Peripheral brain-derived neurotrophic factor is related to cardiovascular risk factors in active and inactive elderly men.

Regular exercise plays an important preventive and therapeutic role in heart and vascular diseases, and beneficially affects brain function. In blood, the effects of exercise appear to be very complex and could include protection of vascular endothelial cells via neurotrophic factors and decreased oxidative stress. The purpose of this study was to identify the age-related changes in peripheral brain-derived neurotrophic factor (BDNF) and its relationship to oxidative damage and conventional cardiovascular disease (CVD) biomarkers, such as atherogenic index, C-reactive protein (hsCRP) and oxidized LDL (oxLDL), in active and inactive men. Seventeen elderly males (61-80 years) and 17 young males (20-24 years) participated in this study. According to the 6-min Åstrand-Rhyming bike test, the subjects were classified into active and inactive groups. The young and elderly active men had a significantly better lipoprotein profile and antioxidant status, as well as reduced oxidative damage and inflammatory state. The active young and elderly men had significantly higher plasma BDNF levels compared to their inactive peers. BDNF was correlated with VO2max (r=0.765, P<0.001). In addition, we observed a significant inverse correlation of BDNF with atherogenic index (TC/HDL), hsCRP and oxLDL. The findings demonstrate that a high level of cardiorespiratory fitness reflected in VO2max was associated with a higher level of circulating BDNF, which in turn was related to common CVD risk factors and oxidative damage markers in young and elderly men.

Peripheral brain-derived neurotrophic factor is related to cardiovascular risk factors in active and inactive elderly men

Regular exercise plays an important preventive and therapeutic role in heart and vascular diseases, and beneficially affects brain function. In blood, the effects of exercise appear to be very complex and could include protection of vascular endothelial cells via neurotrophic factors and decreased oxidative stress. The purpose of this study was to identify the age-related changes in peripheral brain-derived neurotrophic factor (BDNF) and its relationship to oxidative damage and conventional cardiovascular disease (CVD) biomarkers, such as atherogenic index, C-reactive protein (hsCRP) and oxidized LDL (oxLDL), in active and inactive men. Seventeen elderly males (61-80 years) and 17 young males (20-24 years) participated in this study. According to the 6-min Åstrand-Rhyming bike test, the subjects were classified into active and inactive groups. The young and elderly active men had a significantly better lipoprotein profile and antioxidant status, as well as reduced oxidative damage and inflammatory state. The active young and elderly men had significantly higher plasma BDNF levels compared to their inactive peers. BDNF was correlated with VO2max (r=0.765, P<0.001). In addition, we observed a significant inverse correlation of BDNF with atherogenic index (TC/HDL), hsCRP and oxLDL. The findings demonstrate that a high level of cardiorespiratory fitness reflected in VO2max was associated with a higher level of circulating BDNF, which in turn was related to common CVD risk factors and oxidative damage markers in young and elderly men.

Influence of l-carnosine on pro-antioxidant status in elite kayakers and canoeists.

Carnosine is a dipeptide formed from the amino acids ß-alanine and histidine and found in large amounts in the brain and muscle, especially fast twitch muscle. Carnosine has an antioxidant role and accounts for about 10% of the muscle's ability to buffer the H+ ions produced by high intensity exercise. Due to the interesting role of carnosine, the aim of the study was observe the effects of carnosine intake on pro-antioxidant status in highly trained athletes exposed to intense exercise.Fourteen male athletes from the Polish national kayak and canoe teams participated in placebo-controlled and cross-over study. The athletes were supplemented with 4 g/d carnosine for 14 days. Blood samples were collected before and 30 min, 24 h and 48 h after 2000 m exercise trial. In blood, hydrogen peroxide (H2O2), nitric oxide (NO), markers of RO/NS activity 8-isoprostanes and 3-nitrotyrosine, total (GSHt) and oxidised glutathione (GSSG), antioxidant status (APO) and superoxide dismutase (SOD) were determined. There were not observed statistically significant differences in exercise-induced changes in H2O2 and NO concentrations and SOD activity after carnosine intake. However, carnosine prevented an increase in 8-isoprostanes, 3-nitrotyrosine and GSSG concentrations as well as elevated redox status (GSHt-2GSSG)/GSSG at post-exercise period.Although, oral supplementation with 4 g carnosine did not affect RO/NS generation, it significantly attenuated exercise-induced glutathione loss, reduced oxidation/nitration markers concentration and SOD activity. These results suggest that carnosine could provide antioxidative protection for highly trained athletes.

Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade.

The x-ray crystal structure of recombinant trichodiene synthase from Fusarium sporotrichioides has been determined to 2.5-A resolution, both unliganded and complexed with inorganic pyrophosphate. This reaction product coordinates to three Mg(2+) ions near the mouth of the active site cleft. A comparison of the liganded and unliganded structures reveals a ligand-induced conformational change that closes the mouth of the active site cleft. Binding of the substrate farnesyl diphosphate similarly may trigger this conformational change, which would facilitate catalysis by protecting reactive carbocationic intermediates in the cyclization cascade. Trichodiene synthase also shares significant structural similarity with other sesquiterpene synthases despite a lack of significant sequence identity. This similarity indicates divergence from a common ancestor early in the evolution of terpene biosynthesis.

Crystal structure determination of aristolochene synthase from the blue cheese mold, Penicillium roqueforti.

The 2.5-A resolution crystal structure of recombinant aristolochene synthase from the blue cheese mold, Penicillium roqueforti, is the first of a fungal terpenoid cyclase. The structure of the enzyme reveals active site features that participate in the cyclization of the universal sesquiterpene cyclase substrate, farnesyl diphosphate, to form the bicyclic hydrocarbon aristolochene. Metal-triggered carbocation formation initiates the cyclization cascade, which proceeds through multiple complex intermediates to yield one exclusive structural and stereochemical isomer of aristolochene. Structural homology of this fungal cyclase with plant and bacterial terpenoid cyclases, despite minimal amino acid sequence identity, suggests divergence from a common, primordial ancestor in the evolution of terpene biosynthesis.

