System biology of alcoholism: Understanding of the consequences of the metabolism in brain cells

The addictive drug ethanol represents a serious universal health and social problem. Alcohol abuse and dependency have been increasing in many countries worldwide. Chronic use of alcohol leads to metabolic abnormalities and damage to the brain, which can result in severe cognitive dysfunction and addiction. Areas of the brain such as prefrontal cortex (PFC), the white matter, the hippocampus (HP), the cerebellum and the striatum are particularly vulnerable to the effects of alcohol. The striatum (ST), a region of brain closely associated with addiction mechanisms, is commonly divided into two regions, dorsal (DS) and ventral striatum (VS); the DS comprising caudate nucleus (CN) and putamen (P) while the VS is constituted mainly by nucleus accumbens (NAc). Neuroimaging analysis indicates that microstructural degradation is occurring in alcohol-affected brains and the abnormalities may be correlated with altered locomotor activity, aggressive behavior, changes in reward/aversion-related learning, deficient motor coordination, disturbances in procedural learning and memory. The NAc is strongly implicated in drug addiction but current evidence suggests that DS has also a significant role in this process. One of the aims of the project is to carry out comparative study of the sub-regions of human alcoholic striatum using two omics technologies (proteomics and metabolomics). There were no differences in the levels of dopamine (DA), 3,4-dihydrophenylacetic acid (DOPAC), serotonin (5HT), homovanillic acid (HVA), 5-hydroxyindoleacetic acid (HIAA), histamine, L-glutamate (Glu), -aminobutyric acid (GABA), tyrosine (Tyr) and tryptophan (Tryp) between the DS (CN and P) and VS (NAc) in control brains. Choline (Ch) and acetylcholine (Ach) were higher and, norepinephrine (NE) is lower, in the VS Overall. Alcohol-affected ST had lower levels of neurotransmitters except for Glu (30% higher in the alcoholic ventral striatum). Ratios of DOPAC/DA and HIAA/5HT were higher in alcohol-affected ST indicating an increase in the DA and 5HT turnover. Glutathione was significantly reduced in all three regions of alcohol-affected ST. The pattern of changes of 13 neurometabolites in alcoholic sub-regions relative to their respective control was similar. In order to investigate correlations between the alcohol-related changes in metabolites and and altered protein expressions, the author has selected a single sub-region of ST for a global proteomics study. By analysing CN tissues, 25 unique proteins were found to be differently expressed in alcohol-affected tissue 9 relative to control. From the identified proteins two were dopamine-related proteins and one a GABA-synthesizing enzyme glutamate decarboxylase (GAD) 65. Two proteins that are related to apoptosis and/or neuronal loss (BiD and amyloid-β A4 precursor protein-binding family B member 3) were increased. These results suggest that neurotransmitter metabolism and systems possibly related to neuroprotective mechanisms in both the DS (CN and P) and the VS (NAc) are significantly influenced by long-term heavy alcohol intake associated with alcoholism. Amongst the mechanisms mediating the effects of alcohol oxidative stress may have produce a particularly significant impact and could make a strong contribution to the microstructural damage. It has been known that alcohol can have a particularly damaging effect on the PFC and the HP regions of the fetus/developing/adolescent brain. The change imparted at those stages of development could be irreversible resulting in lasting deficits in a range of personality traits and impacting decision making, memory and learning. The HP is a significant place of neurogenesis and a source of neural stem cell (NSC). These can differentiate into neurons, astrocytes and oligodendrocytes and constitute building blocks of the developing central nervous system. They can also contribute to brain repair at later stages of development and, possibly, even in adulthood. On current evidence, alcohol reduces neurogenesis but little or no tangible information is available on the actual biochemistry and/or the fate of NSC. In the present experiments, the NSC obtained from rat embryos were exposed to various concentrations of ethanol (25 to 100 mM) for up to 96 hours. The cell numbers were found to be reduced in the presence of ethanol but only at the higher concentrations (50 and 100 mM). There were no apparent dramatic changes in the morphology of the cells but the numbers of neuron-like (MAP2-positive) cells were reduced by ethanol in a dose-dependent manner. In proteome analysis of alcoholic cells, a total of 28 proteins were altered in 50 mM ethanol relative to ethanol-free control. Of these proteins some were constituents of cytoskeleton, others were involved in transcription/translation, energy metabolism, signal transduction and oxidative stress. Two of the proteins identified as altered were nucleophosmin (NPM) and dead-end protein homolog 1 (DND1). These were further studied by immunological techniques in cultured neurons and astrocytes. NPM decreased and DND1 increased in both alcohol affected neurons and astrocytes cells

