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PubMed2004

[Surface architectonics and cytoskeleton of erythrocytes and their modulation by adrenergic agents in patients with bronchial asthma].

AIM: To evaluate erythrocyte surface architectonics and cytoskeleton in bronchial asthma (BA) under modulation of adrenergic agents. MATERIAL AND METHODS: 24 healthy persons, 61 patients with bronchial asthma (BA) were examined. Of them, 28 patients had allergic BA (ABA) and 33 ones had nonallergic BA (NABA). Erythrocyte surface architectonics was studied by phase-contrast microscopy. Fixation in 0.5% solution of glutar aldehyde was used, preparation "squashed drop" was prepared. In all preparations adrenalin hydrochloride and obzidan were used in equimolar final concentrations of 4.10(-5) M. The mean morphologic index of transformation that reflects the shift to stomatocytosis or to echinocytosis was estimated to characterize integrally erythrocyte surface architectonics. The cytoskeleton was studied by the modified method of Chentosov's et al. The integral optic density was evaluated on imaging analyzer of Ista-Videotest company (St-Petersburg). RESULTS: In healthy persons there was a correlation between erythrocyte surface architectonics and cytoskeleton under modulation of adrenergic system. Adrenalin-induced decrease of cytoskeletal proteins correlated with the shift to stomatocytosis. In NABA erythrocyte surface architectonics was characterized by a pronounced shift to stomatocytosis. This shift was accompanied with the most pronounced decrease of cytoskeletal proteins and it reduced in remission. In ABA manifestation of the shift was minimal and did not depend on the phase of the disease. In ABA no changes in erythrocyte surface architectonics in the presence of adrenalin were revealed. In the same conditions a decrease in cytoskeletal proteins was found. CONCLUSION: A correlation was found between erythrocyte surface architectonics and cytoskeleton in healthy persons. In ABA, under modulation of adrenergic system by adrenalin dissociation was revealed between receptor and cytoskeletal mechanisms of cell form induction. This dissociation does not depend on the phase of the disease and is thought to be one of the important postreceptor disorders typical for this variant of the disease.

Adrenergic beta-Antagonists↗
PubMedDate not supplied

[Effect of L-carnitine on metabolic disorders in rats with experimental acyl-CoA dehydrogenase deficiency].

Effect of L-carnitine (LC) on the metabolism of organic acids and carnitine homeostasis was studied in rats with riboflavin deficiency producing unusual dicarboxylic acidurea and modeling multiple acyl-CoA dehydrogenase deficiency in humans. Riboflavin deficient (RFD) rats exhibited increased excretion of glutaric, ethylmalonic, and methylsuccinic acids, as well as isovaleryl-, butyryl-, isobutyryl-, 2-methyl-butyryl-, and hexanoylglycine, short-chain and medium-chain saturated, and unsaturated dicarboxylic organic acids (C6-C10). RFD rats also showed a decrease in the concentration of free LC in the blood plasma and in tissues, an increase in the level of isobutyryl- and isovalerylcarnitine in muscle tissue, and reduction in the level of acetyl- and propionylcarnitine in the blood plasma, kidney, and liver (all changes detected relative to animals in the control group). The introduction of LC to RFD rats normalized the LC homeostasis by increasing free LC concentration in the blood plasma and tissues, enhanced the acyl-LC excretion with urine and the level in tissues, and reduced the manifestations of organic acidurea.

Acyl-CoA Dehydrogenase↗
PubMedDate not supplied

[Activation of plasminogen in its immobilization].

Plasminogen is established to be able of transforming into the plasmin-like is compound when it is immobilized on aminoethyl cellulose by means of glutaric aldehyde. The plasminogen immobilized in such a way manifests a caseinolytic activity and is not activated additionally by streptokinase. The plasminogen immobilized on bromocyanogen-activated sepharose and cellulose, like soluble proenzyme, has no activity of plasmin and retains the ability of being activated with streptokinase in catalytic amounts.

