Development of Low-Molecular Weight Collagen Peptide Complex with Glycosaminoglycan Components.

Enzymatic hydrolysis of biopolymers of the cartilage tissue was studied for obtaining a complex of type II collagen peptides and glycosaminoglycan oligosaccharides. Hydrothermal hydrolysis in a high pressure homogenizer followed by enzymatic hydrolysis of the cartilage tissue biopolymers with proteolytic enzyme preparation Karipazim yielded a complex of collagen peptides and glycosaminoglycan oligosaccharides with molecular weights of 240-720 Da. Low molecular weight of the components increases their bioavailability. Entering into the cells (chondrocytes), low-molecular-weight peptides, disaccharides, and oligosaccharides as structural elements of the matrix can participate in the formation of fibrils of collagen and proteoglycans. Exogenous substances replenish deficient components of the matrix and/or their concentrations, affect the formation and strengthen the cartilage tissue. Thus, using cattle and porcine hyaline cartilages, we prepared a complex of biopolymers with lower molecular weights in comparison with previously developed nutraceuticals.

Metabolic correction by pyruvate halts acquired epilepsy in multiple rodent models.

Metabolic intervention strategy of epilepsy treatment has been gaining broader attention due to accumulated evidence that hypometabolism, manifested in humans as reduced brain glucose consumption, is a principal factor in acquired epilepsy. Therefore, targeting deficient energy metabolism may be an effective approach for treating epilepsy. To confront this pathology we utilized pyruvate, which besides being an anaplerotic mitochondrial fuel possesses a unique set of neuroprotective properties as it: (i) is a potent reactive oxygen species scavenger; (ii) abates overactivation of Poly [ADP-ribose] polymerase 1 (PARP-1); (iii) facilitates glutamate efflux from the brain; (iv) augments brain glycogen stores; (v) is anti-inflammatory; (vi) prevents neuronal hyperexcitability; and (vii) normalizes the cytosolic redox state. In vivo, chronic oral pyruvate administration completely abolished established epileptic phenotypes in three accepted and fundamentally different rodent acquired epilepsy models. Our study reports metabolic correction by pyruvate as a potentially highly effective treatment of acquired epilepsies.

[NMR study of human biological fluids for detection of pathologies].

The paper deals with the NMR spectra obtained using preparations of five different human biological body fluids. Characteristic metabolite signals of blood, urine, tears, saliva, and sweat spectra have been determined and classified. The biological body fluid samples were used for search and identification of biomarkers of cardiovascular disease. Absolute functional biomarkers for diseases such as coronary heart disease (CHD) have not been recognized even in the case acute myocardial infarction. A hypothesis explaining reasons of lack of such markers has been formulated. The results of comparative analysis of blood and urine samples from humans and some laboratory animals are given. Identify and analyze signals of metabolites of pathogenic microflora and their dynamics in the urine from patients with urogenital diseases have been determined and analyzed and characteristic biomarkers have been recognized.

Anxiolytic-like effects of AP7 injected into the dorsolateral or ventrolateral columns of the periaqueductal gray of rats.

RATIONALE: Glutamate antagonists microinjected into the dorsolateral PAG (DLPAG) show an anxiolytic-like profile in the elevated plus maze. Other columns of the PAG are also involved in defensive reactions. Few studies, however, have investigated the effects of pharmacological manipulation of the ventrolateral PAG (VLPAG) on procedures that predict anxiolytic activity. OBJECTIVES: To investigate the effects of the NMDA receptor (NMDAr) antagonist 2-amino-7-phosphonoheptanoic acid (AP7) microinjected into the DL or VLPAG in two procedures that predict anxiolytic activity using distinct aversive contingencies, the elevated plus maze and the Vogel punished licking test. METHODS: Male Wistar rats (7-14/group) with cannulas aimed at the DLPAG or VLPAG received AP7 (2 nmol/0.5 microl) or saline and 10 min later were submitted to the behavioural tests. In the punished licking experiment, water deprived (48 h) animals were allowed to drink for 3 min, receiving a 0.5 mA shock every 20 licks. The elevated plus maze test was performed as described elsewhere. Using this test, a dose response-curve for AP7 (0.2-20 nmol) injected in a smaller volume (0.25 microl) into the VLPAG was also performed. RESULTS: AP7 increased exploration of open arms of the EPM when microinjected into either the DLPAG or VLPAG ( P<0.05, ANOVA). The drug also increased the number of punished licks when administered into those columns (ANOVA, P<0.05). CONCLUSIONS: The results suggest that antagonism of endogenous excitatory amino acid neurotransmission in the DLPAG or VLPAG is able to reverse behavioral suppression induced by distinct aversive contingencies.

Defense reaction induced by a metabotropic glutamate receptor agonist microinjected into the dorsal periaqueductal gray of rats.

