Content Of Coenzyme Research Articles

Potato glycoside alkaloids exhibit antifungal activity by regulating the tricarboxylic acid cycle pathway of Fusarium solani.

Fusarium solani is a pathogenic fungus that causes significant harm, leading to crop yield reduction, fruit quality reduction, postharvest decay, and other diseases. This study used potato glycoside alkaloids (PGA) as inhibitors to investigate their effects on the mitochondrial structure and tricarboxylic acid (TCA) cycle pathway of F. solani. The results showed that PGA could inhibit the colony growth of F. solani (54.49%), resulting in the disappearance of the mitochondrial membrane and the loss of contents. PGA significantly decreased the activities of aconitase (ACO), isocitrate dehydrogenase (IDH), α-ketoglutarate dehydrogenase (α-KGDH), succinate dehydrogenase (SDH), fumarase (FH), malate dehydrogenase (MDH), succinyl-CoA synthetase (SCS), and increased the activity of citrate synthase (CS) in F. solani. After PGA treatment, the contents of acetyl coenzyme A (CoA), citric acid (CA), malic acid (L-MA), and α-ketoglutaric acid (α-KG) in F. solani were significantly decreased. The contents of isocitric acid (ICA), succinyl coenzyme A (S-CoA), succinic acid (SA), fumaric acid (FA), and oxaloacetic acid (OA) were significantly increased. Transcriptomic analysis showed that PGA could significantly affect the expression levels of 19 genes related to TCA cycle in F. solani. RT-qPCR results showed that the expression levels of ACO, IDH, α-KGDH, and MDH-related genes were significantly down-regulated, and the expression levels of SDH and FH-related genes were significantly up-regulated, which was consistent with the results of transcriptomics. In summary, PGA can achieve antifungal effects by reducing the tricarboxylic acid cycle's flow and regulating key genes' expression levels. This study reveals the antifungal mechanism of PGA from the perspective of TCA cycle, and provides a theoretical basis for the development and application of PGA as a biopesticide.

Improvement in bacterial cellulose production by co-culturing Bacillus cereus and Komagataeibacter xylinus

Bacterial cellulose (BC) is a bio-produced nanostructure material widely used in biomedical, food, and paper-manufacturing industries. However, low production efficiency and high-cost have limited its industrial applications. This study aimed to examine the level of improvement in BC production by co-culturing Bacillus cereus and Komagataeibacter xylinus. The BC yield in corn stover enzymatic hydrolysate was found to be obviously enhanced from 1.2 to 4.4 g/L after the aforementioned co-culturing. The evidence indicated that acetoin (AC) and 2,3-butanediol (2,3-BD) produced by B. cereus were the key factors dominating BC increment. The mechanism underlying BC increment was that AC and 2,3-BD increased the specific activity of AC dehydrogenase and the contents of adenosine triphosphate (ATP) and acetyl coenzyme A (acetyl-CoA), thus promoting the growth and energy level of K. xylinus. Meanwhile, the immobilization of BC could also facilitate oxygen acquisition in B. cereus under static conditions. This study was novel in reporting that the co-culture could effectively enhance BC production from the lignocellulosic enzymatic hydrolysate.

Influence of raw pea (Pisum sativum) or blue lupin seeds (Lupinus angustifolius) on the level of selected bioactive substances in pork meat