Fate of dietary sucrose and neosynthesis of amino acids in the pea aphid, acyrthosiphon pisum, reared on different diets

The fate of sucrose, the major nutrient of an aphid's natural food, was explored by radiolabeling in the pea aphid Acyrthosiphon pisum. To investigate the influence of nitrogen quality of food on amino acid neosynthesis, pea aphids were reared on two artificial diets differing in their amino acid composition. The first (diet A) had an equilibrated amino acid balance, similar to that derived from analysis of aphid carcass, and the other (diet B) had an unbalanced amino acid composition similar to that of legume phloem sap. Aphids grown on either diet expired the same quantity of sucrose carbon as CO(2), amounting to 25-30 % of the ingested sucrose catabolized in oxidation pathways. On diet A, the aphids excreted through honeydew about twice as much sucrose carbon as on diet B (amounting to 12.6 % of the ingested sucrose for diet A and 8.4 % for diet B), while amounts of sucrose carbons incorporated into exuviae were almost identical (1.9 % of the ingested sucrose on diet A and 2.7 % on diet B). There was also no difference in the amounts of sucrose carbon incorporated into the aphid tissues, which represented close to 50 % of the ingested sucrose. Sucrose carbons in the aphid tissues were mainly incorporated into lipids and the quantities involved were the same in aphids reared on either diet. On diet B, we observed neosynthesis of all protein amino acids from sucrose carbons and, for the first time in an aphid, we directly demonstrated the synthesis of the essential amino acids leucine, valine and phenylalanine. Amino acid neosynthesis from sucrose was significantly higher on diet B (11.5 % of ingested sucrose carbons) than on diet A (5.4 %). On diet A, neosynthesis of most of the amino acids was significantly diminished, and synthesis of two of them (histidine and arginine) was completely suppressed. The origin of amino acids egested through honeydew was determined from the specific activity of the free amino acid pool in the aphid. Aphids are able to adjust to variation in dietary amino acids by independent egestion of each amino acid. While more than 80 % of excreted nitrogen was from food amino acids, different amino acids were excreted in honeydew of aphids reared on the two diets. The conversion yields of dietary sucrose into aphid amino acids determined in this study were combined with those obtained previously by studying the fate of amino acids in pea aphids reared on diet A. The origin of all the amino acid carbons in aphid tissues was thus computed, and the metabolic abilities of aphid are discussed from an adaptive point of view, with respect to their symbiotic status.

Chemical rescue by guanidine derivatives of an arginine-substituted site-directed mutant of Escherichia coli ornithine transcarbamylase.

Escherichia coli ornithine transcarbamylase (OTCase) catalyzes the production of L-citrulline and phosphate from carbamyl phosphate and L-ornithine in L-arginine biosynthesis. We show that exogenous guanidines can restore activity to (chemically rescue) a catalytically-impaired site-directed mutant OTCase, R57G, in which glycine replaces an an active site arginine. The best rescue agent is guanidine hydrochloride, which enhances the rate of the mutant 2000-fold. The turnover number for the guanidine-rescued R57G mutant is 10% that of wild-type. The addition of guanidine to the R57G mutant has little effect on KMCP values, and the rescue effect is therefore attributed principally to an increase in kcat. Other compounds were screened as potential rescue agents, but rate enhancement is highly selective for guanidines. Not all guanidines show large increases in kcat. For a comparative series that includes guanidine and alkylguanidines, substituent size is inversely related to kcat. Brønsted analysis of guanidines with varying pKa values indicates that a partial positive charge is implicated in rescue, consistent with the proposed role of arginine 57 in catalysis. In UV difference and 31P-NMR spectra, carbamyl phosphate-induced effects associated with wild-type OTCase are observed in the R57G mutant only in the presence of guanidine. The kinetic mechanism of the mutant is random in the presence or absence of guanidine, in contrast to the sequential ordered mechanism of the wild-type enzyme. Thus, chemical rescue of R57G by guanidine hydrochloride restores many but not all wild-type properties to the mutant enzyme.

Transient changes in erythrocyte membrane permeability are induced by sublytic amounts of the complement membrane attack complex (C5b-9).

We have previously shown that sublytic heterologous complement induces large but transient increases in erythrocyte membrane permeability. We now report that when erythrocytes are bystanders in zymosan-activated autologous serum, they increase their Na+ permeability 10-fold, indicating that autologous complement can also induce transient membrane lesions. When we isolated the effect of the C5b-9 membrane attack complex of complement by using human C5b-9 assembled from purified components, we found there was minimal lysis but efficient Na+ uptake. Suspension of the sublytically damaged erythrocytes in K+ medium caused the cells to lyse, which is consistent with the cells recruiting a compensatory K+ efflux similar to that observed when human erythrocytes were exposed to heterologous complement. Sublytic C5b-9 exposure also became lytic when extracellular Ca2+ was limited and when the cells were exposed to charybdotoxin, an inhibitor of the Ca(2+)-activated K+ channel. This indicates that Ca2+ is required for the functional termination of the C5b-9 lesion. We also show that the membrane hyperpolarization resulting from activation of the Ca(2+)-dependent K+ efflux does not influence the termination of the C5b-9 lesion. Thus, the influx of Ca2+ through the complement lesion initiates at least two apparently independent adaptive responses: (1) a process that terminates the leak; and (2) a K+ efflux that has a volume regulatory function. Our data support the potential of the sublytic C5b-9 lesion to act as a physiologic mediator for autologous erythrocytes.