Na+/K+-ATPase Inhibition Partially Mimics the Ethanol-Induced Increase of the Golgi Cell-Dependent Component of the Tonic GABAergic Current in Rat Cerebellar Granule Cells

Cerebellar granule cells (CGNs) are one of many neurons that express phasic and tonic GABAergic conductances. Although it is well established that Golgi cells (GoCs) mediate phasic GABAergic currents in CGNs, their role in mediating tonic currents in CGNs (CGN-I_tonic) is controversial. Earlier studies suggested that GoCs mediate a component of CGN-I_tonic that is present only in preparations from immature rodents. However, more recent studies have detected a GoC-dependent component of CGN-I_tonic in preparations of mature rodents. In addition, acute exposure to ethanol was shown to potentiate the GoC component of CGN-I_tonic and to induce a parallel increase in spontaneous inhibitory postsynaptic current frequency at CGNs. Here, we tested the hypothesis that these effects of ethanol on GABAergic transmission in CGNs are mediated by inhibition of the Na⁺/K⁺-ATPase. We used whole-cell patch-clamp electrophysiology techniques in cerebellar slices of male rats (postnatal day 23–30). Under these conditions, we reliably detected a GoC-dependent component of CGN-I_tonic that could be blocked with tetrodotoxin. Further analysis revealed a positive correlation between basal sIPSC frequency and the magnitude of the GoC-dependent component of CGN-I_tonic. Inhibition of the Na⁺/K⁺-ATPase with a submaximal concentration of ouabain partially mimicked the ethanol-induced potentiation of both phasic and tonic GABAergic currents in CGNs. Modeling studies suggest that selective inhibition of the Na⁺/K⁺-ATPase in GoCs can, in part, explain these effects of ethanol. These findings establish a novel mechanism of action of ethanol on GABAergic transmission in the central nervous system.     

Preparation ofN-acetylneuraminic acid from delipidated egg yolk

Egg yolk, a large proportion of the egg, was studied for the preparation ofN-acetylneuraminic acid (Neu5Ac). The delipidated hen egg yolk (DEY; 500 kg containing 0.2% w/w, Neu5Ac) was hydrolysed with HCl (pH 1.4) at 80 °C and neutralized with NaOH (pH 6.0). The mixture was filtered and electrodialysed until the conductivity was 240 µS cm^–1. The filtrate was applied on a column of Dowex HCR-W2 (20–50 mesh), followed by a column of Dowex 1-X8 (200–400 mesh). The latter column was washed with water, and then eluted with a linear gradient of HCO₂H (0–2m). The eluates containing Neu5Ac were concentrated using a reverse osmosis membrane and, finally, rotary evaporated at 40 °C. The residue was then lyophilized to yield 500 g Neu5Ac. The purity of Neu5Ac was >98% (TBA method). HPLC, NMR spectroscopy and TLC chromatography of the product obtained from the DEY showed that Neu5Ac was the sole derivative present in egg yolk. The DEY, a byproduct from egg processing plants, was found to be an excellent source for the large-scale preparation of Neu5Ac.Abbreviations Neu5Ac N-acetylneuraminic acid - Neu5Gc N-glycolylneuraminic acid - DEY delipidated egg yolk - HPLC high performance liquid chromatography - TLC thin layer chromatography - NMR nuclear magnetic resonance - IR infrared spectroscopy Presented at the 11th International Symposium on Glycoconjugates, Toronto, Canada.     

Synthesis of New Bis(pyrazolo[1,5-a]pyrimidines) Linked to Different Spacers as Potential MurB Inhibitors

An efficient protocol was adopted to efficiently prepare three new series of bis(pyrazolo[1,5-a]pyrimidines) linked to different spacers. The new bis(pyrazolo[1,5-a]pyrimidines) were prepared in 80–90 % yields by reacting the respective bis(enaminones) and 4-(4-substituted benzyl)-1H-pyrazole-3,5-diamines in pyridine at reflux temperature for 5–7 h. The new products showed a wide spectrum of antibacterial activity against six different bacterial strains. In general, propane- and butane-linked bis(pyrazolo[1,5-a]pyrimidines), which are attached to 3-(4-methyl- or 4-methoxybenzyl) units, had the best antibacterial activity with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values up to 2.5 and 5.1 μM, respectively. Additionally, the previous products demonstrated promising MurB inhibitory activity with IC₅₀ values up to 7.2 μM.     

Cycloartane triterpene glycosides from Astragalus trigonus

Three new cycloartane glycosides, trigonoside I, II and III, and the known astragalosides I and II were isolated from the roots of Astragalus trigonus. The structures of the new glycosides were totally elucidated by high field (600 MHz) NMR analyses as cycloastragenol-6-O-β-xylopyranoside, cycloastragenol-3-O-[-l-arabinopyranosyl(1 → 2)-β-d-xylopyranosyl]-6-O-β- d-xylopyranoside and cycloastragenol-3-O-[-l-arabinopyranosyl(1 → 2)-β-d-(3-O-acetyl)-xylopyranosyl]-6-O-β-d-xylopyranoside.     

