Evidence for control of mast cell granule protease in situ by low pH

The second order rate constant, k2, for the inhibition of mast cell protease I by phenylmethanesulfonyl fluoride (PMSF) is lower for intact mast cells and isolated granules with intact membranes than for granules stripped of their membranes and suspended in medium at pH 7.1. In order to test the hypothesis that the decreased activity of the protease in intact granules is attributable to low pH, two agents capable of lowering pH in intracellular compartments similar to mast cell granules were tested. Ammonium chloride increased k2 of the protease in isolated granules with intact membranes and mast cells and wash out of the salt partially reversed this effect. Treatment of cells with nigericin also substantially increased the rate of protease inactivation by PMSF. These results are consistent with the proposal that the observed k2 is determined in whole or part by a low pH of the granule in situ or isolated with intact membranes. If the low k2 in situ is solely dependent on low pH, then the rate of protease inhibition can be utilized as an endogenous probe of granule pH. On this basis we have estimated the pH of the intracellular granule as 5.2 and that of the isolated granule with its membrane intact as 6.0. The value for the pH of granules in situ is lower than that previously estimated, and we have considered possible bases for this discrepancy.     

On the use of ordinal scoring scales in social life cycle assessment

The International Journal of Life Cycle Assessment -     

A non‐helical region in transmembrane helix 6 of hydrophobic amino acid transporter MhsT mediates substrate recognition

MhsT of Bacillus halodurans is a transporter of hydrophobic amino acids and a homologue of the eukaryotic SLC6 family of Na⁺‐dependent symporters for amino acids, neurotransmitters, osmolytes, or creatine. The broad range of transported amino acids by MhsT prompted the investigation of the substrate recognition mechanism. Here, we report six new substrate‐bound structures of MhsT, which, in conjunction with functional studies, reveal how the flexibility of a Gly‐Met‐Gly (GMG) motif in the unwound region of transmembrane segment 6 (TM6) is central for the recognition of substrates of different size by tailoring the binding site shape and volume. MhsT mutants, harboring substitutions within the unwound GMG loop and substrate binding pocket that mimick the binding sites of eukaryotic SLC6A18/B0AT3 and SLC6A19/B0AT1 transporters of neutral amino acids, exhibited impaired transport of aromatic amino acids that require a large binding site volume. Conservation of a general (G/A/C)ΦG motif among eukaryotic members of SLC6 family suggests a role for this loop in a common mechanism for substrate recognition and translocation by SLC6 transporters of broad substrate specificity.     

Offspring are lighter at birth and smaller in adulthood when born after a brother versus a sister in humans

In mammals, including humans, it is more costly to produce sons than it is to produce daughters, with maternal survival and subsequent reproductive success diminished more by producing male over female offspring. It is therefore predicted that offspring who are produced by mothers who have previously produced sons versus daughters will be compromised by the relatively high cost their mother incurred in the previous reproductive episode. Such effects are potentially important because characters that determine offspring survival and fecundity ultimately contribute to maternal fitness. Using questionnaire-based data from a contemporary human population, I show that birthweight (irrespective of their sex) is lower in individuals born after an elder brother than in those born after an elder sister. In addition, I show that both men and women who were born after a male versus a female sibling have reduced adulthood height, a known correlate of reproductive success in both sexes. The results suggest that producing sons may have a negative effect on the fitness of subsequent offspring, which has implications for calculations of maternal fitness and for optimal sex allocation.     

The location of Taenia pisiformis, Taenia ovis and Taenia hydatigena in the gut of the dog and its effect on net environmental contamination with ova.

Autopsy of dogs 56 days after infection with either T. pisiformis, T. ovis or T. hydatigena showed that these worms could be found attached at any point along the length of the small intestine, but were most commonly in the anterior half. The mean relaxed lengths of T. pisiformis, T. ovis and T. hydatigena were 107 cm, 156 cm and 177 cm respectively. Attached gravid proglottides contained a mean of 41 000 eggs each in T. pisiformis, 31 000 eggs in T. hydatigena and 95000 eggs in T. ovis, whereas proglottides free in the gut contained means of only 1370, 500 and 1400 eggs respectively; therefore, the majority of eggs were released into the gut before segments passed out into the faeces. It was shown that eggs of all 3 species of worms hatched and activated in the small intestine of the dog, especially in the anterior half. Eggs of T. pisiformis which had been passaged through the intestine of the dog and stored in the faeces for 5 days were poorly infective for rabbits compared with eggs only stored in faeces. It was concluded, therefore, that during taeniid infections of dogs the point of apolysis in the gut plays a significant role in determining environmental contamination with eggs. Puppies which had been fed 10000 T. ovis eggs daily for 6 weeks prior to infection with T. ovis cysticerci showed no difference in susceptibility to the infection when compared with untreated puppies.