The behavioral effects of trans-(+/-)-1-amino-1, 3-cyclopentanedicarboxylic acid (t-ACPD), a metabotropic glutamate receptor (mGluR) agonist, or 0.9% (w/v) saline, injected into the dorsal periaqueductal gray (DPAG), was investigated. Male Wistar rats showed defense reactions characterized by jumps toward the top edges of the cages (saline = 0 vs t-ACPD = 6.0, medians P<0.05) and gallops (saline = 0 vs t-ACPD = 10.0, medians P<0.05) during the 60-s period after the beginning of the injection. In another experiment animals were placed inside an open arena for 5 min immediately after injection. Their behavior was recorded by a video camera and a computer program analyzed the videotapes. Eleven of fifteen rats injected with t-ACPD showed a short-lasting (about 1 min) flight reaction. No saline-treated animal showed this reaction (P<0.0005, chi-square test). The drug induced an increase in turning behavior (P = 0.002, MANOVA) and a decrease in the number of rearings (P<0.001, MANOVA) and grooming episodes (P<0.001, MANOVA). These results suggest that mGluRs play a role in the control of defense reactions in the DPAG.

Defense reaction induced by a metabotropic glutamate receptor agonist microinjected into the dorsal periaqueductal gray of rats

The behavioral effects of trans-(+ or -)-1-amino-1,3-cyclopentanedicarboxylic acid (t-ACPD), a metabotropic glutamate receptor (mGluR) agonist, or 0.9 per cent (w/v) saline, injected into the dorsal periaqueductal gray (DPAG), was investigated. Male Wistar rats showed defense reactions characterized by jumps toward the top edges of the cages (saline = 0 vs t-ACPD = 6.0, medians P<0.05) and gallops (saline = 0 vs t-ACPD = 10.0, medians P<0.05) during the 60-s period after the beginning of the injection. In another experiment animals were placed inside an open arena for 5 min immediately after injection. Their behavior was recorded by a video camera and a computer program analyzed the videotapes. Eleven of fifteen rats injected with t-ACPD showed a short-lasting (about 1 min) flight reaction. No saline-treated animal showed this reaction (P<0.0005, chi-square test). The drug induced an increase in turning behavior (P = 0.002, MANOVA) and a decrease in the number of rearings (P<0.001, MANOVA) and grooming episodes (P<0.001, MANOVA). These results suggest that mGluRs play a role in the control of defense reactions in the DPAG.

Ferroelectric active models of ion channels in biomembranes.

Ferroactive models of ion channels in the theory of biological membranes are presented. The main equations are derived and their possible solutions are shown. The estimates of some experimentally measured parameters are given. Possible physical consequences of the suggested models are listed and the possibility of their experimental finding is discussed. The functioning of the biomembrane's ion channel is qualitatively described on the basis of the suggested ferroactive models. The main directions and prospects for development of the ferroactive approach to the theory of biological membranes and their structures are indicated.

[Existence of boundary lipids in reconstituted hydrophobic protein-lipid system]. / Sushchestvovanie pogranichnykh lipidov v rekonstruirovannow sisteme gidrofobnyi belok--lipid.

A fraction of hydrophobic proteins (9, 14, 18 kD) soluble in a chloroform-methanol mixture (2:1) has been isolated from Micrococcus lysodeikticus bacterial membranes. The proteins obtained were introduced into proteoliposomes at a protein/lipid weight ratio ranging from 0.1 to 0.25 in combination with the fluorescent probe pyrene or the spin probe 2-(14-carboxytetradecyl)-2-ethyl-4.4-dimethyl-3-oxasolidinyloxyl. The excimertization of pyrene upon direct excitation of its molecules (gamma excit.=338nm) and under conditions of energy transfer from the excited protein chromophores to pyrene (gamma excit.=286nm) and the spin-spin exchange between the spin probe molecules was investigated. The experimental results suggest that the hydrophobic protein molecules are surrounded by a structurally heterogenous lipid area containing up to 3.3 mg of lipid per l mg of protein. The maximal expression of structural heterogeneity was observed at the minimal content of protein in the proteoliposomes. Treatment with the membranotropic antibiotic gramicidin S resulted in disappearance of lateral heterogeneity of lipids in the constituted system and in lipid aggregation in bacterial membranes. It is assumed that the aggregability of membrane proteins depends on the structural rearrangement of some part of lipid bilayer around them.

[Isolation of DNA-membrane complexes from Bacillus subtilis]. / Uydelenie i sostav DNK-membrannykh kompleksov iz Bacillus subtilis.

The DNA-membrane complexes were isolated from the membranes of Bacillus subtilis, disrupted by ultrasonication and subjected to chromatography on Sepharose 4B. The membrane-bound fraction of native 3H-DNA was found only in the freshly isolated material. The phospholipid and protein composition of this fraction was determined. It was assumed that the newly synthesized membrane-associated 3H-DNA from Bacillus subtilis is bound to the lipoprotein complexes enriched with phospholipids.

[Functional role of aminoacyl phosphatidylglycerols in the lamellar system of chloroplasts]. / K izucheniiu funktsional'noi roli aminoatsilfosfatidilgltserinov v lamelliarnoi sisteme khloroplastov.

