Ganglioside or sialic acid attenuates ethanol-induced decrements in locomotion, nose-poke exploration, and anxiety, but not body temperature.

1. This laboratory has previously reported that pretreatment with ganglioside, or even with its constituent, sialic acid (SA), can attenuate certain intoxicating effects of ethanol. It was important to see if these findings could be replicated, particularly by using other measures of ethanol effects. Herein we report that pretreatment with either gangliosides or SA attenuated ethanol-induced decrements in locomotion, nose-poke exploration, and anxiety, but not body temperature. 2. An ethanol dose of 4 gm/kg caused a temperature drop of about 3 degrees C, which was unaffected by any pretreatment. The onset to sleep, however, was delayed an average of 18 or 36 secs in mice pretreated with ganglioside or SA, respectively. Ethanol-only (4 gm/kg) depressed mean cumulative locomotor activity to 31% of normal, whereas the depression was 83% of normal with beef brain ganglioside pretreatment. At 2 gm/kg ethanol alone decreased nose poking in a hole-board test to 29% of normal, but the depression was only 55-63% of normal with SA or ganglioside pretreatment. In a staircase climbing anxiety test, this dose of ethanol had no effect by itself, but both ganglioside and SA pre-treatment increased climbing by 22%. Ethanol did depress rearing to only 11% of normal, whereas rearing was 51 and 99% of normal with SA and ganglioside pretreatment, respectively. In a dark-preference test, ethanol-only caused mice to spend 64% of the time in the light, compared to 31% for controls. Time in the light was only 39 and 46% with ganglioside and SA pretreatment, respectively. 3. Blood levels of ethanol were not significantly affected by pretreatment. 4. When given alone, gangliosides significantly stimulated locomotion and staircase climbing. SA significantly decreased rearing in the staircase test. Both gangliosides and SA tended to increase nose poking, number of crossings in the dark-preference test, and time in a lighted compartment. Thus, it is possible that some of the attenuation of intoxication is attributable to non-specific stimulant properties of gangliosides and SA.     

Percentile distributions of bone measurements in Iowa children: the Iowa Bone Development Study.

Four hundred twenty-eight white children (200 boys and 228 girls) ages 4.5-6.5 yr had spine, hip, and whole-body bone mineral density (BMD) and bone mineral content (BMC) measured by dual-energy X-ray absorptiometry(DXA) as part of the Iowa Bone Development Study. Anthropometric measurements, including height, weight, and body mass index (BMI) were determined for each child at the time the bone measurements were made. The age- and gender-specific height percentile based on the 2000 CDC Growth Charts (www.cdc.gov/growthcharts/) was determined for each child. These percentiles were used to classify children into four groups as defined by the 25th, 50th,and 75th percentile cutpoints. Percentile distributions were determined within each height quartile group to delineate percentiles (5th, 25th, 50th, 75th, 95th) for BMD and BMC. Gender differences in BMD and BMC were investigated before and after stratification into height groups. Boys had higher age-height-weight-adjusted means for most BMD and BMC measures except spine BMD. Bone measurements increased with height quartile, indicating that taller children have greater BMD and BMC compared to shorter children of the same age and gender. Within any given quartile,mean BMD and BMC measurements were similar for boys and girls, with the exception of hip BMD, for which values were consistently higher for boys (p < 0.05). In addition, whole-body BMC values were higher for boys in quartiles 1 and 3 (p < 0.05). These bone measures provide norms for young white children and serve as a reference for comparison with other racial and ethnic groups, as well as with childhood populations that are at risk for osteopenia because of chronic disease. Gender, age, and height are useful clinical predictors of BMD and BMC in young children.     

Chlorambucil-monoglutathionyl conjugate is sequestered by human alpha class glutathione S-transferases.

