Effects of ventral striatal 6-OHDA lesions or amphetamine sensitization on ethanol consumption in the rat

Female rats with continuous access to water and 6% ethanol were given bilateral ventral striatal 6-OHDA infusions, which induced pronounced striatal depletions of dopamine. The postoperative ethanol consumption of these rats was not significantly affected in comparison to vehicle-infused controls. In a second experiment, female rats received escalating doses of d-amphetamine over a 5-week period (from 1 to 9 mg/kg/injection). Control females were given saline injections. Following a 3-month drug-free interval, the females were given access to ethanol, the concentration of which was gradually increased from 2% to 12% with weekly intervals. Amphetamine-sensitized rats consumed significantly more alcohol than the saline-treated controls. Taken together, these results suggest that striatal dopaminergic mechanisms, while not necessary for basal ethanol drinking, can facilitate alcohol drinking.     

The role of catecholamines in desmopressin induced locomotor stimulation

Central and peripheral administration of DDAVP increase locomotor activity in rats in doses that alter brain dopamine neurochemistry. In order to delineate the role of catecholamines in this behavioural effect of DDAVP, the effects of different catecholamine manipulating agents on DDAVP-induced locomotor stimulation were studied in rats. The catecholamine depleting agent reserpine (5 mg/kg), administered alone or together with the catecholamine synthesis inhibitor alpha-methyltyrosine (250 mg/kg), completely prevented the locomotor stimulatory effect of DDAVP. The dopamine D1 receptor antagonist Sch-23390 (0.01 and 0.03 mg/kg) significantly antagonized the DDAVP-induced locomotor stimulation when administered in the higher dose, that also produced a significant reduction of locomotor activity per se, whereas the dopamine D2 receptor antagonist raclopride (0.08 and 0.16 mg/kg) had no significant effect. The two dopamine blockers administered together produced a significant, dose-dependent reduction of DDAVP-induced locomotor stimulation, while controls were not significantly affected. Also the alpha-adrenoceptor antagonist phenoxybenzamine decreased the DDAVP-induced locomotor stimulation in a dose (20 mg/kg) that did not influence locomotor activity in controls, and, finally, administration of Sch-23390, raclopride and phenoxybenzamine antagonised the DDAVP-induced effect in a dose combination that failed to influence locomotor activity per se. In vivo microdialysis experiments in awake, freely moving rats indicated that DDAVP increases dopamine overflow in the nucleus accumbens, a brain area of importance for initiation of locomotor activity, by approximately 25%, as compared to baseline levels. Taken together, these results indicate that the central stimulatory action of DDAVP involves granula-mediated dopamine release and subsequent activation of dopamine D1 and D2 receptors, and that alpha-adrenoceptors possibly also are involved.     

Cysteine protease inhibitors cure an experimental Trypanosoma cruzi infection

Trypanosoma cruzi is the causative agent of Chagas' disease. The major protease, cruzain, is a target for the development of new chemotherapy. We report the first successful treatment of an animal model of Chagas' disease with inhibitors designed to inactivate cruzain. Treatment with fluoromethyl ketone-derivatized pseudopeptides rescued mice from lethal infection. The optimal pseudopeptide scaffold was phenylalanine-homophenylalanine. To achieve cure of infection, this pseudopeptide scaffold was incorporated in a less toxic vinyl sulfone derivative. N-methyl piperazine-Phe-homoPhe-vinyl sulfone phenyl also rescued mice from a lethal infection. Six of the treated mice survived over nine months, three without further treatment. Three mice that had entered the chronic stage of infection were retreated with a 20-d regimen. At the conclusion of the experiments, five of the six mice had repeated negative hemacultures, indicative of parasitological cure. Studies of the effect of inhibitors on the intracellular amastigote form suggest that the life cycle is interrupted because of inhibitor arrest of normal autoproteolytic cruzain processing at the level of the Golgi complex. Parasites recovered from the hearts of treated mice showed the same abnormalities as those treated in vitro. No abnormalities were noted in the Golgi complex of host cells. This study provides proof of concept that cysteine protease inhibitors can be given at therapeutic doses to animals to selectively arrest a parasitic infection.     