Abstract The study objective was to evaluate the impact of different contributions of pea (Pisum sativum) cultivar Hubal and blue lupin (Lupinus angustifolius) cultivar Regent on the level of selected bioactive substances in pork meat. One hundred three cross-breed piglets: ♀ (Landrace × Yorkshire) × ♂ Duroc were used. Two experiments were performed, in which pea seeds (experiment I: E1 – 5.0% pea seeds; E2 – 10.0% pea seeds; E3 – 15.0% pea seeds; E4 – 17.5% pea seeds) and blue lupin seeds (experiment II; D1 – 5.0% blue lupin seeds; D2 – 10.0% blue lupin seeds; D3 – 15.0% blue lupin seeds D4 – 17.5% blue lupin seeds) were used instead of genetically modified soybean meal (SBM-GM). In each of the experiments 50 animals were divided into 5 groups (control – C, and four experimentals), placed in group pens, each for 10 individuals (sex ratio barrows : gilts – 1:1). The animals were weighed and tagged before the experiments. The initial body weight of the pigs at experiment I and II commencement was: 26.7 and 33.5 kg, and at the end of the experiments: 122.0 and 124.0, respectively. In the first experiment (progressive pea “seeds” contribution) the concentration of carnosine was shown to be higher in E4 than E3 and C by 47.3% and 94.2%, respectively. In comparison with group C, the Q10 coenzyme content in groups E1, E2, E3 and E4 was lower by 40.9%, 56.8%, 40.9% and 65.9% respectively. In the experiment II (progressive lupin “seeds” contribution) increased content of all of the investigated bioactive substances was recorded in groups D1-D3 vs C. Significant differences between groups C, D2, D4 for taurine (P≤0.05; P≤0.01) and creatine (P≤0.05) have been recorded. The content of bioactive substances in the longissimus lumborum muscle was significantly influenced by legumes, which increased the level of bioactive components of protein fraction. Therefore, it can be concluded that pea (Pisum sativum) cultivar Hubal and blue lupin (Lupinus angustifolius) cultivar Regent are an alternative to SBM-GM, increasing the nutritional value of pork meat.

A small heat shock protein, GmHSP17.9, from nodule confers symbiotic nitrogen fixation and seed yield in soybean

SummaryLegume–rhizobia symbiosis enables biological nitrogen fixation to improve crop production for sustainable agriculture. Small heat shock proteins (sHSPs) are involved in multiple environmental stresses and plant development processes. However, the role of sHSPs in nodule development in soybean remains largely unknown. In the present study, we identified a nodule‐localized sHSP, called GmHSP17.9, in soybean, which was markedly up‐regulated during nodule development. GmHSP17.9 was specifically expressed in the infected regions of the nodules. GmHSP17.9 overexpression and RNAi in transgenic composite plants and loss of function in CRISPR‐Cas9 gene‐editing mutant plants in soybean resulted in remarkable alterations in nodule number, nodule fresh weight, nitrogenase activity, contents of poly β‐hydroxybutyrate bodies (PHBs), ureide and total nitrogen content, which caused significant changes in plant growth and seed yield. GmHSP17.9 was also found to act as a chaperone for its interacting partner, GmNOD100, a sucrose synthase in soybean nodules which was also preferentially expressed in the infected zone of nodules, similar to GmHSP17.9. Functional analysis of GmNOD100 in composite transgenic plants revealed that GmNOD100 played an essential role in soybean nodulation. The hsp17.9 lines showed markedly more reduced sucrose synthase activity, lower contents of UDP‐glucose and acetyl coenzyme A (acetyl‐CoA), and decreased activity of succinic dehydrogenase (SDH) in the tricarboxylic acid (TCA) cycle in nodules due to the missing interaction with GmNOD100. Our findings reveal an important role and an unprecedented molecular mechanism of sHSPs in nodule development and nitrogen fixation in soybean.

Facial fabricated biocompatible homogeneous biocarriers involving biochar to enhance denitrification performance in an anoxic moving bed biofilm reactor

Biochar prepared from pineapple peel was facially combined with polyurethane sponges for the first time to form homogeneous biocompatible biocarriers, which can enhance denitrification performance in an anoxic MBBR. The experiments showed that a higher NO3–-N removal efficiency (96.24 ± 1.3%) and kinetic constant (0.26 h−1) were obtained in the MBBR employing these new biocarriers (B-MBBR), compared with a control MBBR with polyurethane sponges (C-MBBR). The attached and suspended biomass of the B-MBBR was increased by 47% and 26%, respectively. Biochar significantly enhanced the abundance of functional bacteria in terms of promoting biofilm (i.e., Leptonema), denitrifying bacteria (i.e., Thauera, Enterobacter and Pseudomonas) and electroactive bacteria (i.e., Geobacter) in the B-MBBR. Meanwhile, based on the content of coenzyme I (NADH) and denitrifying enzymes, biochar would also enhance electron transport activity for denitrification. Consequently, these facial prepared biocarriers are effective to enhance denitrification performance in MBBR with application significance.

