Effect of elevated temperature on osmotic and ionic regulation in a salt-tolerant caddisfly from Death Valley,California

The effects of temperature (8–10 or 20°C) on regulation of haemolymph osmotic and ionic concentrations were investigated over a range of salinities (0–25‰) in fifth-instar larvae of the Death Valley caddisfly Limnephilus assimilis. At low temperatures, levels of chloride and sodium in the haemolymph are regulated over a wide range of salinities corresponding to the salinities at which larvae occur in nature and at which they can complete development into adults. In contrast, haemolymph osmolality is constant at low salinities (<14‰) but approaches conformity with the medium at higher salinities. High temperature reduces the larva's ability to maintain low chloride concentrations in its haemolymph and also leads to a reduction in haemolymph osmotic pressure; thus, at high temperatures ions account for more of the haemolymph osmotic concentration than at low temperatures. These data suggest that the absence of larvae from thermal pools and from all Death Valley waters in summer can be explained by the effects of high water temperatures on hydromineral regulation.     

Volatile chemical release by bethylid wasps: identity, phylogeny, anatomy and behaviour

The structures of volatile chemicals released by parasitic wasps in the family Bethylidae are shown to correspond to the subfamily to which the species belong. Species in the Epyrinae release skatole (3-methylindole) and species in the Bethylinae release a spiroacetal (2-methyl-1,7-dioxaspiro [5.5]undecane): these compounds are chemically very different. The enantiomeric composition of the spiroacetal differs between congeneric species. Chemical release is a discrete event under the active control of both male and female wasps. Structural differences between the mandibular glands and intramandibular glands suggest the mandibular glands to be the source of released volatiles. Real-time mass spectrometry shows that the spiroacetal is released by Goniozus nephantidis females during dyadic resource contests, with release more common during more aggressive interactions. Chemical tagging with deuterium further shows that the volatile is released by the loser of an agonistic interaction and not the winner. The function of spiroacetal and skatole release by bethylids is discussed.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 837–852.     

The abundance of phytophilous invertebrates on different species of submerged macrophytes*

SUMMARY. 1. We tested Krecker's model (1939) which states that the abundance of invertebrates per unit macrophyte biomass varies with plant species and is higher on plants with finely dissected leaves than on plants with broad leaves. The abundance of invertebrates was measured in thirteen lacustrine macrophyte beds in southern Québec, Canada. The model was tested for the total abundance of invertebrates and for the abundances of Chironomidae, Cladocera, Cyclopoida, Gastropoda, Hydracarina, Ostracoda and Trichoptera. 2. More epiphytic invertebrates were found on the dissected Myriophyllum spp. than on the broad-leaved Potamogeton amplifolius Tuckerm, P. robbinsii Oakes and Vallisneria americana Michx. (P<0.01). More invertebrates were also found on P. amplifolius than on P. robbinsii or V. americana (P<0.01). The total abundance of invertebrates was not systematically related to the degree of plant dissection. 3. The abundances of Chironomidae, Cladocera, Cyclopoida, Gastropoda, Hydracarina, Ostracoda and Trichoptera varied on different plant species (P<0.01). Contrary to Krecker's hypothesis, however, macrophytes with finely dissected leaves (Ceratophyllum demersum and Myriophyllum spp.) did not in general support more invertebrates per unit plant biomass than plants with large leaves (Potamogeton amplifolius, P. robbinsii and Vallisneria americana).     

Enhanced inhibition of tissue factor by the extended form of an endothelial cell glycoprotein (an extrinsic pathway inhibitor)

We have identified, in the supernatant medium of cultured endothelial cells, an additional inhibitor of tissue factor that is eluted at higher salt concentrations during heparin-Sepharose chromatography and is a much more potent inhibitor of the activation of the coagulation cascade than the species we studied earlier (Colburn and Buonassisi: In Vitro Cellular and Developmental Biology 24: 1133-1136, 1988). The inhibitor we described earlier has been shown to be functionally and immunologically identical to a rabbit plasma extrinsic pathway inhibitor, EPI (Warn-Cramer et al.: Thrombosis Research 61:515-527, 1991). The new species (molecular mass, 47 kDa) is susceptible to proteolytic cleavage which results in a sharp reduction of its ability to inhibit tissue factor and an increase in electrophoretic mobility compatible with a molecular mass of 45 kDa, the same as that of the inhibitor reported earlier. Based on these data, we suggest that the new inhibitor yields, through proteolytic cleavage of an amino acid sequence of about 25 residues, the N-glycan-sulfated compound previously described.     

Listeria species in a California coast estuarine environment

Listeria species and L. monocytogenes were found in 81 and 62%, respectively, of fresh or low-salinity waters (37 samples) in tributaries draining into Humboldt-Arcata Bay, Calif., during a winter (January-February) sampling period. The incidence of Listeria species and L. monocytogenes in sediment (46 samples) from the same sites where water was sampled was 30.4 and 17.4%, respectively. One of three bay water samples contained Listeria species (including L. monocytogenes), while of 35 samples of oysters examined, only 1 was found positive for Listeria species (L. innocua). A given species or L. monocytogenes serogroup appeared to predominate in fresh water when domesticated animals (cows, horses) were nearby, whereas greater variety with no species predominance was observed in areas with no direct animal influence.     