Crystal structure of enoyl-coenzyme A (CoA) hydratase at 2.5 angstroms resolution: a spiral fold defines the CoA-binding pocket

The crystal structure of rat liver mitochondrial enoyl-coenzyme A (CoA) hydratase complexed with the potent inhibitor acetoacetyl-CoA has been refined at 2.5 angstroms resolution. This enzyme catalyses the reversible addition of water to alpha,beta-unsaturated enoyl-CoA thioesters, with nearly diffusion-controlled reaction rates for the best substrates. Enoyl-CoA hydratase is a hexamer of six identical subunits of 161 kDa molecular mass for the complex. The hexamer is a dimer of trimers. The monomer is folded into a right-handed spiral of four turns, followed by two small domains which are involved in trimerization. Each turn of the spiral consists of two beta-strands and an alpha-helix. The mechanism for the hydratase/dehydratase reaction follows a syn-stereochemistry, a preference that is opposite to the nonenzymatic reaction. The active-site architecture agrees with this stereochemistry. It confirms the importance of Glu164 as the catalytic acid for providing the alpha-proton during the hydratase reaction. It also shows the importance of Glu144 as the catalytic base for the activation of a water molecule in the hydratase reaction. The comparison of an unliganded and a liganded active site within the same crystal form shows a water molecule in the unliganded subunit. This water molecule is bound between the two catalytic glutamates and could serve as the activated water during catalysis.     

Predicting optimal time of insemination in cows that show visual signs of estrus by estimating onset of estrus with pedometers

An experiment was designed to estimate the optimal interval from the beginning of estrus to artificial insemination (AI). The data were analyzed by means of a mathematical model. The analysis was based on pedometer readings and results of rectal palpation at 42 to 49 d post-AI of 171 breedings in 121 cows. The chance of conception was highest between 6 and 17 h after increased pedometer activity; the estimated optimum was at 11.8 h. In this data file, the effects of disease, inseminator, time of AI (a.m. or p.m.), and bull did not contribute to the improvement of the model. The effects of disease were not significant because of the low incidence of any specific disease. Activity measurements can be used as a tool for AI strategy to improve conception in groups of healthy cows and heifers already showing visual signs of estrus.     

Schnelle Mikromethoden zur GC- Bestimmung des Fettgehalts und der Fettsauren in lipdereichen Samen

Rapid micro methods for the GLC determination of fat content and fatty acid composition in seeds rich in lipids. Two methods for the rapid sample preparation for determination of fat (gravimetri cally resp. by GLC) and fatty acid pattern (by GLC) in seeds with high lipid contents arc presented and discussed. Tire adequate amount of the sample is no more than 50 mg for both methods. The first method requires several subsequent steps: grinding with Na₂S0₄ and petroleum benzine, fat determination by weighing. transmethylation by sodium methylate in methanol (time required: approx. 60 min.). The second method is an one-step method: grinding with Na₂SO₄, extraction with petroleum benzine and transmethylation by methanol and tetramethylguanidine at the same time (time required: approx. 15 min.) – the fat content is determined by converting data of the fatty acid GLC results into data of triglycerides. The analytical results for different samples are compared with those obtained by a classical reference method. In most cases the results are comparable.     

Theoretische und experimentelle Untersuchungen zum Kriechverhalten thermisch inhomogen belasteter hohlzylindrischer Prüfkörper

Theoretical and Experimental Investigations of the Creep Behaviour of a Tube Subjected to Thermal and Mechanical Loading The inhmogeneous temperature distribution for a tubular specimen, cooled from the axial bore, has been calculated. Application of the reference value concept leads to a good approximation for the axial creep strain of the specimen. The calculations apply to steady-state as well as to transient deformations and temperature distributions. A sharp creep rate increase following abrupt changes of the temperature field is predicted; this may lead to a lifetime reduction. – The theoretical results have been checked using specimens of the alloy NiCr 80 20 in a specially designed creep machine for tubular, cooled specimens, capable of producing temperature gradients up to 50 K/mm. The agreement between theoretical and experimental results is very satisfactory, both for stationary and for transient situations.