Rumen Methanogenesis, Rumen Fermentation, and Microbial Community Response to Nitroethane, 2-Nitroethanol, and 2-Nitro-1-Propanol: An In Vitro Study.

Simple SummaryThe present study comparatively investigates the inhibitory difference of nitroethane (NE), 2-nitroethanol (NEOH), and 2-nitro-1-propanol (NPOH) on in vitro rumen fermentation, microbial populations, and coenzyme activities associated with methanogenesis. The results showed that both NE and NEOH were more effective in reducing ruminal methane (CH4) production than NPOH. This work provides evidence that NE, NEOH, and NPOH were able to inhibit methanogen population and dramatically decrease methyl-coenzyme M reductase gene expression and the content of coenzymes F420 and F430 with different magnitudes in order to reduce ruminal CH4 production.Nitroethane (NE), 2-nitroethanol (NEOH), and 2-nitro-1-propanol (NPOH) were comparatively examined to determine their inhibitory actions on rumen fermentation and methanogenesis in vitro. Fermentation characteristics, CH4 and total gas production, and coenzyme contents were determined at 6, 12, 24, 48, and 72 h incubation time, and the populations of ruminal microbiota were analyzed by real-time PCR at 72 h incubation time. The addition of NE, NEOH, and NPOH slowed down in vitro rumen fermentation and reduced the proportion of molar CH4 by 96.7%, 96.7%, and 41.7%, respectively (p < 0.01). The content of coenzymes F420 and F430 and the relative expression of the mcrA gene declined with the supplementation of NE, NEOH, and NPOH in comparison with the control (p < 0.01). The addition of NE, NEOH, and NPOH decreased total volatile fatty acids (VFAs) and acetate (p < 0.05), but had no effect on propionate concentration (p > 0.05). Real-time PCR results showed that the relative abundance of total methanogens, Methanobacteriales, Methanococcales, and Fibrobacter succinogenes were reduced by NE, NEOH, and NPOH (p < 0.05). In addition, the nitro-degradation rates in culture fluids were ranked as NEOH (−0.088) > NE (−0.069) > NPOH (−0.054). In brief, the results firstly provided evidence that NE, NEOH, and NPOH were able to decrease methanogen abundance and dramatically decrease mcrA gene expression and coenzyme F420 and F430 contents with different magnitudes to reduce ruminal CH4 production.

The inhibitory action mode of nitrocompounds on in vitro rumen methanogenesis: a comparison of nitroethane, 2-nitroethanol and 2-nitro-1-propanol

AbstractNitroethane (NE), 2-nitroethanol (NEOH) and 2-nitro-1-propanol (NPOH) were investigated in order to determine their inhibitory effects on in vitro ruminal fermentation and methane (CH4) production of a hay-rich substrate (alfalfa hay: maize meal = 4:1, w/w). The rumen liquor collected from cannulated Holstein dairy cows was incubated at 39 °C for 72 h. The addition of NE, NEOH and NPOH slowed down the fermentation process and notably decreased molar CH4 proportion by 96.8, 96.4 and 35.0%, respectively. The abundance of total methanogen and methanogens from the order Methanobacteriales were all decreased with NE, NEOH and NPOH supplementation. Meanwhile, the nitrocompound addition reduced mcrA gene expression, coenzyme F420 and F430 contents. The correlation analysis showed that CH4 production was correlated positively with the population abundance of total methanogens, Methanobacteriales, mcrA gene expression, coenzyme contents of F420 and F430. The nitrocompound addition decreased acetate concentration and increased propionate and butyrate concentrations in the culture fluid. In summary, both NE and NEOH addition presented nearly the same inhibitory effectiveness on in vitro CH4 production; they were more effective than NPOH. The results of the current study provide evidence that NE, NEOH and NPOH can dramatically decrease methanogen population, mcrA gene expression and the coenzyme content of F420 and F430 in ruminal methanogenesis.

