly protect enolase against decrease of catalytic activity in spite of AGE production.

[The composition, biochemical properties and toxicity of snake venoms]. / Sklad i wlasciwosci biochemiczne oraz toksycznosc jadów wezy.

2.5 million cases of snake bites are noticed in the world every year (within 100,000 is mortal). These bites occur frequently in Asia and Africa. Some reports proved the toxicity and composition changes of well-known venoms from the same snake species according to the climatic zone. Snake venom is a natural source of many biologically active substances, including those with potential therapeutic properties. These substances contain peptides, proteins, and enzymes which are divided into five subfamilies: three-finger toxins, serine protease inhibitors of the Kunitz type, phospholipases A2, serine proteases, and metalloproteases. All snake venoms are grouped depending on their mode of action. They usually cause neurotransmission disorders, cardiotoxic action, hemostasis disorders, and have central nervous system and necrotic activity.

[Amphibian skin secretions as a new source of antibiotics and biologically active substances]. / Substancje wydzielane przez gruczoly skórne plazów jako nowe zródlo antybiotyków i zwiazków biologicznie czynnych.

So far, the main sources of biologically active substances used in medicine have been plants, molds, and propolis. The obtained compounds have either therapeutic features or require additional modification. They are sometimes combined with other pharmacological substances to intensify their therapeutic effect. However, the effectiveness of many drugs has been rapidly decreasing.The overuse of antibiotics in the treatment and prophylaxis of human infections (especially in hospitals) as well as their widespread and often unjustified use in the treatment and prophylaxis of farm animal illnesses contribute to the development of a variety of resistance mechanisms by microorganisms. Because of the increasing ineffectiveness of antibiotics used so far and difficulties in obtaining new drugs, it is necessary to find new sources of these compounds, for example in animal organisms. Research has demonstrated that amphibian skin secretions are rich in a variety of active substances which have strong pharmacological properties. In these compounds we can distinguish, for example, toxins, antimicrobial peptides, opioid peptides, steroids, and alkaloids.These compounds show cytotoxic, antimicrobial, analgesic, anti-inflammatory, and even antiviral activities (including anti-HIV). These substances can be used in cell receptor studies and in transmembrane ion transport analysis. Because these compounds are secreted by skin glands,they can be easy obtained without injuring these animals. It is probable that amphibian skin constitutes a potential source of modern drugs.

[Characterization of an inflammatory response]. / Charakterystyka odczynu zapalnego.

The intensity of an inflammatory response in a tissue or an organ is dependent on the efficiency of the organism's homeostatic mechanisms, which restrict the extent of the reaction. The type of factor inducing a inflammatory response and its strength have significant influence on the dynamics of an inflammatory reaction. The prompt eradication of an inflammatory factor and its biologically adverse effects attest to the efficacious adaptive mechanisms of the organism. The inflammatory response expresses biochemical, hematological, and immunological responses at the local or systemic level.