Calcium promotes cell survival through CaM-K kinase activation of the protein-kinase-B pathway

The protection against apoptosis provided by growth factors in several cell lines is due to stimulation of the phosphatidylinositol-3-OH kinase (PI(3)K) pathway, which results in activation of protein kinase B (PKB; also known as c-Akt and Rac) and phosphorylation and sequestration to protein 14-3-3 of the proapoptotic Bcl-2-family member BAD. A modest increase in intracellular Ca2+ concentration also promotes survival of some cultured neurons through a pathway that requires calmodulin but is independent of PI(3)K and the MAP kinases. Here we report that Ca2+/calmodulin-dependent protein kinase kinase (CaM-KK) activates PKB directly, resulting in phosphorylation of BAD on serine residue 136 and the interaction of BAD with protein 14-3-3. Serum withdrawal induced a three- to fourfold increase in cell death of NG108 neuroblastoma cells, and this apoptosis was largely blocked by increasing the intracellular Ca2+ concentration with NMDA (N-methyl-D-aspartate) or KCl or by transfection with constitutively active CaM-KK. The effect of NMDA on cell survival was blocked by transfection with dominant-negative forms of CaM-KK or PKB. These results identify a Ca2+-triggered signalling cascade in which CaM-KK activates PKB, which in turn phosphorylates BAD and protects cells from apoptosis.     

Lipid-lowering therapy for patients with or at risk of coronary artery disease

Several studies have shown that effective lipid-lowering therapy slows the progression of atherosclerotic lesions in the coronary and carotid arteries. Recent clinical trials have confirmed and extended previous work showing that lowering cholesterol reduces the risk of coronary events. A clear reduction in major coronary events during treatment for 5 years with the hydroxymethylglutaryl-coenzyme A reductase inhibitor pravastatin was observed in the West of Scotland study and in the preliminary results of the Cholesterol and Recurrent Events study. The Scandinavian Simvastatin Survival Study has provided the first unequivocal demonstration of improved survival as a result of lipid-lowering therapy. These three trials, which together included over 15,000 patients studied for 5 years, have provided good evidence that noncardiovascular mortality is not affected by substantial reductions in blood cholesterol.     

Gonadotropins are essential modifier factors for gonadal tumor development in inhibin-deficient mice

We previously demonstrated that mice deficient in inhibin develop gonadal sex cord-stromal tumors with nearly 100% penetrance. These ovarian and testicular tumors develop as early as 4 weeks of age and eventually cause cachexia-like symptoms and death in the inhibin-deficient mice. Gonadectomized inhibin-deficient mice initially do not develop this wasting syndrome, but eventually will develop adrenal cortical tumors with similar penetrance. These studies have demonstrated that inhibin is a secreted type of tumor suppressor in the gonads and adrenal glands. Gonadotropins are implicated to influence gonadal tumor development in humans as well as experimental animals, and in inhibin-deficient mice, serum FSH levels are elevated. To determine whether gonadotropins influence the development and/or progression of the tumors in the inhibin-deficient mice, we took advantage of a naturally occurring mutant mouse, hypogonadal (hpg); hpg/hpg mice lack a functional GnRH gene and, therefore, have suppressed FSH and LH levels. Heterozygous hpg/+mice were crossed to heterozygous inhibin mutant mice to generate compound homozygous mutant mice that lack both inhibin and GnRH. These compound homozygous mutant mice do not develop a wasting syndrome, do not exhibit gonadal or adrenal tumors, and can survive for more than 1 yr. These results demonstrate that gonadotropins are essential modifier factors for tumor development in inhibin-deficient mice.     

Who is driving when unrestrained children and teenagers are hurt?

This paper examines driver characteristics in crashes where child and teenage motor vehicle crash victims were injured, in particular factors that determine whether or not the victim was restrained. Analyzing the data on children and teenagers who are injured revealed that the presence of a second adult in a vehicle increased the likelihood that these passengers were unrestrained. Other findings are more predictable: victim restraint use generally mirrored driver restraint use; a male driver, a young driver, a drinking driver, a speeding or reckless driver, an unlicenced or suspended driver, and a night-time trip each independently raised the odds that child and teenage passengers were not restrained when they were injured.     

Empirical scoring functions. II. The testing of an empirical scoring function for the prediction of ligand-receptor binding affinities and the use of Bayesian regression to improve the quality of the model

This paper tests the performance of a simple empirical scoring function on a set of candidate designs produced by a de novo design package. The scoring function calculates approximate ligand-receptor binding affinities given a putative binding geometry. To our knowledge this is the first substantial test of an empirical scoring function of this type on a set of molecular designs which were then subsequently synthesised and assayed. The performance illustrates that the methods used to construct the scoring function and the reliance on plausible, yet potentially false, binding modes can lead to significant over-prediction of binding affinity in bad cases. This is anticipated on theoretical grounds and provides caveats on the reliance which can be placed when using the scoring function as a screen in the choice of molecular designs. To improve the predictability of the scoring function and to understand experimental results, it is important to perform subsequent Quantitative Structure-Activity Relationship (QSAR) studies. In this paper, Bayesian regression is performed to improve the predictability of the scoring function in the light of the assay results. Bayesian regression provides a rigorous mathematical framework for the incorporation of prior information, in this case information from the original training set, into a regression on the assay results of the candidate molecular designs. The results indicate that Bayesian regression is a useful and practical technique when relevant prior knowledge is available and that the constraints embodied in the prior information can be used to improve the robustness and accuracy of regression models. We believe this to be the first application of Bayesian regression to QSAR analysis in chemistry.     