The role of nicotinamide in the correction of renal function in diabetic nephropathy

Hyperglycemia of diabetes mellitus leads to the activation of the polyol way of oxidation of glucose with the activation of the enzymes of aldose reductase and sorbitol dehydrogenase and of their coenzymes NADPH and NAD, which triggers the mechanism of formation of sorbitol. The consequences of these changes lead to microangiopathy of the tissues of the kidneys, which may be one of the pathogenetic mechanisms of diabetic nephropathy. In an accessible literature, the role of coenzymes of sorbitol pathway in the development of diabetic nephropathy is not sufficiently defined. The purpose of the study was to study the content of NAD and NADPH coenzymes, their correlation, and their role in the mechanism of kidney damage in diabetes mellitus and to predict the possible correction of these changes with the NAD-nicotinamide derivative. The study was conducted on a model of streptotrozectinic diabetes mellitus (single administration of streptozotocin in a dose of 60 mg per 1 kg of body weight). Four weeks after induction of diabetes, nicotinamide (100 mg per 1 kg body weight) was injected. The level of glucose was determined by the Accu-chek (Roshe Diagnostics, Switzerland) glucose meter. The content of NAD and NADH was determined in the non-protein extracts. The statistical analysis was carried out using the Microsoft Excel statistical analysis program. The difference between the indicators was considered statistically significant (p&lt;0.05). The NAD level was reduced by 31%, the NAD/NADN ratio was 32%. The dependence of the ratio of NADP/NADPN in conditions of hyperglycemia of diabetes mellitus with clinical manifestations of diabetic nephropathy is determined. A decrease in the ratio of NADP/NADPN to 38% in the rat kidney in the cortical layer was detected. The introduction of nicotinamide normalized the reduced content of NAD diabetic rats. These results provide perspectives for further research in which nicotinamide can be used as a renal protector.

Comprehensive Profile of Acute Mitochondrial Dysfunction in a Preclinical Model of Severe Penetrating TBI.

Mitochondria constitute a central role in brain energy metabolism, and play a pivotal role in the development of secondary pathophysiology and subsequent neuronal cell death following traumatic brain injury (TBI). Under normal circumstances, the brain consumes glucose as the preferred energy source for adenosine triphosphate (ATP) production over ketones. To understand the comprehensive picture of substrate-specific mitochondrial bioenergetics responses following TBI, adult male rats were subjected to either 10% unilateral penetrating ballistic-like brain injury (PBBI) or sham craniectomy (n = 5 animals per group). At 24 h post-injury, mitochondria were isolated from pooled brain regions (frontal cortex and striatum) of the ipsilateral hemisphere. Mitochondrial bioenergetics parameters were measured ex vivo in the presence of four sets of metabolic substrates: pyruvate+malate (PM), glutamate+malate (GM), succinate (Succ), and β-hydroxybutyrate+malate (BHBM). Additionally, mitochondrial matrix dehydrogenase activities [i.e., pyruvate dehydrogenase complex (PDHC), alpha-ketoglutarate dehydrogenase complex (α-KGDHC), and glutamate dehydrogenase (GDH)] and mitochondrial membrane-bound dehydrogenase activities [i.e., electron transport chain (ETC) Complex I, II, and IV] were compared between PBBI and sham groups. Furthermore, mitochondrial coenzyme contents, including NAD(t) and FAD(t), were quantitatively measured in both groups. Collectively, PBBI led to an overall significant decline in the ATP synthesis rates (43–50%; *p < 0.05 vs. sham) when measured using each of the four sets of substrates. The PDHC and GDH activities were significantly reduced in the PBBI group (42–53%; *p < 0.05 vs. sham), whereas no significant differences were noted in α-KGDHC activity between groups. Both Complex I and Complex IV activities were significantly reduced following PBBI (47–81%; *p < 0.05 vs. sham), whereas, Complex II activity was comparable between groups. The NAD(t) and FAD(t) contents were significantly decreased in the PBBI group (27–35%; *p < 0.05 vs. sham). The decreased ATP synthesis rates may be due to the significant reductions in brain mitochondrial dehydrogenase activities and coenzyme contents observed acutely following PBBI. These results provide a basis for the use of “alternative biofuels” for achieving higher ATP production following severe penetrating brain trauma.

