No abstract

The physiologic responses to an intense heat acclimation (HA) regimen (treadmill, 41.2 degrees C, 8 days, 56 min exercise/44 min rest) and the effects on stress and fluid balance hormone responses were examined in 13 unacclimated male volunteers. Venous blood samples were collected before (PRE) and after (POST) exercise (days 1, 4, 8) and analyzed for plasma renin activity (PRA), aldosterone (ALD), cortisol (PC), plasma volume shifts (delta PV%), sodium concentration (Na+), and potassium concentration (K+). HA responses (day 1 vs day 8) indicated reduced strain (P less than 0.05): decreased heart rate, rectal temperature, skin temperature, improved defense of PV, and attenuated PC responses. While plasma Na+ demonstrated no change during daily exercise, K+ (P less than 0.01), PC, PRA, and ALD increased (P less than 0.05) more than delta PV%(day 1: -7.1%, day 8: -5.1%) accounted for. Na+ and K+ did not change as a result of HA, and there was no change in fluid balance hormones (e.g., PRA, ALD). It was concluded that this intense heat acclimation regimen reduced physiologic strain by mechanisms other than alterations in fluid balance hormones and offered few physiologic advantages which cannot be gained through conventional heat acclimation techniques (e.g., walking).

We have developed an introductory bioinformatics exercise for sophomore biology and biochemistry students that reinforces the understanding of the structure of a gene and the principles and events involved in its expression. In addition, the activity illustrates the severe effect mutations in a gene sequence can have on the protein product. Students search GenBank for the wild-type nucleotide sequence of the Caenorhabditis elegans unc-22 gene, the amino acid sequence of its gene product, and the nucleotide sequence of the transposon Tc5. The nucleotide sequences are manipulated using two programs in the Lasergene® software package from DNASTAR®. The first program, EditSeq®, enables students to experience the meticulous process required to precisely locate and remove intron sequences from the wild-type unc-22 allele to generate a cDNA sequence. The unc-22(r466) allele is generated by inserting the sequence of the transposon Tc5 into the appropriate location of the third exon in unc-22. The open reading frames of both cDNAs are located and then translated. MegAlign®, the second program, aligns the wild-type sequence of the UNC-22 protein and the wild-type and mutant protein sequences that were constructed. The degree of sequence similarity between the aligned proteins allows students to verify their success in processing the gene, as well as to visualize the truncated protein product from the Tc5 mutant allele. Student feedback and possible modifications to the exercise as well as supplemental exercises are also discussed.

A major function of insulin in target tissues is the activation of glycogen synthase. Phosphatidylinositol 3-kinase (PI3K) has been implicated in the insulin-induced activation of glycogen synthase, although the true function of this enzyme remains unclear. Data presented here demonstrate that the PI3K inhibitors wortmannin and LY-294002 block the insulin-stimulated activation of protein phosphatase 1 (PP1) in rat ventricular cardiomyocytes. This loss of phosphatase activation mimics that seen in diabetic cardiomyocytes, in which insulin stimulation fails to activate both PP1 and glycogen synthase. Interestingly, in diabetic cells, insulin stimulated PI3K activity to 300% of that in untreated controls, whereas this activity was increased by only 77% in normal cells. PI3K protein levels, however, were similar in normal and diabetic cells. Our results indicate that PI3K is involved in the stimulation of glycogen synthase activity by insulin through the regulation of PP1. The inability of insulin to stimulate phosphatase activity in diabetic cells, despite a significant increase in PI3K activity, suggests a defect in the insulin signaling pathway that contributes to the pathology of insulin-dependent diabetes.