In vitro microbial metabolism of fumonisin B1

There is a lack of information on the effect of swine caecal microbiota on fumonisin metabolism. In this in vitro study, the biotransformation of fumonisin B₁ (FB₁) by the gut microbiota of adult, healthy pigs was examined. Suspensions of caecal contents and McDougall buffer solution were incubated anaerobically with pure FB₁ for 0, 12, 24, 48 and 72 h. After 48 h, the conversion of FB₁ to partially hydrolysed FB₁ (46%) was nearly equal to the percentage ratio of FB₁, while by 72 h it was 49%. In vitro, the conversion of fumonisin B₁ to aminopentol was less than 1%. The results show that the caecal microbiota are capable of transforming fumonisin B₁ to the above metabolites. Further studies on FB₁ metabolism in the small intestine are clearly justified.     

Enhancer Regulation of Dopaminergic Neurochemical Transmission in the Striatum

The trace amine-associated receptor 1 (TAAR1) is a Gs protein-coupled, intracellularly located metabotropic receptor. Trace and classic amines, amphetamines, act as agonists on TAAR1; they activate downstream signal transduction influencing neurotransmitter release via intracellular phosphorylation. Our aim was to check the effect of the catecholaminergic activity enhancer compound ((−)BPAP, (R)-(−)-1-(benzofuran-2-yl)-2-propylaminopentane) on neurotransmitter release via the TAAR1 signaling. Rat striatal slices were prepared and the resting and electrical stimulation-evoked [³H]dopamine release was measured. The releaser (±)methamphetamine evoked non-vesicular [³H]dopamine release in a TAAR1-dependent manner, whereas (−)BPAP potentiated [³H]dopamine release with vesicular origin via TAAR1 mediation. (−)BPAP did not induce non-vesicular [³H]dopamine release. N-Ethylmaleimide, which inhibits SNARE core complex disassembly, potentiated the stimulatory effect of (−)BPAP on vesicular [³H]dopamine release. Subsequent analyses indicated that the dopamine-release stimulatory effect of (−)BPAP was due to an increase in PKC-mediated phosphorylation. We have hypothesized that there are two binding sites present on TAAR1, one for the releaser and one for the enhancer compounds, and they activate different PKC-mediated phosphorylation leading to the evoking of non-vesicular and vesicular dopamine release. (−)BPAP also increased VMAT2 operation enforcing vesicular [³H]dopamine accumulation and release. Vesicular dopamine release promoted by TAAR1 evokes activation of D2 dopamine autoreceptor-mediated presynaptic feedback inhibition. In conclusion, TAAR1 possesses a triggering role in both non-vesicular and vesicular dopamine release, and the mechanism of action of (−)BPAP is linked to the activation of TAAR1 and the signal transduction attached.     

A Constraint-Based Approach to Assigning System Components to Tasks

In multi-component systems, individual components must be assigned to the tasks that they are to perform. In many applications, there are several possible task decompositions that could be used to achieve the task, and there are limited resources available throughout the system. We present a technique for making task assignments under these conditions. Constraint satisfaction is used to assign components to particular tasks. Heuristics suggest a task decomposition for which an assignment can be found efficiently. We have applied our technique to the problem of task assignment in systems of underwater robots and instrument platforms working together to collect data in the ocean.     

GDNF Increases Inhibitory Synaptic Drive on Principal Neurons in the Hippocampus via Activation of the Ret Pathway

Glial cell line-derived neurotrophic factor (GDNF) has been shown to counteract seizures when overexpressed or delivered into the brain in various animal models of epileptogenesis or chronic epilepsy. The mechanisms underlying this effect have not been investigated. We here demonstrate for the first time that GDNF enhances GABAergic inhibitory drive onto mouse pyramidal neurons by modulating postsynaptic GABA_A receptors, particularly in perisomatic inhibitory synapses, by GFRα1 mediated activation of the Ret receptor pathway. Other GDNF receptors, such as NCAM or Syndecan3, are not contributing to this effect. We observed similar alterations by GDNF in human hippocampal slices resected from epilepsy patients. These data indicate that GDNF may exert its seizure-suppressant action by enhancing GABAergic inhibitory transmission in the hippocampal network, thus counteracting the increased excitability of the epileptic brain. This new knowledge can contribute to the development of novel, more precise treatment strategies based on a GDNF gene therapy approach.     