Effect of electroacupuncture at "Shenmen"(HT7) and "Sanyinjiao"(SP6) on energy metabolism in paraventricular nucleus of hypothalamus of insomnia rats

To observe the effect of electroacupuncture(EA) at "Shenmen"(HT7) and "Sanyinjiao"(SP6)on energy metabolism in paraventricular nucleus (PVN) of hypothalamus in insomnia rats, so as to explore its mechanism underlying improvement of insomnia. A total of 66 SD rats (half male and half female) were randomized into 3 groups：normal control, model and EA groups(n＝22 per group). The insomnia model was established by binding the rat for at least 4 h (step increase of 30 min per day), once daily for 15 days. EA (5 Hz /25 Hz, 0.5－1.0 mA) was applied to unilateral HT7 and SP6 for 15 min, once daily for 5 days. The rats' spontaneous activities during day and night were recorded by using the ClockLab Data Collection and Analysis System, and the duration of exhausted swimming was detected by using load-bearing endurance swimming test. The expression of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) of PVN tissue was assayed by Western blot, and the contents of acetyl coenzyme A (Ac-CoA) and Na＋-K＋adenosine triphosphatase (Na＋-K＋-ATPase) in the PVN tissue, and corticosterone (CORT) in plasma were assayed by ELISA. Changes of the ultrastructure of PVN cells were observed by transmission electron microscope. After modeling, the rats' daytime and nocturnal locomotor activities were significantly increased and decreased, respectively (P<0.05), and the duration of exhausted swimming was considerably shortened in the model group compared with that of the normal control group (P<0.05). The expression level of AMPK protein in the PVN was obviously up-regulated (P<0.05), and the contents of Ac-CoA and Na＋-K＋-ATPase in PVN and CORT in plasma were markedly decreased in the model control group relevant to the normal group (P<0.05). After EA intervention, the increased daytime locomotion and the decreased nocturnal activities, the shortened duration of exhausted swimming, the up-regulated expression of AMPK, and the decreased Ac-CoA, Na＋-K＋-ATPase and CORT contents were all reversed in the EA treated rats relevant to those of the insomnia rats (all P<0.05). Moreover, ultrastructural observation showed mitochondrial swelling and disappearance of partial ribosomes in the plasma of PVN cells in the model group, while in the EA group, only mild swelling of some mitochondria was found, being with basically normal nuclear membrane, mitochondria, rough endoplasmic reticulum, Golgi complex and ribosomes. EA at HT7 and SP6 has a positive effect in improving insomnia and insomnia-induced fatigue in insomnia rats, which may be associated with its effects in restraining the expression of AMPK protein, and up-regulating the contents of Ac-CoA and Na＋-K＋-ATPase in PVN and CORT in plasma.

Abstract 536: Increased O-linked Glycosylation in Diabetic Myocardium of Mice and Human

Objectives: The modification of serine and threonine residues of nuclear and cytoplasmic proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) has emerged as a highly dynamic post-translational modification that plays a critical role in regulating numerous biological processes. Our focus was to establish the difference of O-linked glycosylation in diabetic (DM) and Non-diabetic (ND) heart of mouse and human in the presence or absence of cardioplegic ischemia and cardiopulmonary bypass (CP/CPB). Methods and results: Right atrial tissue was harvested pre- and post-CP/CPB from the ND patients and patients with type-2 DM undergoing cardiac surgery (n = 4/group).The post-CP/CPB tissue samples were from a atrial tissue segment exposed to cold, hyperkalemic blood cardioplegia (ischemia) and a brief period (10 minutes) of reperfusion. Ventricular tissue samples were also obtained from mice with and without type-2 DM (n =3/group). Thin Layer Chromatography (TLC) was used to measure the contents of acetyl coenzyme A (acetyl-CoA) in the tissue lysates of mouse and human myocardium. Western blotting and immunohistochemistry analysis were employed to determine the expression/localization of O-Linked β-N-acetylglucosamine (O-GlcNAc) in the tissue lysates and paraffin-embedded tissue sections. Diabetes tends to increase the amounts of acetyl-CoA and O-GlcNAc (20% increase vs. ND, P&lt;0.05) in the myocardium compared with ND. Post-CP/CPB further induced more acetyl-CoA and this effect was more pronounced in the diabetic atrial myocardium than that of ND. Immunofluorescence experiments with O-GlcNAc-specific antibody and paraffin-embedded tissue sections from human atria and mouse heart show a significant number of cells stain positively with the antibody in the diabetes sections (3 folds increases in optical density vs. ND, P &lt;0.01). The protein oxidation and ratio of NADH/NAD in the DM myocardium were significantly increased compared with ND (P&lt;0.05). Conclusion: Diabetes and cardioplegic ischemia are associated with an increase in O-linked glycosylation in the mouse and human myocardium, suggesting that diabetes and cardioplegic ischemia affect myocardial metabolism via hexosamine biosynthetic pathway.