Caseins↗
PubMedDate not supplied

Contribution of chitosan and its derivatives to cancer chemotherapy.

The conjugates of some kinds of anticancer agents with chitin and chitosan derivatives display good anticancer effects with a decrease in the adverse effects of the original drug due to a predominant distribution into the cancer and a gradual release of free drug from the conjugates. For instance, doxifluridine and 1-beta-D-arabinofuranosylcytosine (Ara-C) were conjugated with chitosan via glutaric spacer, and the conjugates of Ara-C with chitosan, in particular, showed a good antitumour effect against P388-bearing leukemia model mice. Glycol-chitosan (G-Chi) was distributed mainly in the systemic circulation and the kidney after i.v. administration into normal mice, and retained long in the kidney. The therapeutic effect of the conjugates of mitomycin C (MMC) with G-Chi was not necessarily improved in comparison with that of the free drug, but toxic side-effects appeared to decrease with the conjugates. The conjugates of MMC with 6-O-carboxymethyl-chitin showed almost complete suppression of tumour growth at 10 mg eq. MMC/kg, though a lethal adverse effect was also observed. The conjugates of MMC with N-succinyl-chitosan showed good antitumour activities against various tumour models due to their predominant distribution into the tumour tissue and sustained-release characteristics, irrespective of water-insoluble and -soluble formulations. It is believed that the chitin and chitosan derivatives discussed in this review are good candidates for a polymeric drug carrier in cancer chemotherapy.

Animals↗
PubMed2003

Tandem mass spectrometric analysis for disorders in amino, organic and fatty acid metabolism: two year experience in South Korea.

Seoul Clinical Laboratories began screening newborns and high risk group blood spots with tandem mass spectrometry (MS/MS) in April 2001. The goal was to determine approximate prevalence of metabolic disorders and optimization of decision criteria for estimation of preventive effect with early diagnosis. Approximately 44,300 neonates and children were screened and the estimated prevalence (newborn/high risk group), sensitivity, specificity and recall rate amounted to 1:2000 / 1:1250, 94.1 %, 99.7 %, and 0.04 %, respectively. Confirmed 35 multiple metabolic disorders (newborn/high risk) were as follows; 16 amino acid disorders [classical PKU(3/4), BH4 deficient-hyperphenylalaninemia(0/1), Citrullinemia(2/0), Homocystinuria(0/2), Hypermethioninemia(0/1), Tyrosinemia(1/0)], OTC deficiency (0/1), MSUD (2/0), 10 organic acidurias [Propionic aciduria(2/1), Methylmalonic aciduria(0/1), Isovaleric aciduria(2/1), 3-methylcrotonylglycineuria(1/0), Glutaric aciduria type 1(2/0)], 9 fatty acid oxidation disorders [LCHAD def. (2/2), Mitochondrial TFP def.(0/1), VLCAD def.(1/0), LC3KT def.(0/1), SCAD def (1/0), MADD def (0/1). The relatively normal development of 15 patients with metabolic disorders among newborns (except for the expired) demonstrates the usefulness of newborn screening by MS/MS for early diagnosis and medical intervention. However, close coordination between the MS/MS screening laboratory and the metabolic clinic/biochemical geneticists is needed to determine proper decision of screening parameters, confirmation diagnosis, follow-up scheme and additional tests.

Amino Acids↗
PubMed2005

[Influence of tobacco smoking on lipase activity in patients with pancreatitis].

The aim of this study is to prove the influence of tobacco smoking on lipase activity in the blood of smoking and non-smoking health persons and in smoking and non-smoking patients with diagnosed acute (AP), chronic exaggerated (CEP) and chronic pancreatitis (CP). The blood has been collected from 28 healthy persons and 55 patients with AP, CEP and CP. The enzyme activity has been determined using the colorimetric method with substrate 1,2-odilauryl-rac-glycero-3-glutaric acid -(6-methylresorufin) ester. The exposures to tobacco smoke have been examined on the basic of concentration of cotinine in the serum of patients. The highest lipase activity has been found in smoking patients with CEP. It has been noted that the serum lipase activity is significantly higher in smoking and healthy persons (p<0,05) then in non-smoking and healthy patients. However no significant differences have been found between the lipase activity in smoking patients with CP and non-smoking patients with CP. Smoking patients with AP and CEP have been found to have a significantly increased enzyme activity (p>0.01; p>0.05 respectively) when compared to non-smoking patients. Results of examination indicate that tobacco smoking has a significant influence on exocrine function of pancreas.