Global Metabolomics Discovers Two Novel Biomarkers in Pyridoxine-Dependent Epilepsy Caused by ALDH7A1 Deficiency

Pyridoxine-dependent epilepsy (PDE) is a rare autosomal recessive developmental and epileptic encephalopathy caused by pathogenic variants in the ALDH7A1 gene (PDE-ALDH7A1), which mainly has its onset in neonates and infants. Early diagnosis and treatment are crucial to prevent severe neurological sequelae or death. Sensitive, specific, and stable biomarkers for diagnostic evaluations and follow-up examinations are essential to optimize outcomes. However, most of the known biomarkers for PDE lack these criteria. Additionally, there is little discussion regarding the interdependence of biomarkers in the PDE-ALDH7A1 metabolite profile. Therefore, the aim of this study was to understand the underlying mechanisms in PDE-ALDH7A1 and to discover new biomarkers in the plasma of patients using global metabolomics. Plasma samples from 9 patients with genetically confirmed PDE-ALDH7A1 and 22 carefully selected control individuals were analyzed by ultra high performance liquid chromatography–high-resolution mass spectrometry (UHPLC-HRMS). Two novel and reliable pyridoxine-independent diagnostic markers, 6-hydroxy-2-aminocaproic acid (HACA) and an isomer of C₉H₁₁NO₄, were identified. Furthermore, a possible reaction mechanism is proposed for HACA. This study demonstrates the capability of global metabolomics in disease screening to detect established and novel biomarkers.     

Influence of Two Widely Used Solvents,Ethanol and Dimethyl Sulfoxide,on Human Sperm Parameters

To study mechanisms involved in fertility, many experimental assays are conducted by incubating spermatozoa in the presence of molecules dissolved in solvents such as ethanol (EtOH) or dimethyl sulfoxide (DMSO). Although a vehicle control group is usually included in such studies, it does not allow to evaluate the intrinsic effect of the solvent on sperm parameters and its potential influence on the outcome of the experiment. In the present study, we incubated human spermatozoa for 4 h in a capacitation medium in the absence or the presence of different concentrations of EtOH and DMSO (0.1, 0.5, 1.0, and 2.0%) to assess the impact of these solvents on sperm motility, vitality, capacitation, and acrosome integrity. The presence of statistically significant relationships between increasing solvent concentrations and the investigated parameters was assessed using linear mixed models. A significant effect was observed with both solvents for total and progressive sperm motilities. We also evaluated the effect of time for these parameters and showed that the influence of the solvents was stable between 0 and 4 h, indicating an almost direct impact of the solvents. While EtOH did not influence sperm vitality and acrosome integrity, a significant effect of increasing DMSO concentrations was observed for these parameters. Finally, regarding capacitation, measured via phosphotyrosine content, although a dose-dependent effect was observed with both solvents, the statistical analysis did not allow to precisely evaluate the intensity of the effect. Based on the results obtained in the present study, and the corresponding linear mixed models, we calculated the concentration of both solvents which would result in a 5% decline in sperm parameters. For EtOH, these concentrations are 0.9, 0.7, and 0.3% for total motility, progressive motility, and capacitation, respectively, while for DMSO they are 1.5, 1.1, >2, 0.3 and >2% for total motility, progressive motility, vitality, capacitation, and acrosome integrity, respectively. We recommend using solvent concentrations below these values to dissolve molecules used to study sperm function in vitro, to limit side effects.     