Fluorescence lifetime imaging of red yeast Cystofilobasidium capitatum during growth

Abstract Red yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.

Nitric oxide induces monosaccharide accumulation through enzyme S-nitrosylation.

Nitric oxide (NO) is extensively involved in various growth processes and stress responses in plants; however, the regulatory mechanism of NO-modulated cellular sugar metabolism is still largely unknown. Here, we report that NO significantly inhibited monosaccharide catabolism by modulating sugar metabolic enzymes through S-nitrosylation (mainly by oxidizing dihydrolipoamide, a cofactor of pyruvate dehydrogenase). These S-nitrosylation modifications led to a decrease in cellular glycolysis enzymes and ATP synthase activities as well as declines in the content of acetyl coenzyme A, ATP, ADP-glucose and UDP-glucose, which eventually caused polysaccharide-biosynthesis inhibition and monosaccharide accumulation. Plant developmental defects that were caused by high levels of NO included delayed flowering time, retarded root growth and reduced starch granule formation. These phenotypic defects could be mediated by sucrose supplementation, suggesting an essential role of NO-sugar cross-talks in plant growth and development. Our findings suggest that molecular manipulations could be used to improve fruit and vegetable sweetness.

Digestion performance and microbial community in full-scale methane fermentation of stillage from sweet potato-shochu production

Sweet potato shochu is a traditional Japanese spirit produced mainly in the South Kyushu area in Japan. The amount of stillage reaches approximately 8 × 105 tons per year. Wastewater mainly containing stillage from the production of sweet potato-shochu was treated thermophilically in a full-scale treatment plant using fixed-bed reactors (8 reactors × 283 m3). Following the addition of Ni2+ and Co2+, the reactors have been stably operated for six years at a high chemical oxygen demand (COD) loading rate of 14 kg/(m3·day). Analysis of coenzyme content and microbial communities indicated that similar microbial communities were present in the liquid phase and on the fiber carriers installed in reactors. Bacteria in the phyla Firmicutes as well as Bacteroidetes were dominant bacteria, and Methanosarcina thermophila as well as Methanothermobacter crinale were dominant methanogens in the reactors. This study reveals that stillage from sweet potato-shochu production can be treated effectively in a full-scale fixed-bed reactor under thermophilic conditions with the help of Ni2+ and Co2+. The high diversity of bacterial community and the coexistence of both aceticlastic and hydrogenotrophic methanogens contributed to the excellent fermentation performance.

Cd2+ resistance mechanisms in Methanosarcina acetivorans involve the increase in the coenzyme M content and induction of biofilm synthesis

To assess what defence mechanisms are triggered by Cd(2+) stress in Methanosarcina acetivorans, cells were cultured at different cadmium concentrations. In the presence of 100 μM CdCl2, the intracellular contents of cysteine, sulfide and coenzyme M increased, respectively, 8, 27 and 7 times versus control. Cells incubated for 24 h in medium with less cysteine and sulfide removed up to 80% of Cd(2+) added, whereas their cysteine and coenzyme M contents increased 160 and 84 times respectively. Cadmium accumulation (5.2 μmol/10-15 mg protein) resulted in an increase in methane synthesis of 4.5 times in cells grown on acetate. Total phosphate also increased under high (0.5 mM) Cd(2+) stress. On the other hand, cells preadapted to 54 μM CdCl2 and further exposed to > 0.63 mM CdCl2 developed the formation of a biofilm with an extracellular matrix constituted by carbohydrates, DNA and proteins. Biofilm cells were able to synthesize methane. The data suggested that increased intracellular contents of thiol molecules and total phosphate, and biofilm formation, are all involved in the cadmium resistance mechanisms in this marine archaeon.