Acute Disease↗
PubMed1991

NMR observation of exchangeable protons of pyridoxal phosphate and histidine residues in cytosolic aspartate aminotransferase.

Observation of the 93-kDa cytosolic aspartate aminotransferase by 500-MHz 1H NMR spectroscopy in H2O has revealed a series of resonances in the 10-18 ppm range arising from exchangeable protons. One of these (peak A) has been assigned to the proton bound to the ring nitrogen of the coenzyme pyridoxal 5'-phosphate. A second (peak B) is assigned to H143 which participates in a chain of hydrogen bonds that includes also the coenzyme-bound proton. There is a mutual nuclear Overhauser effect between these two resonances. Peaks A and B respond to changes in pH and to interaction of the enzyme with coenzyme derivatives and inhibitors. Peak A moves from 15.4 to 17.4 ppm as the pH is lowered, while peak B moves in the opposite direction from 14.7 to 13.7 ppm, both with an apparent pKa of 6.15. This pKa is associated with deprotonation of the imine nitrogen at the Schiff base linkage of the coenzyme with K258 of the enzyme. In spectra of enzyme containing pyridoxamine 5'-phosphate, peak A is observed at 16.5 ppm and peak B is at 13.9 ppm over a broad pH range. Peaks A and B are found at 17.8 and 14.0 ppm, respectively, for the enzyme complex with glutarate. When alpha-methylaspartate is added to the enzyme several new resonances appear in the spectrum, which are attributed to formation of the external aldimine. The position of peak A in spectra of various forms of the enzyme is interpreted to reflect the electronic distribution in the coenzyme ring. Several other peaks in this region of the spectrum also are sensitive to changes in pH or the addition of inhibitors. Some possible assignments of these resonances are discussed.

Aspartate Aminotransferases↗
PubMed2006

[Ultrastructural study of the pulmonary veins in pigs].

OBJECTIVE: To explore the possible relationship between the ultrastructural characteristics of pulmonary veins and the pathogenesis of atrial fibrillation originating from pulmonary veins. METHODS: The pulmonary veins from domestic pigs were serially sectioned (2 mm) transversely along the vessels. The odd number sections were fixed in 10% phosphate buffered formalin solution and the even number sections were fixed in 3% Glutaral for further electron microscopy observations. RESULTS: Two cell types were found in the pulmonary veins of pigs. One cell type was the P-like cells that had an empty-appearing cytoplasm containing only sparse myofibrils and small mitochondria, both of which were randomly distributed. Another cell type was slender transitional cells with plenty of longitudinally displayed myofibrils. CONCLUSION: P-like cells in the pig pulmonary veins were found using electron microscopy in this study and ectopic beats from P-like cells in the myocardial sleeves in the pulmonary veins might be responsible for atrial fibrillation originating from pulmonary veins.

Animals↗
PubMed2006

A report of two families with sarcosinaemia in Hong Kong and revisiting the pathogenetic potential of hypersarcosinaemia.