Seasonal fluctuations in plasma concentrations of luteinizing hormone and progesterone in Brahman (Bos indicus) and Hereford-Shorthorn (Bos taurus) cows grazing pastures at two stocking rates in a subtropical environment

The effects of day length and grazing intensity on seasonal fluctuations in plasma concentrations of luteinizing hormone (LH) and cyclic ovarian activity were determined in Brahman (Bos indicus) and Hereford-Shorthorn (Bos taurus) cows maintained at two stocking rates in a subtropical environment. Contemporary groups of ovariectomised cows were monitored for fluctuations in plasma concentrations of LH. Equal numbers (n = 5) of entire and ovariectomised Brahman and Hereford-Shorthorn cows were assigned to a pasture with a greater or lesser stocking rate. Over a 15-month period, live weight was recorded weekly, and a blood sample was taken at the same time for measurement of plasma LH in entire and ovariectomised cows, and plasma progesterone in entire cows. Plasma concentrations of progesterone were used as an index of cyclic luteal function (time of cessation or onset of oestrous cycles). Regression coefficients were calculated to determine the least-order regression coefficient (LORC; range 1st to 10 order) for which time of year explained at least 50% (r2 > 0.05) of changes in live weight, plasma LH, or plasma progesterone; regression coefficients of 4th and 5th order indicated seasonally-related changes in these variables. For all cows, live weight was greatest in late summer to early autumn and lowest in winter. Changes in live weight were more closely related to seasonal changes in pasture availability for cows on pastures at a greater stocking rate (LORC 4th-5th) than for cows on pastures at a lesser stocking rate (LORC 1st-3rd). Cyclic ovarian activity ceased in four Hereford-Shorthorn cows on pastures at a greater stocking rate in late autumn to early winter, and onset of oestrous cycles did not occur in all of these cows until late spring. Oestrous cycles were not detected in one of five cows in the other groups during different periods of the study; however, there were no apparent patterns to cessation of oestrous cycles in these groups. There were no seasonally-related changes in plasma LH in entire cows, at either stocking rate (LORC 10th; r2 = 0.16 to 0.41). In contrast, distinct seasonal fluctuations in plasma LH occurred in ovariectomised cows, with increases in spring and winter. Environmental cues induced greater fluctuations in plasma LH in ovariectomised cows at a greater stocking rate (LORC 3rd-5th; r2 = 0.71 to 0.72) compared with ovariectomised cows at a lesser stocking rate (LORC 3rd-5th; r2 = 0.53 to 0.58). The findings demonstrated that marked seasonal changes in reproductive activity of the hypothalamic pituitary axis can occur in cows with B. indicus and B. taurus genotypes in a subtropical environment; however, changes in plasma concentrations of LH are only apparent in ovariectomised cows. Concentrations of plasma LH in ovariectomised Brahman and Hereford-Shorthorn cows increased during winter, when pasture availability was limiting and cyclic luteal function ceased in four of five Hereford-Shorthorn cows.     

Peak expiratory flow and the resistance of the mini-wright peak flow meter

The purpose of this study was to examine whether the resistance of the peak flow meter influences its recordings. One hundred and twelve subjects, (healthy nonsmokers and smokers and subjects with lung diseases) performed three or more peak expiratory flow (PEF) manoeuvres through a Fleisch pneumotachograph with and without a mini-Wright peak flow meter added in random order as a resistance in series. The results were as follows. In comparison with a pneumotachograph alone, peak flow measured with an added mini-Wright meter had a smaller within-test variation, defined as the difference between the highest and second highest values of PEF in a series of blows. The mean (SE) variation was 14 (1.3) L.min-1 and 19 (1.5) L.min-1 with and without meter added, respectively. In comparison with the pneumotachograph alone, the addition of the mini-Wright meter caused PEF to be underread, especially at high flows. The difference (PEF with meter minus PEF without meter) = -0.064 (average PEF) -8 L.min-1; R2 = 0.13. The mean difference was -7.8 (1.1) %, and increased numerically for a given PEF, when maximal expiratory flow when 75% forced vital capacity remains to be exhaled (MEF75%FVC) decreased. The reproducibility criteria for repeated measurements of peak flow are more appropriately set at 30 L.min-1 than the commonly used 20 L.min-1, because a within-test variation of less than 30 L.min-1 was achieved in 76% of the subjects without PEF meter inserted and in 88% with meter inserted, with no difference between healthy untrained subjects and patients. The resistance of the peak expiratory flow meter causes less variation in recordings but reduces peak expiratory flow, especially at high values and when the peak is large as compared with the rest of the maximal expiratory flow-volume curve.