Disruption of the histidine triad nucleotide-binding hint2 gene in mice affects glycemic control and mitochondrial function

The histidine triad nucleotide-binding (HINT2) protein is a mitochondrial adenosine phosphoramidase expressed in the liver and pancreas. Its physiological function is unknown. To elucidate the role of HINT2 in liver physiology, the mouse Hint2 gene was deleted. Hint2(-/-) and Hint2(+/+) mice were generated in a mixed C57Bl6/J × 129Sv background. At 20 weeks, the phenotypic changes in Hint2(-/-) relative to Hint2(+/+) mice were an accumulation of hepatic triglycerides, decreased tolerance to glucose, a defective counter-regulatory response to insulin-provoked hypoglycemia, and an increase in plasma interprandial insulin but a decrease in glucose-stimulated insulin secretion and defective thermoregulation upon fasting. Leptin messenger RNA (mRNA) in adipose tissue and plasma leptin were elevated. In mitochondria from Hint2(-/-) hepatocytes, state 3 respiration was decreased, a finding confirmed in HepG2 cells where HINT2 mRNA was silenced. The linked complex II-III electron transfer was decreased in Hint2(-/-) mitochondria, which was accompanied by a lower content of coenzyme Q. Hypoxia-inducible factor-2α expression and the generation of reactive oxygen species were increased. Electron microscopy of mitochondria in Hint2(-/-) mice aged 12 months revealed clustered, fused organelles. The hepatic activities of 3-hydroxyacyl-coenzyme A dehydrogenase short chain and glutamate dehydrogenase (GDH) were decreased by 68% and 60%, respectively, without a change in protein expression. GDH activity was similarly decreased in HINT2-silenced HepG2 cells. When measured in the presence of purified sirtuin 3, latent GDH activity was recovered (126% in Hint2(-/-) versus 83% in Hint2(+/+) ). This suggests a greater extent of acetylation in Hint2(-/-) than in Hint2(+/+) . Hint2/HINT2 positively regulates mitochondrial lipid metabolism and respiration and glucose homeostasis. The absence of Hint2 provokes mitochondrial deformities and a change in the pattern of acetylation of selected proteins.

Myocardial ATGL Overexpression Decreases the Reliance on Fatty Acid Oxidation and Protects against Pressure Overload-Induced Cardiac Dysfunction

Alterations in myocardial triacylglycerol content have been associated with poor left ventricular function, suggesting that enzymes involved in myocardial triacylglycerol metabolism play an important role in regulating contractile function. Myocardial triacylglycerol catabolism is mediated by adipose triglyceride lipase (ATGL), which is rate limiting for triacylglycerol hydrolysis. To address the influence of triacylglycerol hydrolysis on myocardial energy metabolism and function, we utilized mice with cardiomyocyte-specific ATGL overexpression (MHC-ATGL). Biochemical examination of MHC-ATGL hearts revealed chronically reduced myocardial triacylglycerol content but unchanged levels of long-chain acyl coenzyme A esters, ceramides, and diacylglycerols. Surprisingly, fatty acid oxidation rates were decreased in ex vivo perfused working hearts from MHC-ATGL mice, which was compensated by increased rates of glucose oxidation. Interestingly, reduced myocardial triacylglycerol content was associated with moderately enhanced in vivo systolic function in MHC-ATGL mice and increased isoproterenol-induced cell shortening of isolated primary cardiomyocytes. Most importantly, MHC-ATGL mice were protected from pressure overload-induced systolic dysfunction and detrimental structural remodeling following transverse aortic constriction. Overall, this study shows that ATGL overexpression is sufficient to alter myocardial energy metabolism and improve cardiac function.

