[The effect of vitamin K deficiency in rats on various enzyme systems participating in energy metabolism]. / Vliianie nedostatochnosti vitamina K u krys na nekotorye fermentnye sistemy, uchastvuiushchie v énergoobespechenii.

In alimentary deficiency of vitamin K in rats, accompanied by an increase in the prothrombin time by 30%, activity of kidney creatine kinase and of blood serum alkaline phosphatase was unaltered, while the activity of alkaline phosphatase in small intestinal mucose was decreased by 20% and that of creatine kinase from skeletal muscles--by 10%. In vitamin K-deprived animals the rate of coupling between respiration and mitochondrial phosphorylation was decreased, which might be due to alteration in the NADH-dehydrogenase complex. Menadion reductase activity and cyanide-resistant respiration of mitochondria were unaltered in presence of menadion. Palmitic acid effectively activated of mitochondrial respiration in vitamin K-deprived animals (contrary to the control rats). This effect appears to occur as a result of structural alterations in mitochondria depending on vitamin K level in the organelles.

[Use of immunospecific methods for quantitative estimation of microsomal epoxide hydrolase]. / Ispol'zovanie immunospetsificheskikh metodov dlia opredeleniia kolichestva mikrosomal'noi époksidgidrolazy.

Antiserum, specific to rat liver microsomal epoxide hydrolase, was produced. Three methods for quantitative estimation of the epoxide hydrolase, base on the principle of antigen-antiserum immunoprecipitation, were used: electroimmune analysis by Laurell, radial immunodiffusion, enzyme-linked immunoassay. Distinct correlation between specific enzyme activity and its content in microsomal preparation was demonstrated using a simple and stable procedure of radial immunodiffusion. In the enzyme-linked assay conjugate of protein A with alkaline phosphate served as the secondary antibody. Sensitivity of the procedure described was about 1,000-fold higher as compared with radial immunodiffusion method.

[Experimental evaluation of the effectiveness of vitamin-enriched chopped meat products]. / Eksperimental'naia otsenka éffektivnosti vitaminizirovannykh miasnykh rublenykh izdelii.

The ration devoid of vitamins B1 and B2 and niacin, given to growing rats during 4-10 weeks led to growth cessation, a decrease in body mass and the development of vitamin deficiency that was expressed in a drastic lowering of red blood cell transketolase activity, and to increased effects of TDF and FAD. Inclusion into the animals' ration of cutlets enriched with vitamins B1, B2, C and niacin completely normalized their growth and biochemical parameters of their providing with vitamins B1 and B2. Routine cutlets were less effective. The feeding of rats with vitamin-enriched cutlets during 10 weeks did not induce any histological or histochemical disorders in their internal organs.

[Enzymatic activity of the microsomal fraction of the mucosa of the small intestine in guinea pigs with vitamin D and C deficiencies]. / Aktivnost' nekotorykh fermentov mikrosomal'noi fraktsii slizistoi obolochki tonkoi kishki morskikh svinok pri nedostatochnosti vitaminov D i C.

The influence of vitamin D and C deficiency on the kinetic parameters of sucrase and alkali phosphatase activities was studied in the microsomal fraction of the small intestinal mucosa of guinea pigs. It was found that Km values for these enzymes did not depend on the animal providing with these vitamins. Deficiency of one of these vitamins did not influence sucrase activity, however, simultaneous elimination of vitamins D and C resulted in the activity rise by 92%. Alkali phosphatase and Ca-ATPase activities proved to be similarly dependent on providing with vitamin D in the presence of vitamin C in the ration, while in the absence of vitamin C this dependence was not observed.

[Vitamin D and calcium metabolism in relation to different levels of vitamins B6 and D]. / Obmen vitamina D i kal'tsiia pri razlichnoi obespechennosti vitamina B6 i D.

The influence of vitamin B6 deficiency on some vitamin D-dependent processes was studied in animals. The following parameters changing in relation to the level of vitamin D providing were investigated: activity of alkaline phosphatase in the serum and small intestine mucosa, the levels of Ca, P and parathormone, concentration of vitamin D metabolites and enzyme activity; and only 25-hydroxyvitamin D (25-OVD) concentration in the blood serum, under conditions of combined vitamin B6 and D deficiency was significantly lower as compared to cases with vitamin D deficiency alone. In the presence of vitamin B6 deficiency recovery of 25-OVD level in the blood serum, after vitamin D administration to the animals, had a tendency to delay as compared to that in the animals provided with vitamin B6. Vitamin B6 deficiency produced similar effect on 25-OVD 1-hydroxylase activity. The data obtained have evidenced a possibility of vitamin B6 influence on vitamin D metabolism.