INTRODUCTION: Sarcosinaemia is a rare metabolic disorder which has not been reported in Asia. CLINICAL PICTURE: The urine samples of 2 patients were screened as a routine metabolic screening offered for patients with mental retardation in our hospital. We used gas chromatography-mass spectrometry (GC-MS) which is capable of detecting abnormal pattern in amino acids and organic acids. Plasma sarcosine level was further quantified by GC-MS. The same methods were used in the investigations of asymptomatic family members. Urine examination by GC-MS revealed excessive amount of sarcosine in urine (normally undetectable) and their plasma sarcosine levels were raised. The 2 differential diagnoses of presence of sarcosine in urine--glutaric aciduria type II and folate deficiency--were ruled out by the absence of abnormal organic acids in the initial urine screen and by normal serum folate level respectively. Screening of the 2 families identified excessive sarcosine in urine in 2 siblings, one from each family. However, these 2 siblings of indexed patients thus identified have no neurological or developmental problem. CONCLUSION: Our finding was consistent with the notion that sarcosinaemia is a benign condition picked up coincidentally during screening for mental retardation.

Amino Acid Metabolism, Inborn Errors↗
PubMed1990

[The ultrastructural changes occurring in the endometrium of women who use intrauterine pessaries].

The authors studied ultrastructural changes in the endometrium of 25 women, using intrauterine contraceptive devices, preventing conception, during various time limits. The material of the endometrial biopsy was fixed with glutar and osmium in a phosphate buffer and embedded in durcopan. Section were prepared by a microtome and observed under electron microscope of Hitachi type. Comparing the parameters duration of usage of loops and character of changes in the endometrium an impression was obtained that the usage of contraceptive intrauterine devices for one-year period did not induced changes of pathological character. Functional changes in the endometrium occurred after a two-year stay of the loop in the uterus as there were inflammatory changes of spot-like character in two of the sections. Local areas of inflammation of nonspecific character, manifested under the form of lymphocytes and leucocyte accumulation, were found in women with long usage of contraceptive intrauterine devices. The authors advise to remove the loop 2.5-3 years after its application and to wait for the complete regeneration of the endometrium.

Endometritis↗
PubMed1991

[The pavement lining of the vessels in the earthworm (Lumbricus terrestris)].

Effect of some preparatory methods to make ready earthworm tissues for electron microscopical investigation has been studied in order to reveal structural components of the vascular wall. The method for an immediate fixation of the earthworm tissues in liquid nitrogen with a subsequent additional fixation in cooled 2.5% glutar aldehyde, applied in combination with OsO4-thiocarbohydroside-OsO4 method is optimal. This method of samples preparation gives a reliable information concerning existence of an internal--endothelial--lining in the earthworm vessels. A conclusion is made on a qualitatively new level of cell differentiation of the internal vascular lining in the earthworm, which is called phase of blast transformation of endotheliocytes.

Animals↗
PubMed2006

[The immune enhancement of the C3 to the Escherichia coli vaccine].

C3b was separated and purified from the SPF chicken serum. It was linked with E. coli antigen by the glutaral. 11 days aged SPF chicken were immunized by the complex antigen and the chickens of control group were immunised by the FCA- E. coli antigen . They were boosted at the age of 18 day. The immune response was monitored by an enzyme-linked immunosorbent assay(ELISA) for anti-E. coli anitibody. The ELISA results indicated that during the early several weeks, IgG titers elicited by FCA (FCA-E. coli) were higher than those elicited by C3 (C3b-E. coli), but decreased rapidly after a peak around the end of 4th week from being immunized. Chickens immunized with C3b always gave increased response, and the IgG titers were equal to that of FCA at the end of 5th week from being immunized and then higher and higher than that of FCA. Thus the adjuvant effect of C3b is different from that of FCA, it could induce production of memory cell and make the antigens stimulate immune cells consistently and stably.

Adjuvants, Immunologic↗
PubMed1975

Saccharopine dehydrogenase. Substrate inhibition studies.