The role of 14C-aminoacyl-tRNAs in the formation of aminoacyl phosphatidyl glycerols in isolated chloroplasts of haricot bean leaves was studied. The formation of 14C-aminoacyl-tRNAs was more intensive in the case when 14C-aminoacyl phosphatidyl glycerols were the source of amino acids. On incubation of lamellae with 14C-aminoacyl phosphatidyl glycerols, 14C-amino acids proved to be incorporated intensively in protein of the lamellae. Membrane-bound chloroplast ribosome-like particles were observed on the outermost thylakoid membranes of the grana stacks as well as on the stroma thylakoids. It is concluded that aminoacyl phosphatidyl glycerols play an important role in lateral transport of amino acids within the chloroplasts lamellar system.

[The aminoacyl phosphatidyl glycerols of the plastid membrane system in the process of chloroplast biogenesis]. / Aminoatsilfosfatidilglitseriny membrannoi sistemby plastid pri biogeneze khloroplastov.

The composition of aminoacyl phosphatidyl glycerols in maize plastids at different stages of chloroplast differentiation has been studied. In the course of incubation of 14C amino acids or 14CO2 with maize and bean seedlings in vivo the 14C amino acids were incorporated preferably into the acid phospholipid fraction, forming O-esters of amino acids with phosphatidylglycerols. The rate of lipoamino acid compounds formation increased with the chloroplast differentiation and reached its maximum in the seedlings containing chloroplasts with a developed lamellar system. Changes in the amino acid composition of 14C aminoacyl phosphatidyl glycerols were observed at all stages of chloroplast ultrastructure development.

[Incorporation of 32P into membrane phospholipids during chloroplast biogenesis]. / Vkliuchenie P32 fosfolipidy membran pri biogeneze khloroplastov.

The composition of membrane phospholipids during chloroplast biogenesis was studied. The maximal level of phosphatidic acid was observed in the membrane fraction of proplastids. Phosphatidylglycerol was found to be the most abundant phospholipid component of grana thylakoids. The evidence from the in vivo experiments indicates that phosphatidic acid and phosphatidylglycerol incorporate the 32P label at a high rate at all stages of the chloroplast biogenesis. It is concluded that plastids are the site of the phosphatidylglycerol biosynthesis in the plant cell.

[The nature of peptides firmly associated with the protein part of membrane lipoproteins in chloroplast biogenesis]. / O prirode peptidov, prochno assotsiirovannykh a belkovoi chast'iu lipoproteidov membran pri biogeneze khloroplastov.

The biosynthesis of membrane lipoproteins in maize plastids at different stages of differentiation of the chloroplast lamellar system was studied. Prolamellar and lamellar system preparations were isolated from maize plastids and been or kidney bean chloroplasts, disintegrated by osmotic shock. Chloroplast lipoproteins are membraneous proteins of chloroplast lamellar systems. Peptids, lipids and carbohydrates "firmly" associated with the protein part of membrane lipoproteins were found at all stages of chloroplast ultrastructure formation. Incorporation of C14 amino acid increased with the time of incubation in "firmly" lipoprotein--bound peptids and in the polypeptid part of lipoproteins. Puromycin but not actidion D inhibited amino acid incorporation into the peptids and polypeptids of chloroplasts in vitro experiments. The maximal level of C14-peptids "firmly" associated with the protein part of membrane lipoproteins was observed in the membrane fraction of proplastids in vivo. It is pointed out that biogenesis process leads to the decrease of content of "firmly" lipoprotein--bound peptids in the chloroplast lamellae fraction.

[The glycolipoproteins of plastid membrane system in process of chloroplast biogenesis]. / Glikolipoproteidy membrannoi sistemy plastid pri biogeneze khloroplastov

The glycolipoprotein fractions were isolated from the prolamellar and lamellar system of plastids. The glycolipoproteins were studied on the formic acid--urea polyacrylamide gel. The glycolipoproteins from the chloroplast lamellar system of bean leaves have the molecular weight 38 000, 93 000 and above 160 000. The amino acid composition of glycolipoproteins and its biosynthesis were studied in vivo and in vitro experiments. The glycolipoproteins were the first membrane proteins which were formed by chloroplasts in vitro. It is concluded that 70S ribosomes are involved in the glycolipoproteins biosynthesis.

[Study of amino acid composition and biosynthesis of protein components of the membrane system of corn plastids during biogenesis of chloroplasts]. / Izuchenie aminokislotnogo sostava i biosinteza belkovykh komponentov membrannoi sistemy plastid pri biogeneze khloroplastov

The biosynthesis of membrane proteins in maize plastids at different stages of differentiation of the chloroplast lamellar system was studied. Prolamellar and lamellar system preparations were isolated from maize plastids, disintegrated by osmotic shock under hypotonic conditions. Changes in the amino acid composition of 14C membrane proteins were observed at all stages of chloroplast ultrastructure formation. The maximal level of the apolar amino acids was observed in the membrane fraction of chloroplasts. Washed membranes from maize proplastids and chloroplasts can be resolved into at least 14 protein bands on formic acid--urea polyacrylamide gel. It is pointed out that biogenesis process leads to the increase of lipophylic protein content in the chloroplast lamellae fraction.