Comparison between Immunocytochemistry,FISH and NGS for ALK and ROS1 Rearrangement Detection in Cytological Samples

The detection of ROS1 and ALK rearrangements is performed for advanced-stage non-small cell lung cancer. Several techniques can be used on cytological samples, such as immunocytochemistry (ICC), fluorescence in situ hybridization (FISH) and, more recently, next-generation sequencing (NGS), which is gradually becoming the gold standard. We performed a retrospective study to compare ALK and ROS1 rearrangement results from immunocytochemistry, FISH and NGS methods from 131 cytological samples. Compared to NGS, the sensitivity and specificity of ICC were 0.79 and 0.91, respectively, for ALK, and 1 and 0.87 for ROS1. Regarding FISH, the sensitivity and specificity were both at 1 for ALK and ROS1 probes. False-positive cases obtained by ICC were systematically corrected by FISH. When using ICC and FISH techniques, results are very close to NGS. The false-positive cases obtained by ICC are corrected by FISH, and the true-positive cases are confirmed. NGS has the potential to improve the detection of ALK and ROS1 rearrangements in cytological samples; however, the cost of this technique is still much higher than the sequential use of ICC and FISH.     

Enzymatic glycerolysis and transesterification of vegetable oil for enhanced production of feruloylated glycerols

Novel functional groups can be introduced into vegetable oils using enzymes, resulting in value-added products. The transesterification kinetics of ethyl ferulate with MAG, DAG, and TAG were examined. Transesterification was catalyzed by immobilized Candida antarctica lipase B in solventless batch and packed-bed reactors. Initial reaction rates with TAG were slightly sensitive to water activity, whereas rates with MAG and DAG were water activity independent. Transesterification was also three-to sixfold faster with MAG and DAG. These observations indicate that the reaction is rate limited by the acyl acceptor, and that oils with free hydroxyl groups are preferred acyl acceptors in comparison with TAG, which must undergo partial hydrolysis before becoming reactive.     

Effect of concentration and incubation temperature on the acid induced aggregation of soymilk

The molecular details of the aggregation of soy protein particles in soymilk during acid-induced gelation using glucono-delta-lactone (GDL) were investigated. Soymilk samples were prepared from different water-to-bean ratios and contained approximately 4% and 7% protein. The effects of protein concentration and incubation temperature (30 or 7 °C) on soymilk gelation were observed using rheology and diffusing wave spectroscopy at two different GDL concentrations. During acidification, there was a decrease in electrostatic repulsion between particles which well correlated with the pH of aggregation determined by dynamic light scattering. Gelation of soymilk occurred at about pH 5.8, and neither the rate of acidification nor the protein concentration affected the gelation pH. Gel stiffness was affected by protein concentration. A detailed study of the soluble fraction during the preceding stages of aggregation clearly demonstrated that glycinin components were the first to destabilize during acidification, followed by the β-conglycinin subunits. Decreasing the incubation temperature from 30 °C to 7 °C increased the pH of gelation and the gel modulus (G′) measured by rheology. It was concluded that short range interactions play a major role in the formation of the protein network in soymilk curd.     

Optimization of hydrolysis of sardine (Sardina pilchardus) heads with Protamex: enhancement of lipid and phospholipid extraction

BACKGROUND: Fish by‐products are not considered as valuable raw materials even if they usually contain valuable components such as lipids. Fish lipids are well known for their nutritional potential and health effects but their extraction remains problematic due to the use of organic solvents. Enzymatic hydrolysis such as the proteolysis of tissues can lead to lipid extraction. RESULTS: Hydrolysis of sardine heads by Protamex was studied (temperature, hydrolysis time and enzyme–substrate ratio) using response surface methodology in order to obtain the highest release of lipids and particularly phospholipids. No relation between the degree of hydrolysis and lipid recovery were depicted; however, optimum conditions for both the release of lipids and phospholipids were found to be similar (29 min, 31 °C with 2.6 g kg^?1 enzyme). Under these hydrolysis conditions, rich lipid and phospholipid fractions (oily and aqueous fractions) can be recovered when time, temperature and enzyme consumption are minimized. Analytical data have revealed that they contain high‐quality lipids, especially ω3 fatty acid. CONCLUSION: This study demonstrated that proteolysis can be used for high lipid recovery from little‐exploited biomass like fish heads without requiring solvent or thermal treatment. Resulting phospholipids, fatty acids and peptides could be utilized for nutritional or feed purposes. Copyright © 2009 Society of Chemical Industry