In the direction of reductive condensation of alpha-ketoglutarate and lysine, saccharopine dehydrogenase (N6-(glutar-2-yl)-L-lysine:NAD oxidoreductase (lysine-forming) is inhibited by high concentrations of alpha-ketoglutarate and lysine, but not by NADH. NAD+ and saccharopine show no substrate inhibition in the reverse direction. Substrate inhibition by alpha-ketoglutarate and lysine is linear uncompetitive versus NADH. However, when the inhibition is examined with alpha-ketoglutarate or lysine as the variable substrate, the double reciprocal plots show a family of curved lines concave up. The curvature is more pronounced with increasing concentrations of the inhibitory substrate, suggesting an interaction of variable substrate with the enzyme form carrying the inhibitory substrate. These inhibition patterns, the lack of interaction of structural analogs of lysine such as ornithine and norleucine with the E-NAD+ complex (Fujioka M., and Nakatani, Y. (1972) Eur. J. Biochem. 25, 301-307), the identity of values of inhibition constants of alpha-ketoglutarate and lysine obtained with either one as the substrate inhibitor, and the substrate inhibition data in the presence of a reaction product, NAD+, are consistent with the mechanism that substrate inhibition results from the formation of a dead-end E-NAD+-alpha-ketoglutarate complex followed by the addition of lysine to this abortive complex.

Ketoglutaric Acids↗
PubMed1991

Differentiation of Salmonella senftenberg into biogroups.

Ninety-six strains of Salmonella senftenberg, isolated between 1984 and 1986 from different parts of England and Wales, were tested for their biochemical reactions and biotyped according to the method of Duguid and others (1975). Nine biogroups were identified on the basis of their metabolism of L-tartrate, D-tartrate, Bitter's xylose and Stern's glycerol. In addition, fumaric, oxalic, succinic, glutaric, malonic, maleic, L-malic, L-aspartic, lactic and formic acids were used but did not increase the discrimination. Three biogroups (7, 2 and 5) accounted for 79 per cent of the cultures examined.

Bacterial Typing Techniques↗
PubMed1975

[Role of adenine mono- and dinucleotides in ammonia formation in brain tissue].

The investigations carried out have shown that not only AMP but ADP also undergoes direct deamination in both soluble and mitochondrial fractions of rat brain tissue. Deamination of AMP is stimulated by the addition of ATP and the activity of one of the isoenzymes of AMP-aminohydrolase is markedly enhanced by both yeast and brain hexokinase. Activation by hexokinase is mainly due to its SH groups, through which hexokinase reacts with AMP-aminohydrolase, forming, probably, a protein-protein complex in which AMP aminohydrolase activity is considerably increased. Hexokinase does not affect the deamination of ADP and NAD. Further experiments are needed to find out whether the activation of AMP-aminohydrolase is accomplished by hexokinase itself or by an other protein contaminating it. Deamination of NAD, in contrast to AMP and ADP, takes place only in mitochondria and does not occur in the soluble fraction. In mitochondria besides deamination, AMP and ADP undergo intensive dephosphorylation, while the deamination of NAD is not accompanied by an increase of phosphate, i. e. mitochondria lack enzymes which breakdown NAD to mono nucleotides. Our data indicate that the formation of deamino -NAD from NAD and reamination of deamino-NAD by aspartate to NAD by the formation of intermediary NAD-succinate is of greater importance. The formation of the latter and that of deamino-NAD from NAD as well as the presence of preformed deamino-NAD in mitochondria have been demonstrated by Movsessian. The occurrence of these processes in mitochondria and their role in the formation of ammonia from amino acids is of importance in as much as oxaloacetate formation and its conversion to aspartate, which is necessary for the reamination of deamino-NAD, are localized in mitochondria. The main source of the amino nitrogen of aspartate is known to be glutamate, which incorporates the amino nitrogen of most amino acids. alpha-Keto-glutarate, which is necessary for the synthesis of glutamate, is also formed in mitochondria are the most favourable site for the formation of ammonia from amino acids with the participation of pyridine nucleotides. Of the purine mono and dinucleotides studied deamino-NAD is most effective in the formation of ammonia from amino acids in mitochondria since in contrast to purine mono nucleotides, deamino-NAD and NAD are not dephosphorylated in mitochondria. According to some authors the reamination of IMP by aspartate is of importance in the formation of ammonia from amino acids in brain tissue. In our studies, however, IMP was not effective in the formation of ammonia from aspartate in mitochondrial fractions. IDP was found to be more effective. IMP and IDP may probably participate in the formation of ammonia in the soluble fraction, where nucleotidase activity is considerably low.

