Rapid Communication Chronic Ingestion of Ethanol Up-Regulates NMDAR1 Receptor Subunit Immunoreactivity in Rat Hippocampus

We examined the effects of chronic ethanol exposure on the levels of N -methyl-D-aspartate receptor subunit 1 (NMDAR1) protein, an essential component of N -methyl-D-aspar- tate glutamate receptors, in rat brain. By immunoblotting procedures using a specific antibody for the NMDAR1 subunit, we found that ethanol dramatically up-regulated (by 65%) NMDAR1 immunoreactivity in the hippocampus but not in the nucleus accumbens, cerebral cortex, or striatum. In contrast, ethanol did not alter the levels of glutamate receptor subunit (GLUR) 1 or GLUR2 protein, subunits that make up the α-amino-3-hydroxy-5-methy4-isoxazole propionic acid glutamate receptor, in the hippocampus. Because ethanol can potentially influence many different neurotransmitter systems, we examined whether chronic treatment with several psychotropic drugs with different pharmacological profiles (cocaine, haloperidol, SCH 23390, imipramine, and morphine) could mimic the effect of ethanol. None of these agents increased hippocampal NMDAR1 subunit immunoreactivity after chronic administration. Increased NMDAR1 subunit levels in the hippocampus after chronic ethanol exposure may represent an important neurochemical substrate for some of the features associated with ethanol dependence and withdrawal.     

NON-RANDOM ASSOCIATION OF MITOTIC CELLS FROM CULTURED HUMAN LYMPHOCYTE SUSPENSIONS

By using chromosome markers in two separate cell lines of a human dispermic chimera, it was shown that 4.9% of metaphases in suspensions of PHA-cultured lymphocytes were paired because of non-random factors. A similar amount of metaphase pairing occurred in cultured lymphocytes of normal donors, and evidence from the relative mitotic cycles of the paired cells indicated that some of this pairing was non-random. Such non-random pairing could be a source of bias in cell kinetic and other studies involving metaphase cells in lymphocyte cultures.     

POTASSIUM ACCUMULATION IN THE PROXIMAL CONVOLUTED TUBULES OF THE FROG'S KIDNEY

1. In a manner similar to that of the sartorius muscle, the isolated kidney of the frog can accumulate K against a gradient to upwards of three times its normal concentration. 2. The K-accumulating region is identified as the proximal tubule, which in the isolated tissue immersed over 24 hours in the cold (2–3°C.) amounts to about 90 per cent of the nephron minus the glomerulus. In the fresh tissue it constitutes about 70 per cent. The cells of the proximal tubule are impermeable to Na, but freely permeable to K and Cl. 3. The distal tubule in the isolated kidney does not accumulate K over the external concentration. The cells are permeable to Na which they actively extrude. This extrusion of Na goes parallel with a loss of osmotically associated water amounting to about 15 per cent of the weight of the fresh kidney, but varying somewhat with the conditions. 4. The accumulation of K in the proximal tubules is in accordance with the equations established for the sartorius muscle, and, as theoretically expected, there is no volume increase (but rather a small decrease) with the large accumulations, when the external Na concentration is maintained throughout. 5. With K accumulation in isotonic mixtures large volume changes occur as K is progressively substituted for Na. Over the range of external K concentration of 10 to 100 mM per litre the weight of the whole kidney changes to 2.5 times and the water of the cells of the proximal tubules increases to over four times. Up to an external K value of 90 mM per litre the mean weight of the kidney shows a linear relation when plotted against the reciprocal of the Na concentration plus the small glucose and Ca concentration. This relation is interpreted theoretically. 6. The effect of cyanide in the isotonic mixtures is to prevent the contraction of the distal tubules and to cause swelling of the same. It does not affect the volume, volume changes, or differential permeability of the proximal tubule. At the same time the membranes of the proximal tubule cells lose their characteristic permeability at a lower level of distension in the presence of cyanide. 7. The mean Na ratio for the kidney after 24 hours'' immersion in the cold is 0.26 ± 0.014 (giving standard deviation of mean). The ratio is defined as See PDF for Equation. For the fresh kidney the mean ratio is 0.39 ± 0.006. 8. The mean inulin ratio (28 observed in the cold) is 0.23 ± 0.012 and the same value for 10 observed at room temperature. At room temperature—2 hour immersion—the ratio is increased by cyanide to a mean of 0.32 ± 0.028, but only a slight increase is caused by cyanide in the cold. 9. The mean hemoglobin ratio after 24 hours'' immersion in the cold is 0.17 ± 0.004 and is unaffected by cyanide.     

Quantitative trait loci for foliar terpenes in a global eucalypt species

Terpenes are a diverse group of plant secondary metabolites that mediate a plethora of ecological interactions in many plant species. Despite increasing research into the genetic control of important adaptive traits in some plant species, the genetic control of terpenes in forest tree species is still relatively poorly studied. In this study, we use quantitative genetic and quantitative trait loci (QTL) analysis to investigate the genetic control of foliar terpenes in an ecologically and commercially important eucalypt species, Eucalyptus globulus. We show a moderate to high within-family broad-sense heritability and significant genetic basis to the variation in 14 of the 16 terpenes assayed. This is the first report of QTL for terpenes in this species. Eleven QTL influenced the terpenes overall. One QTL on linkage group 6 affected six of the seven different sesquiterpenes assayed (plus one monoterpene), which, in combination with highly significant correlations between these compounds, argues that their variation is influenced by a QTL with pleiotropic effect early in the biosynthetic pathway. We examine the homology of these QTL to those found in a closely related eucalypt, Eucalyptus nitens, and provide evidence that both common and unique QTL influence terpene levels.     

