Physical map of the coliphage 186 chromosome

Summary A physical map of coliphage 186 was established by cloning various restriction fragments of 186 DNA into the plasmid vector pBR322 and determining the genes encoded on each fragment by marker rescue experiments using a range of 186 mutant phage.     

Phospholipase D in homogenates from HL-60 granulocytes: implications of calcium and G protein control

Occupancy of chemotactic peptide receptors leads to rapid initiation of phospholipase D (PLD) activity in intact dimethylsulfoxide-differentiated HL-60 granulocytes (Pai, J.-K, Siegel, M.I., Egan, R.W., and Billah, M.M. (1988) J. Biol. Chem. 263, 12472). To gain further insight into the activation mechanisms, PLD has been studied in cell lysates from HL-60 granulocytes, using 1-0-alkyl-2-oleoyl-[32P]phosphatidylcholine (alkyl-[32P]PC), 1-0-[3H]alkyl-2-oleoyl-phosphatidylcholine [( 3H]alkyl-PC) and [14C]arachidonyl-phospholipids as substrates. In the presence of Ca2+ and GTP gamma S, post-nuclear homogenates degrade alkyl-[32P]PC to produce 1-0-alkyl-[32P]phosphatidic acid (alkyl-[32P]-PA), and in the presence of ethanol, also 1-0-alkyl-[32P]phosphatidylethanol (alkyl-[32P]PEt). By comparing the 3H/32P ratios of PA and PEt to that of PC, it is concluded that PA and PEt are formed exclusively by a PLD that catalyzes both hydrolysis and transphosphatidylation between PC and ethanol. Furthermore, PC containing either ester- or ether-linkage at the sn-1 position is degraded in preference to phosphatidylethanolamine and phosphatidylinositol by PLD in HL-60 cell homogenates. It is concluded that HL-60 granulocytes contain a PC-specific PLD that requires both Ca2+ and GTP for activation.     

Comparison of hot water immersion at self-adjusted maximum tolerable temperature,with or without the addition of salt,for rapid weight loss in mixed martial arts athletes

Hot water immersion is used by athletes in weight category sports to produce rapid weight loss (RWL) by means of passive fluid loss, and often is performed with the addition of Epsom salts (magnesium sulphate). This study investigated the magnitude of body mass losses during hot water immersion with or without the addition of salt, with the temperature commencing at 37.8°C and being self-adjusted by participants to their maximum tolerable temperature. In a crossover design, eight male MMA athletes (29.4 ± 5.3 y; 1.83 ± 0.05 m; 85.0 ± 4.9 kg) performed a 20 min whole-body immersion followed by a 40 min wrap in a warm room, twice in sequence per visit. During one visit, only fresh water was used (FWB), and in the other visit, magnesium sulphate (1.6% wt/vol) was added to the bath (SWB). Prior to each visit, 24 h of carbohydrate, fibre and fluid restriction was undertaken. Water temperatures at the end of the first and second baths were ~39.0°C and ~39.5°C, respectively. Body mass losses induced by the hot bath protocols were 1.71 ± 0.70 kg and 1.66 ± 0.78 kg for FWB and SWB, respectively (P = 0.867 between trials, d = 0.07), and equivalent to ~2.0% body mass. Body mass lost during the entire RWL protocol was 4.5 ± 0.7%. Under the conditions employed, the magnitude of body mass lost in SWB was similar to FWB. Augmenting passive fluid loss during hot water immersion with the addition of salt may require a higher salt concentration than that presently utilised.     

Polarographic assay for phospholipase A2

A rapid, specific, and quantitative assay for phospholipase A₂ from Naja naja venom has been devised, in which phospholipid hydrolysis is measured as soybean lipoxidase-catalyzed oxygen incorporation into the ensuing unsaturated fatty acids. Under conditions where phospholipid was limiting, a linear relationship developed between the extent of oxygen uptake and the amount of egg lecithin metabolized. When phospholipase was rate limiting, the initial rate of oxygen consumption was a linear function of phospholipase concentration over a 14-fold range from 30 to 420 ng/ml. This linear relationship did not exist at higher phospholipase levels, probably due to the micellar nature of the phospholipid. Since this assay can readily detect as little as 17 ng/ml of phospholipase A₂ (Naja naja venom) and is insensitive to most potential interfering materials, it should be useful in a variety of applications.     

Genetic map of coliphage 186 from a novel use of marker rescue frequencies

Summary A genetic map of phage 186 has been constructed, using the frequency of marker rescue from 186 mutant prophages for genes to the left of att, and int promoted recombination for genes to its right. At the left end of the genome lie 7 genes involved in the formation of the phage head, followed to the right by the lysis gene P, a gene (O) of unknown function, and a group of 11 genes involved in the formation of the phage tail. Gene B, the late control gene, lies to the right of this group but to the left of the phage attachment site. To the right of the att site lie the non-essential genes (cI and cII) involved in lysogen formation and the gene (A) required for 186 DNA synthesis.