Studies on antiviral glycosides. II. Mode of action for virucidal effects on Sendai virus

Treatment of Sendai virus with $\text{p-(sec-}\text{butyl)-phenyl-6-chloro-6-deoxy-β-}\text{d}\text{-glucopyranoside}$, followed by freezing and thawing resulted in a loss of hemolytic and cell fusion activities as well as infectivity without affecting hemagglutinating and neuraminidase activities. The anti-hemolytic activity of this compound was reversed by the addition of phosphatidyl choline to the virus samples. $\text{p-}\text{Azidophenyl-6-chloro-6-deoxy-β-}\text{d}\text{-[}{}^3\text{H]glucopyranoside}$ was successfully used for photoaffinity labeling of a specific virion site, and we confirmed the affected site of the glucoside to be the lipid components in the viral envelopes.     

Studies on antiviral glycosides. II. Chemical modifications of the envelope of Sendai virus

Treatment of Sendai virus with p-azidophenyl-6-chloro-6-deoxy-beta-D-glucopyranoside (APG) caused chemical modification of the viral envelope under UV irradiation, which did not affect the hemagglutinin activity of the virus but inhibited the hemolytic activity. Also, the transfer of phospholipid from the viral envelope to chicken erythrocytes was measured using a spinlabel technique by electron spin resonance (ESR). In this experiment, the phospholipid transfer was depressed by the treatment with APG under the conditions which inhibited the hemolytic activity of the virus. These results suggest that APG bound covalently to lipid may disturb the specific interaction between the protein and the lipid of the viral envelope, resulting in the inhibition of the hemolytic activity. The effects of APG on the hemolysis and phospholipid transfer were compared with the results for the concanavalin A- and amphotericin B-treated viruses.     

Cooperativity of lectin binding to lymphocytes,and its relevance to mitogenic stimulation

The relationship between the binding patterns of soybean agglutinin, peanut agglutinin (both in their native (unaggregated) form and in their polymerized form), and of Phaseolus vulgaris leucoagglutinin, to neuraminidase-treated lymphocytes from different sources, and the mitogenic activity of these lectins, was studied. In all cases investigated, binding of a lectin to lymphocytes which resulted in stimulation was a positive cooperative process. Our findings support the assumption that clustering of receptors and conformational changes in membrane structure are prerequisites for mitogenic stimulation.     

Some observations on the fine structure of the mantis shrimp heart

Summary The electron microscopic structure of the myocardium of the mantis shrimp is descriped. Particular attention is paid to the organization of the nerve terminal and the sarcotubular system. The general appearance of this myocardium is characterized by deep invagination of the plasma membrane at the level of Z-band and large irregular shaped mitochondria. It possesses a very well developed sarcotubular system, consisting of the longitudinal system and two transverse elements making two sets of contact to each other. One forms dyad and the other forms triad at the level of M- and Z-band, respectively. The organization of the myoneural junction in this muscle is very simple and undifferentiated. In general, a special structural differentiation is invariably observed at both sides of the contact area. In spite of the fact that synaptic vesicles and a differentiated membrane are found at the naked process of this cardiac nerve, specialization such as subsynaptic fold formation has not been observed at the muscle side which is in contact with the nerve process. Observations made on the sarcotubular system and the nerve termination have been discussed with reference to their physiological significance. This investigation was supported by the Public Health Service Research Grants HE 06968-04 and NB 03348-04 of the National Institutes of Health, Bethesda, and the U. S. Department of the Army through its Far East Research Office.     

Effect of deltamethrin-incorporated nets on mobility and survivorship of Halyomorpha halys (Hemiptera: Pentatomidae) adults and nymphs in the laboratory

The brown marmorated stink bug, Halyomorpha halys Stål (Hemiptera: Pentatomidae), is an invasive pest that attacks specialty and row crops in North America and Europe. There has been a concerted effort to reduce frequent broad-spectrum insecticide applications made on vulnerable crops. One tool that has emerged recently is the use of long-lasting insecticide-treated nets (LLINs) as a killing agent. Here, we conducted bioassays to evaluate the effect of direct contact on deltamethrin-impregnated LLINs on the behaviour and survivorship of H. halys nymphs and adults in the laboratory. Following exposure at three different durations (1.25, 4.25 or 7.25 min), vertical and horizontal mobility of adults and nymphs and the flight capacity of adults were recorded and compared with individuals that were not exposed (control). Exposure to LLINs reduced the horizontal distance and velocity and increased the angular velocity of adults only but reduced vertical mobility of adults and nymphs. Adult flights were not significantly affected by LLIN exposure. Mortality of adults and nymphs at 7-day post-exposure ranged from 73% to 77% regardless of exposure time. We discuss our findings within the context of the potential for and limitations of deploying LLINs in vulnerable crops to manage H. halys populations.