The spontaneous reaction of 110 microM chlorambucil (4-[p-[bis(2-chloroethyl)amino]phenyl]-butanoic acid; CHB) with 5 mM GSH at 37 degrees C in physiological phosphate-buffered saline for 35 min gave primarily the monoglutathionyl derivative, 4-[p-[N-2-chloroethyl,N-2-S-glutathionylethyl]amino]phenyl]-butano ic acid; CHBSG) and the diglutathionyl derivative, 4-[p-[bis(2-S-glutathionylethyl]amino]phenyl]-butanoic acid (CHBSG2) with small amounts of the hydroxy-derivatives: 4-[p-[N-2-chloroethyl,N-2-hydroxy-ethyl]amino] phenyl-butanoic acid (CHBOH) and 4-[p-[N-2-S-glutathionylethyl-2-hydroxyethyl]amino]phenyl]-butanoi c acid (CHBSGOH). The inclusion of approximately physiological amounts of human glutathione S-transferases (GSTs) A1-1, A2-2, P1-1, M1a-1a M3-3 or P1-1 (for nomenclature see Mannervik et al., 1992, Biochem. J., 282, 305) had little or no catalytic effect on these reactions as determined by loss of CHB. However, GTSs A1-1 and A2-2 were associated with a significant increase of CHBSG at the expense of CHBSG2 + CHBSGOH suggesting that these GTs sequestered CHBSG at the active site. This interpretation was supported by inhibition studies which showed that CHBSG was a pure competitive inhibitor of the activity of GSTs A1-1 and A2-2 towards 1-chloro-2,4-dinitrobenzene with Ki's of 1.3 and 1.2 microM respectively. GSH transferases P1-1 and M1a-1a were inhibited by CHBSG above 10 microM. Incubation of 2 microM CHB, a concentration which may be of more significance for chemotherapy, in the presence or absence of GST A1-2 (20-50 microM) showed catalysis of GSH monoconjugation equivalent to 18% of the spontaneous rate. However, the dominant effect again was the sequestration of CHBSG which reached 74.3 +/- 1.5 (SEM)% of the total reactants at 60 min compared to 28.9 +/- 0.3(SEM)% in controls. CHBSG, although possessing a potential electrophilic centre, showed no detectable alkylation of plasmid DNA but indirect evidence was obtained that it alkylated other cellular macromolecules. It is concluded that the contribution of GSTs to catalysis of CHB detoxication will depend on factors not previously considered, namely the relative molarities of CHB, CHBSG and GSTs, and the cellular capacity to excrete CHBSG to relieve product inhibition.     

Temporary adhesions between axons and myelin-forming processes

Following irradiation, the dorsal funiculus of the lumbosacral spinal cord in the rat undergoes the following sequence of events: (a) a marked reduction of the normal glial population, (b) an absence of oligodendrocyte myelin formation, (c) the invasion and proliferation of Schwann cells, and (d) the myelination of axons within the cord by Schwann cells. The present study demonstrates that, during the latter process, junctional complexes develop between these intraspinal Schwann cells and the axolemma. These complexes are present at sites of probable initial contact between the two membranes. As the Schwann cell process begins to wrap the axons, these junctional complexes are located between the inner spiraling process of the Schwann cell and the axon. With the advancement of myelin formation to the stage of 8 to 9 compact spirals, these contacts are rarely observed. Spinal cords from normal 8-day-old rats were examined in order to determine if such contacts occur during myelination by oligodendrocytes. Although they are more difficult to detect in the normal animal due to the abundance of glial processes, similar junctional complexes occur between oligodendrocyte processes and axons. These observations suggest that these complexes may serve to stabilize and to guide the myelin-forming process around the perimeter of the axon. Additionally, these junctions may play an active role in the advancement of the inner spiraling process by forming temporary adhesions between the axolemma and the adjacent myelin-forming process. Coated vesicles are commonly observed fused with the axolemma of axons which are in the early stages of myelination. These coated vesicles may be involved in the insertion or the deletion of junctional membrane.