Preservation of micronutrients during rapeseed oil refining: A tool to optimize the health value of edible vegetable oils? Rationale and design of the Optim'Oils randomized clinical trial

Numerous micronutrients naturally abundant in oilseeds prevent the risk of cardiovascular diseases by reducing cholesterolemia and oxidative stress. These micronutrients include phytosterols and various antioxidants such as polyphenols, tocopherols and coenzyme Q10/Q9 but most of them are lost during the oilseed oil refining. The main objective of the Optim'Oil project was to modify the processes of oil refining in order to reduce the lost of micronutrients. Two clinical trials (cross-over, monocentric, randomized, double-blind and controlled) were designed to investigate the effect of an optimized rapeseed oil 1) on cardiovascular biomarkers (long-term study) and 2) on oxidative stress parameters (post-prandial study). For the long-term study, 59 volunteers ingested daily 20 g of oil and 22 g of margarine (optimized or standard) for 2 periods of 3 weeks separated by a 3-week wash-out period. Blood samples were collected at the beginning and at the end of each period. For the post-prandial study, a sub-group of 16 volunteers came fasted at the laboratory and took 300 mL of a test meal containing 60% of the optimized or standard oils. Blood samples were collected before and during 6 h after the test meal intake. In comparison with the standard oil and margarine, the optimized oil and margarine exhibit as expected an increased content of phytosterol (+ 22%), polyphenols (×11), tocopherols (+ 131%) and coenzyme Q10/Q9 (+ 165%). Overall, conditions of this study were relevant to investigate the effect of the optimized rapeseed oil and margarine on the cardiovascular risk and the oxidative stress.

Kinetic model of asymmetric reduction of 3-oxo-3-phenylpropionic acid ethyl ester using Saccharomyces cerevisiae CGMCC No.2266

The kinetic model of asymmetric reduction of 3-oxo-3-phenylpropionic acid ethyl ester using Saccharomyces cerevisiae CGMCC No.2266 with 10% glucose as co-substrate to realize the regeneration of NADPH was established. The effect factors on reduction, the type and the content of co-substrate and coenzyme, and the changes of the substrate and product content vs. time during the reaction process were investigated. The results indicate that 10% glucose can increase the reaction conversion from 23.0% to 98.4% and NADPH is reducer. The reduction process conforms with sequence mechanisms. The model parameters are as follows: vm=5.0×10−4 mol·L−1·h−1, k1=1.5×10−6 mol·L−1·h−1, k2=3.0×10−3 mol·L−1·h−1. The kinetic model is in good agreement with the experimental data.

INTRAVENOUS BOLUS OF SOLUBILIZED COENZYME Q10 INCREASED RAPIDLY ITS MYOCARDIAL AND BRAIN LEVEL: PP.13.483

Objective: Coenzyme Q10 (CoQ10) is recommended for long-term treatment as hypotensive and cardioprotective agent. Bioavailability of coenzyme Q10 (CoQ10) after per os administration is very poor. To facilitate the uptake of CoQ10 by peripheral tissues is necessary to raise its plasma level many times. The aim of the study was to examine the tissues distribution of CoQ10 following intravenous bolus administration of its solubilised form. Design and Methods: The experiments were carried out on male Wistar rats with venous and arterial catheters. Solubilized CoQ10 (Kudesan solution, “Akvion”, Russia) was administrated intravenously (30 mg/kg). The content of coenzymes Q was determined by HPLC with electrochemical detection in the serum, homogenates of myocardium, brain and liver in three groups of rats: 1) 30 min group (n = 5) – 30 min after i.v. bolus; 2) 48 h group (n = 5) – 48 h after i.v. bolus and 3) control rats (n = 7). Statistical analysis was performed with two-tailed t-test. Results: Administration of solubilized CoQ10 led to enhance in its tissue concentrations. Plasma level of CoQ10 dramatically increased by 2000-fold in 30 min and by 10-fold in 48 h after bolus. Myocardial level of CoQ10 was raised by 20% (p < 0.05) in 30 min and by 98% (p < 0.01) in 48 h versus control rats. CoQ9 myocardial level also increased by 10% (p < 0.01) in 30 min and by 14% in 48 h. In the brain increased CoQ10 content was found only in 30 min after administration (by 48% versus control rats, p < 0.0001). CoQ9 brain level was not changed compared with control group. CoQ10 level in the liver in 30 min and 48 h after administration was 4,5-fold and 37-fold higher, respectively, than those in the control rats. CoQ9 liver level was decreased by 57% in 30 min and was not different from control rats in 48 h. Conclusion: Iintravenous injection of solubilised CoQ10 ensures rapid growth of tissue CoQ10 content and gives occasion to investigation of CoQ10 efficiency at acute cardiovascular events.