Elemental composition of secretory granules in pancreatic islets of Langerhans.

We have characterized, by electron probe microanalysis, rapidly frozen cultured rat islets at the level of individual secretory granules. Elemental analysis of thin, dried cryosections showed that beta granules could be distinguished by high Zn, Ca, and S, whereas non-beta (mainly alpha) granules contained elevated P and Mg. Although a single granule type predominated in a particular cell, some rebel granules were found in A cells that had the compositional fingerprint of B cell granules. Zn, which was found in millimolar concentrations in B cell granules, was considered a marker for the insulin storage complex. The data indicate that non-B islet cells in the adult pancreas may produce insulin-containing organelles and that, when glucagon and insulin are coexpressed, these hormones are packaged in separate granules.     

Planktivory in benthic nototheniid fish in McMurdo Sound,Antarctica

Synopsis Four species of nototheniid fish were sampled from below the sea ice near Cape Armitage, McMurdo Sound:Pagothenia borchgrevinki from just below the ice 1.5 km offshore,Trematomus bernacchii, Trematomus hansoni andTrematomus centronotus from off the bottom in about 20 m of water near the shore. Scale worms and isopods were conspicuous non-planktonic prey, and present in the three benthic fish species. The planktonic pteropod molluscLimacina helicina was numerically common in all four species of fish. The planktonic hyperiid amphipodHyperiella dilatata was also found in all fish species. WhereasP. borchgrevinki is planktivorous in accord with its pelagic habit, theTrematomus spp. clearly also feed on plankton from the water column.T. hansoni is particularly planktivorous, taking smaller copepods thanP. borchgrevinki.     

A study of the mechanism of soil-phosphate uptake in relation to plant species

Summary The concept of plant available and unavailable soil phosphate has been examined by growing a range of plant species in soils well mixed with carrier free P³².It was found that although activities of 5–100 µC per kg soil caused changes in dry matter and P uptake, they had no influence on the specific activity per unit dose.Only small and agriculturally insignificant differences have been found in the proportion of soil P to added P³² taken up by the different species in the acid and neutral soils employed. It is considered that such differences as there are may be due to exchange of phosphate between seed and soil. Marked differences, however, occurred in the total phosphate absorbed by the crops.These results support the view that a fraction of the soil phosphate exists in a labile pool which will exchange with added P³² and will maintain a definite equilibrium potential in the soil solution. Plants do not appear to have a means of increasing significantly the size of the labile pool under the experimental conditions described. The disparity, however, between the total phosphate uptake especially in very deficient soils does suggest that certain species are more efficient than others in the absorption of soil phosphate at low potential.     

Internal pH crisis, lysine decarboxylase and the acid tolerance response of Salmonella typhimurium

Salmonella typhimurium possesses an adaptive response to acid that increases survival during exposure to extremely low pH values. The acid tolerance response (ATR) includes both log-phase and stationary-phase systems. The log-phase ATR appears to require two components for maximum acid tolerance, namely an inducible pH homeostasis system, and a series of acid-shock proteins. We have discovered one of what appears to be a series of inducible exigency pH homeostasis systems that contribute to acid tolerance in extreme acid environments. The low pH-inducible lysine decarboxylase was shown to contribute significantly to pH homeostasis in environments as low as pH 3.0. Under the conditions tested, both lysine decarboxylase and σ^s-dependent acid-shock proteins were required for acid tolerance but only lysine decarboxylase contributed to pH homeostasis. The cadBA operon encoding lysine decarboxylase and a lysine/cadaverine antiporter were cloned from S. typhimurium and were found to be 79% homologous to the cadBA operon from Escherichia coli . The results suggest that S. typhimurium has a variety of means of fulfilling the pH homeostasis requirement of the ATR in the form of inducible amino acid decarboxylases.     

Identification of alcaligin as the siderophore produced by Bordetella pertussis and B. bronchiseptica.

The siderophores produced by iron-starved Bordetella pertussis and B. bronchiseptica were purified and were found to be identical. Using mass spectrometry and proton nuclear magnetic resonance, we determined that the siderophore produced by these organisms was identical to alcaligin, a siderophore produced by Alcaligenes denitrificans.     

gamma-Carboxyglutamic acids 36 and 40 do not contribute to human factor IX function.

The gamma-carboxyglutamic acid (Gla) domains of the vitamin K-dependent blood coagulation proteins contain 10 highly conserved Gla residues within the first 33 residues, but factor IX is unique in possessing 2 additional Gla residues at positions 36 and 40. To determine their importance, factor IX species lacking these Gla residues were isolated from heterologously expressed human factor IX. Using ion-exchange chromatography, peptide mapping, mass spectrometry, and N-terminal sequencing, we have purified and identified two partially carboxylated recombinant factor IX species; factor IX/gamma 40E is uncarboxylated at residue 40 and factor IX/gamma 36,40E is uncarboxylated at both residues 36 and 40. These species were compared with the fully gamma-carboxylated recombinant factor IX, unfractionated recombinant factor IX, and plasma-derived factor IX. As monitored by anti-factor IX:Ca (II)-specific antibodies and by the quenching of intrinsic fluorescence, all these factor IX species underwent the Ca(II)-induced conformational transition required for phospholipid membrane binding and bound equivalently to phospholipid vesicles composed of phosphatidylserine, phosphatidylcholine, and phosphatidylethanolamine. Endothelial cell binding was also similar in all species, with half-maximal inhibition of the binding of 125I-labeled plasma-derived factor IX at concentrations of 2-6 nM. Functionally, factor IX/gamma 36,40E and factor IX/gamma 40E were similar to fully gamma-carboxylated recombinant factor IX and plasma-derived factor IX in their coagulant activity and in their ability to participate in the activation of factor X in the tenase complex both with synthetic phospholipid vesicles and activated platelets. However, Gla 36 and Gla 40 represent part of the epitope targeted by anti-factor IX:Mg(II)-specific antibodies because these antibodies bound factor IX preferentially to factor IX/gamma 36,40E and factor IX/gamma 40E. These results demonstrate that the gamma-carboxylation of glutamic acid residues 36 and 40 in human factor IX is not required for any function of factor IX examined.