AMP Deaminase↗
PubMed1991

The role of His143 in the catalytic mechanism of Escherichia coli aspartate aminotransferase.

In aspartate aminotransferase (AspAT), His143 is located within a hydrogen-bonding distance to Asp222 that forms a strong ion pair with the ring nitrogen of the coenzyme, pyridoxal 5'-phosphate (PLP) or pyridoxamine 5'-phosphate (PMP). His143 of Escherichia coli AspAT was replaced by Ala or Asn. The mutant enzyme H143A showed a slight increase in the maximum velocity of the overall transamination reaction between aspartate and 2-oxoglutarate, while H143N AspAT showed a decrease to 60% in the maximum rate of the overall reactions in both directions. In all of the half-transamination reactions with four substrates, aspartate, glutamate, oxalacetate, and 2-oxoglutarate, the catalytic competence as defined by kmax/Kd decreased by 3-18-fold upon replacing His143 by either Ala or Asn. The extent of the decrease varied from one substrate to another; it was largely contributed to by the decrease in affinities for all substrates. The equilibrium constants, [PMP-form] [keto acid]/[( PLP-form] [amino acid]), decreased by over 10-fold upon the mutations at position 143. Both H143A and H143N AspATs exhibited a considerably decreased affinity for 2-methylaspartate, an external-aldimine-forming substrate analogue, yet without appreciable alteration in the affinity for succinate and glutarate, which are non-aldimine-forming analogues. All these findings suggest that, although His143 is not essential for catalysis, it might assist the formation of enzyme-substrate complex.

Aspartate Aminotransferases↗
PubMed1977

[Immunogenicity and thymus-dependence of the polymerized alpha-toxoid of Clostridium perfringens].

Polymeric alpha-toxoid with a molecular weight of 450 000--600 000 was obtained by condensation of alpha-toxoid of Cl. perfringens, type A, with glutaric aldehyde. Experiments on guinea pigs showed that in the adsorbed preparations the immunogenic properties of both monomeric and polymeric alpha-toxoids are practically identical. The primary immune response after the immunization with nonadsorbed antigens was 3 times greater than that induced by the monomer. Polymerization of alpha-toxoid failed to change its thymus dependency.

Animals↗
PubMed1990

Specificity of lipoamide dehydrogenase for alpha-keto acid dehydrogenase complexes and the glycine cleavage system.

Rat liver lipoamide dehydrogenase (LipDH) was separated into three types on DE-32 column chromatography, but no difference was observed among them in either immunological reactivity or enzymatic properties. A reconstitution experiment of branched-chain alpha-keto acid dehydrogenase complex (BCKADH) revealed that the most anionic type of LipDH was the most effective for the enzyme complex while the three types of LipDH were the same in the affinity for BCKADH subcomplex. All three types of LipDH were equally effective in reconstituting pyruvate dehydrogenase complex, alpha-ketoglutarate dehydrogenase complex and the glycine cleavage system. However, either pyruvate dehydrogenase or alpha-keto-glutarate dehydrogenase complex appeared to involve a certain LipDH in vivo which was firmly integrated into and hardly dissociable from the complex. A broad specificity of LipDH was observed for the glycine cleavage system. When BCKADH reconstitution experiments were carried out with both LipDHs from various sources and purified rat liver BCKADH subcomplex, the effectiveness of animal LipDHs was proportional to the extent of their immunological reactivity to the anti-rat LipDH antibody. However, BCKADH activity was also restored by a certain bacterial LipDH which had no cross-reactivity with the antibody, and LipDHs from some bacterial species, which reacted well with the antibody, showed no effect for the reconstitution of BCKADH. Thus, the determinant(s) of LipDH for the integration into alpha-keto acid dehydrogenase complexes including BCKADH can be its tertiary and/or quarternary structure rather than its primary and secondary structures.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