Ketonaemia in Uncontrolled Diabetes Mellitus

Blood ketone bodies, serum insulin levels, and plasma free fatty acids were examined in a series of patients with “non-ketotic diabetic coma” and compared with the findings in ketoacidotic subjects. Serum insulin levels in six “non ketotic” patients ranged between 1 and 25 μu./ml. and were not significantly different from levels reported in patients with ketoacidosis. In addition, plasma free fatty acids were shown to be unrelated to the degree of ketonaemia. The investigation shows that neither the levels of serum insulin nor those of free fatty acids can explain the absence of hyperketonaemia in some cases.     

Early vigour in wheat: Could it lead to more severe terminal drought stress under elevated atmospheric [CO2] and semi‐arid conditions?

Early vigour in wheat is a trait that has received attention for its benefits reducing evaporation from the soil surface early in the season. However, with the growth enhancement common to crops grown under elevated atmospheric CO₂ concentrations (e[CO₂]), there is a risk that too much early growth might deplete soil water and lead to more severe terminal drought stress in environments where production relies on stored soil water content. If this is the case, the incorporation of such a trait in wheat breeding programmes might have unintended negative consequences in the future, especially in dry years. We used selected data from cultivars with proven expression of high and low early vigour from the Australian Grains Free Air CO₂ Enrichment (AGFACE) facility, and complemented this analysis with simulation results from two crop growth models which differ in the modelling of leaf area development and crop water use. Grain yield responses to e[CO₂] were lower in the high early vigour group compared to the low early vigour group, and although these differences were not significant, they were corroborated by simulation model results. However, the simulated lower response with high early vigour lines was not caused by an earlier or greater depletion of soil water under e[CO₂] and the mechanisms responsible appear to be related to an earlier saturation of the radiation intercepted. Whether this is the case in the field needs to be further investigated. In addition, there was some evidence that the timing of the drought stress during crop growth influenced the effect of e[CO₂] regardless of the early vigour trait. There is a need for FACE investigations of the value of traits for drought adaptation to be conducted under more severe drought conditions and variable timing of drought stress, a risky but necessary endeavour.     

Unhealthy Days and Quality of Life in Irish Patients with Diabetes

Objectives

To study the determinants of health-related quality of life (HRQoL) in Irish patients with diabetes using the Centres for Disease Controls'' (CDC''s) ‘Unhealthy Days’ summary measure and to assesses the agreement between this generic HRQoL measure and the disease-specific Audit of Diabetes Dependant Quality of Life (ADDQoL) measure.

Research Design and Methods

Data were analysed from the Diabetes Quality of Life Study, a cross-sectional study of 1,456 people with diabetes in Ireland (71% response rate). Unhealthy days were assessed using the CDC''s ‘Unhealthy days’ summary measure. Quality of life (QoL) was also assessed using the ADDQoL measure. Analyses were conducted primarily using logistic regression. The agreement between the two QoL instruments was measured using the kappa co-efficient.

Results

Participants reported a median of 2 unhealthy days per month. In multivariate analyses, female gender (P = 0.001), insulin use (P = 0.030), diabetes complications (P = <0.001) were significantly associated with more unhealthy days. Older patients had fewer unhealthy days per month (P = 0.003). Agreement between the two measures of QoL (unhealthy days measure and ADDQoL) was poor, Kappa = 0.234

Conclusions

The findings highlight the determinants of HRQoL in patients with diabetes using a generic HRQoL summary measure. The ‘Unhealthy Days’ and the ADDQoL have poor agreement, therefore the ‘Unhealthy Days’ summary measure may be assessing a different construct. Nonetheless, this study demonstrates that the generic ‘Unhealthy Days’ summary measure can be used to detect determinants of HRQoL in patients with diabetes.     

Neuromuscular Electrical Stimulation as a Method to Maximize the Beneficial Effects of Muscle Stem Cells Transplanted into Dystrophic Skeletal Muscle

Cellular therapy is a potential approach to improve the regenerative capacity of damaged or diseased skeletal muscle. However, its clinical use has often been limited by impaired donor cell survival, proliferation and differentiation following transplantation. Additionally, functional improvements after transplantation are all-too-often negligible. Because the host microenvironment plays an important role in the fate of transplanted cells, methods to modulate the microenvironment and guide donor cell behavior are warranted. The purpose of this study was to investigate whether the use of neuromuscular electrical stimulation (NMES) for 1 or 4 weeks following muscle-derived stem cell (MDSC) transplantation into dystrophic skeletal muscle can modulate the fate of donor cells and enhance their contribution to muscle regeneration and functional improvements. Animals submitted to 4 weeks of NMES after transplantation demonstrated a 2-fold increase in the number of dystrophin+ myofibers as compared to control transplanted muscles. These findings were concomitant with an increased vascularity in the MDSC+NMES group when compared to non-stimulated counterparts. Additionally, animals subjected to NMES (with or without MDSC transplantation) presented an increased maximal specific tetanic force when compared to controls. Although cell transplantation and/or the use of NMES resulted in no changes in fatigue resistance, the combination of both MDSC transplantation and NMES resulted in a faster recovery from fatigue, when compared to non-injected and non-stimulated counterparts. We conclude that NMES is a viable method to improve MDSC engraftment, enhance dystrophic muscle strength, and, in combination with MDSC transplantation, improve recovery from fatigue. These findings suggest that NMES may be a clinically-relevant adjunct approach for cell transplantation into skeletal muscle.