Practical precooling: effect on cycling time trial performance in warm conditions.

The purpose of this study was to compare the effects of two practical precooling techniques (skin cooling vs. skin + core cooling) on cycling time trial performance in warm conditions. Six trained cyclists completed one maximal graded exercise test (VO2(peak) 71.4 +/- 3.2 ml x kg(-1) x min(-1)) and four approximately 40 min laboratory cycling time trials in a heat chamber (34.3 degrees C +/- 1.1 degrees C; 41.2% +/- 3.0% rh) using a fixed-power/variable-power format. Cyclists prepared for the time trial using three techniques administered in a randomised order prior to the warm-up: (1) no cooling (control), (2) cooling jacket for 40 min (jacket) or (3) 30-min water immersion followed by a cooling jacket application for 40 min (combined). Rectal temperature prior to the time trial was 37.8 degrees C +/- 0.1 degrees C in control, similar in jacket (37.8 degrees C +/- 0.3 degrees C) and lower in combined (37.1 degrees C +/- 0.2 degrees C, P < 0.01). Compared with the control trial, time trial performance was not different for jacket precooling (-16 +/- 36 s, -0.7%; P = 0.35) but was faster for combined precooling (-42 +/- 25 s, - .8%; P = 0.009). In conclusion, a practical combined precooling strategy that involves immersion in cool water followed by the use of a cooling jacket can produce decrease in rectal temperature that persist throughout a warm-up and improve laboratory cycling time trial performance in warm conditions.

Synthesis, characterization, and ab initio theoretical study of a molecularly imprinted polymer selective for biosensor materials.

Despite the complex phenomena involved in encoding template molecule information within stable synthetic polymers to yield selective and efficient molecular recognition processes, molecularly imprinted polymers (MIP) are increasingly finding broad areas of application. Molecular interactions, both during the polymerization of the functional monomers in the presence of the template and during the processes of specific recognition after template removal, are key determinants of an effective MIP. Covalent and noncovalent template imprinting have been employed to achieve specific recognition sites. In the present study, a molecularly imprinted biocompatible polymer, having a high capacity and affinity for the dye template, nickel(II) phthalocyanine tetrasulfonic acid, has been prepared. UV-visible spectroscopy, FTIR spectroscopy, and ICP analysis were used to investigate the aspects of the synthesis, binding capacity, and adsorption kinetics of the system. Poly(allylamine) cross-linked with epichlorohydrin has been used to represent an amino-functional receptor. Binding isotherms and capacities were correlated with the degree of template removal. Kinetic studies of binding allowed diffusion mechanisms to be evaluated for the fine particulate MIP. Ab initio molecular orbital calculations were performed using Hartree-Fock, MP2, and density functional theory methods to determine the most likely mechanisms of molecular imprinting. Suitable theoretical models have been constructed to mimic the interactions between the template molecule and the polymer. Simulation of the vibrational spectra was also undertaken to make meaningful assignments to experimentally determined spectral bands resulting from these template MIP receptor interactions.

Molecular mechanism of a COOH-terminal gating determinant in the ROMK channel revealed by a Bartter's disease mutation.

The ROMK subtypes of inward-rectifier K(+) channels mediate potassium secretion and regulate NaCl reabsorption in the kidney. Loss-of-function mutations in this pH-sensitive K(+) channel cause Bartter's disease, a familial salt wasting nephropathy. One disease-causing mutation truncates the extreme COOH-terminus and induces a closed gating conformation. Here we identify a region within the deleted domain that plays an important role in pH-dependent gating. The domain contains a structural element that functionally interacts with the pH sensor in the cytoplasmic NH(2)-terminus to set a physiological range of pH sensitivity. Removal of the domain shifts the pK(a) towards alkaline pH values, causing channel inactivation under physiological conditions. Suppressor mutations within the pH sensor rescued channel gating and trans addition of the cognate peptide restored pH sensitivity. A specific interdomain interaction was revealed in an in vitro protein-protein binding assay between the NH(2)- and COOH-terminal cytoplasmic domains expressed as bacterial fusion proteins. These results provide new insights into the molecular mechanisms underlying Kir channel regulation and channel gating defects that are associated with Bartter's disease.

Regulation of potassium channel Kir 1.1 (ROMK) abundance in the thick ascending limb of Henle's loop.

The renal outer medullary potassium channel (ROMK) of the thick ascending limb (TAL) is a critical component of the counter-current multiplication mechanism. In this study, two new antibodies raised to ROMK were used to investigate changes in the renal abundance of ROMK with treatments known to strongly promote TAL function. These antibodies specifically recognized protein of the predicted size of 45 kD in immunoblots of rat kidney or COS cells transfected with ROMK cDNA. Infusion of 1-deamino-(8-D-arginine)-vasopressin (dDAVP), a vasopressin V2 receptor-selective agonist, for 7 d into Brattleboro rats resulted in dramatic increases in apical membrane labeling of ROMK in the TAL of dDAVP-treated rats, as assessed by immunocytochemical analyses. Using immunoblotting, a more than threefold increase in immunoreactive ROMK levels was observed in the outer medulla after dDAVP infusion. Restriction of water intake to increase vasopressin levels also significantly increased TAL ROMK immunolabeling and abundance in immunoblots. In addition, dietary Na(+) levels were varied to determine whether ROMK abundance was also affected under other conditions known to alter TAL transport. Rats fed higher levels of sodium, as either NaCl or NaHCO(3) (8 mEq/250 g body wt per d), exhibited significantly increased density of the 45-kD band, compared with the respective control animals. Moreover, in rats fed a low-NaCl diet (0.25 mEq/250 g body wt per d), a 50% decrease in band density for the 45-kD band was observed (relative to control rats fed 2.75 mEq/250 g body wt per d of NaCl). These results demonstrate that long-term adaptive changes in ROMK abundance occur in the TAL with stimuli that enhance transport by this segment.