Comparative evaluation of the depth of collagen and hyaluronic acid hydrolysis in vitro by collagenase and hyaluronidase preparations.

The molecular weights of collagen and hyaluronic acid solutions after their incubation with collagenase and hyaluronidase were evaluated by capillary viscosimetry. The results indicate high amylolytic activity of collagenase and the absence of proteolytic activity in hyaluronidase in an in vitro system.

Dynamics of hydration of cicatricial tissue during local therapy with collagenase preparations.

The components of complex dielectric permeability of cicatricial tissue at the frequency of 55 GHz were studied during local therapy of cicatrices with Fermenkol (a complex of collagenolytic proteases). Electrical characteristics of tissues in the microwave range were interpreted in terms of hydration parameters (total content of water in tissues and the ratio of structured and free water). The rightfulness of interpretations is discussed on the basis of the results of measurements of cell suspension models (native blood and its fractions brought to a standard hematocrit). The results attest to a relationship between hydration of the cicatricial tissue and its morphology and function.

Role of sex hormones in cardiovascular resistance to atropine in rats at rest and during stress.

Castration of males and females reduces the sensitivity of cardiac chronotropic function to atropine and potentiates the vascular component in the reaction to atropine in females (during stress) and males (at rest and during stress). Our results show that estrogens and androgens increase the sensitivity of the cardiovascular system to cholinergic influences at rest and during stress.

The preparation of soft-tritium-labelled proteins and their application for the collagenolytic activity investigations.

Tritium-labelled Type I collagen from rat skin with specific activity of 8 mCi/mg and mixture of the crab hepatopancrease collagenolytic enzymes with specific activity of 25 mCi/mg have been obtained by the method of high-temperature solid-state catalytic isotope exchange (HSCIE) without loss of native features. Tritium-labelled Type I collagen kept its ability to form native fibrils being resistant to trypsin hydrolysis. Tritium-labelled collagenase keeps about 70% of its proteolytic activity.

Collagenolytic protease preparations from invertebrates: biochemical aspects of medical and cosmetological applications.

Collagenolytic proteases from invertebrates, the active matter of new structure-destroying enzyme preparations (Collagenase from hydrobionts, Polycollagenase-K, Fermenkol), ensure deep hydrolysis of polypeptide substrates (native or partially denatured collagen types, elastin, fibrin, hemoglobin, and casein) that cannot be attained by collagenases and serine proteases. Biochemical properties of collagenolytic proteases from invertebrates, techniques and doses that may be important for elaboration and employment of preparations based on this complex are described.

Effects of collagenolytic protease preparations from invertebrates on the immune system.

We studied the effects of an active ingredient of new enzyme preparations containing collagenolytic proteases from various hydrobionts (Collagenase from hydrobionts, Polycollagenase-K, and Fermenkol). These substances in therapeutic doses did not affect the humoral immune response estimated by hemagglutination and local hemolysis in gel and caused no local irritation and allergic reactions after long-term application to the skin.

Collagenolytic protease preparations from invertebrates: experimental study of acute and chronic toxicity.

Toxicity of active matter of new structure-destroying enzyme preparations containing a complex of collagenolytic proteases from invertebrates (Collagenase from hydrobionts, Polycollagenase-K, Fermenkol) was studied. The absence of acute toxicity in skin application is proved, and LD(50) for intraperitoneal administration to mice and rats were determined. A long-term treatment with clinical doses under various types of application does not affect body weight gain, biochemical indices of the plasma, ECG, diuresis, and urine content.

The isolation and properties of collagenolytic proteases from crab hepatopancreas.

A mixture of collagenolytic proteases has been isolated from the Kamchatka crab hepatopancreas. The four individual enzymes were further separated with FPLC and partially characterized. Crab collagenolytic proteases possess a high activity against different types of collagen, especially against calf skin collagen Type III and bovine lens capsule collagen Type IV, which is resistant to the microbial Clostridium sp. collagenases. In contrast with microbial collagenases the crab enzymes are good general proteases, able to cleave standard synthetic and protein substrates and possess a chymotrypsin-, trypsin- and elastase-like specificity. N-Terminal sequence analysis revealed that crab collagenolytic proteases had evolved from a trypsin-like ancestor. Crab proteases, structurally belonging to the trypsin-like enzymes, nevertheless, possess the unique ability, among this class of enzymes, to cleave the native insoluble collagen. It seems that crab collagenolytic proteases and true metalloenzyme vertebrate and microbial collagenases have certain common structural features particularly in the regions of their substrate binding site.