Could ethanol-induced alterations in the expression of glutamate transporters in testes contribute to the effect of paternal drinking on the risk of abnormalities in the offspring?

It has been known that a preconception paternal alcoholism impacts adversely on the offspring but the mechanism of the effect is uncertain. Several findings suggest that there are signalling systems in testis that are analogous to those known to be altered by alcoholism in brain. We propose that chronic alcohol affects these systems in a manner similar to that in brain. Specifically, we hypothesise that excessive alcohol may disturb glutamatergic-like signalling in testis by increasing expression of the glutamate transporter GLAST (EAAT1). We discuss ways how to test the hypothesis as well as potential significance of some of the tests as tools in the diagnostics of chronic alcoholism

Development and maturation of Philonema species (Nematoda: Philometridae) in salomid hosts with different life histories

This study was undertaken to determine the identity of Philonema oncorhynchi from anadromous sockeye and Philonema agubernaculum from non-anadromous trout, and also to test the hypothesis that the worm in salmon is dependent on hormonal stimulus from the host for synchronization of reproduction. On the basis of differences found in life cycles, cross infection and starch gel electrophoresis, P. oncorhynchi and P. agubernaculum are considered to be different species. The hypothesis was tested experimentally, using salmon pituitary extracts and synthetic stilbestrol. Pituitary extracts accelerated the production of larvae in the uterus, of the worm and stilbestrol inhibited the gonadal development of the fish, but had no apparent effect on the worms. The hypothesis was further supported by the results of transplantation of adult but non-larvigerous worms from maturing sockeye into immature trout. Larval development failed to take place in recipient hosts. As well, larvigerous worms were collected only from sexually mature fish, whether 3, 4 or 5 years old. Rapid development of the worm paralleled the rapid gonadal development of the fish during the last 6 months of the fish's life. Sexually immature sockeye had only immature worms.Science, Faculty ofZoology, Department ofGraduat

Effects of Nitrogen Application on Physiological Characteristics of Nitrate-Tolerant Mutants of Soybean

Nitrate-tolerant mutants of soybean (Glycine max 〔L.〕 Merr.), nts1116 and nts1007 and their parent cultivar Bragg were grown in field. Plants were harvested at flowering, pod elongating and pod filling stages; and analyzed for nitrogenous components and sugar. The highest total N and ammonium-N concentration were found in nodules followed by leaf, stem and root. There were small differences in the concentrations between N- and non-N-applied plants. Nts1007 showed higher concentrations in almost all plant parts than Bragg and nts1116. Allantoin-N concentrations was highest in nts1007, especially in nodule. Nitrogen application brought about the remarkable decrease of allantoin-N in stem and root. Slight differences in sugar concentrations were observed between N-applied and non -N-applied plants and among three lines. The concentrations in root and nodule of nts1007 were higher than Bragg and nts1116, indicating that a heavily nodulating mutant, nts1007 was not recognized to be restricted in energy supply for nitrogen fixation as compared to Bragg or nts1116. There were small differences in the characteristics in pods between N- and non-Napplied plants of all lines except for allantoin-N and sugar of nts1007. Allantoin-N showed higher concentration in nts1007 than in other two lines at pod elongating stage. Sugar contents showed a decreasing tendency that was distinct in nts1007 applied with nitrogen. The high allantoin-N and low sugar concentrations in pod shells of nts1007 were considered to be due to higher N fixation by superabundant nodulation