Bayesian-integrated microbial forensics

In the aftermath of the 2001 anthrax letters, researchers have been exploring ways to predict the production environment of unknown-source microorganisms. Culture medium, presence of agar, culturing temperature, and drying method are just some of the broad spectrum of characteristics an investigator might like to infer. The effects of many of these factors on microorganisms are not well understood, but the complex way in which microbes interact with their environments suggests that numerous analytical techniques measuring different properties will eventually be needed for complete characterization. In this work, we present a Bayesian statistical framework for integrating disparate analytical measurements. We illustrate its application to the problem of characterizing the culture medium of Bacillus spores using three different mass spectral techniques. The results of our study suggest that integrating data in this way significantly improves the accuracy and robustness of the analyses.     

A semiempirical model for the electrophoretic mobilities of peptides in free-solution capillary electrophoresis

In this study an attempt is made to explore the effect of a peptide's size, charge, and hydrophobicity on its electrophoretic mobility (mu) as measured by free-solution capillary electrophoresis with the aim of developing a semiempirical model which incorporates these effects. The effects of peptide size (which is measured by the number of amino acids in the polypeptide chain (n] and charge on mu are independently determined by experiment in a single solvent system and combined to give the relationship (formula; see text) where the constant 5.23 X 10(-4) is postulated to depend on the solvent system used. The form of Eq. [A.1] was confirmed, and the values of the constants 5.23 X 10(-4) and 2.47 X 10(-5) were determined, by measuring the electrophoretic mobilities of 40 peptides varying in size from 3 to 39 amino acids and varying in charge from 0.33 to 14.0. Furthermore, the effect of noncharged neutral amino acids on mobility was investigated and shown to be present, but only as a minor perturbation on the effects of size and charge.     

Tumor promoter benzoyl peroxide induces sulfhydryl oxidation in protein kinase C: its reversibility is related to the cellular resistance to peroxide-induced cytotoxicity

Since tumor promoter benzoyl peroxide (BPO) mimics phorbol esters in some aspects, its effects on protein kinase C (PKC) were previously studied. However, in those studies due to the presence of thiol agents in the PKC preparations, the sensitive reaction of BPO with redox-active cysteine residues in PKC was not observed. In this study, by excluding thiol agents present in the purified PKC preparation, low concentrations of BPO modified PKC, resulting in the loss of both kinase activity and phorbol ester binding (IC50 = 0. 2 to 0.5 microM). This modification, which was not dependent on transition metals, was totally blocked by a variety of thiol agents including GSH, which directly reacted with BPO. Substoichiometric amounts of BPO (0.4 mol/mol of PKC) oxidized two sulfhydryls in PKC and inactivated the enzyme which was readily reversed by dithiothreitol. The regulatory domain having zinc thiolate structures supporting the membrane-inserting region provided the specificity for PKC reaction with BPO, which partitioned into the membrane. Unlike H2O2, BPO did not induce the generation of the Ca2+/lipid-independent activated form of PKC. Other redox-sensitive enzymes such as protein kinase A, phosphorylase kinase, and protein phosphatase 2A required nearly 25- to 100-fold higher concentrations of BPO for inactivation. BPO also inactivated PKC in a variety of cell types. In the JB6 (30 P-) nonpromotable cell line and other normal cell lines, where BPO was more cytotoxic, it readily inactivated PKC due to a slow reversibility of this inactivation by the cell. However, in the JB6 (41 P+) promotable cell line, C3H10T1/2 and B16 melanoma cells, where BPO was less cytotoxic, it did not readily inactivate PKC due to a rapid reversibility of this inactivation by an endogenous mechanism. Nevertheless, BPO inactivated PKC at an equal rate in the homogenates prepared from all these cell types. Inclusion of NADPH reversed this inactivation in the homogenates to a different extent, presumably due to a difference in distribution of a protein disulfide reductase, which reverses this oxidative modification. BPO-induced modification of PKC occurred independent of the cellular status of GSH. However, externally added GSH and cell-impermeable thiol agents prevented the BPO-induced modification of PKC. Since BPO readily partitions into membranes, its reaction with redox-cycling thiols of membrane proteins such as PKC may trigger epigenetic events to prevent cytotoxicity, but favor tumor promotion.     

Sites phosphorylated in myosin light chain in contracting smooth muscle

Purified smooth muscle myosin light chain can be phosphorylated at multiple sites by myosin light chain kinase and protein kinase C. We have determined the sites phosphorylated on myosin light chain in intact bovine tracheal smooth muscle. Stimulation with 10 microM carbachol resulted in 66 +/- 5% monophosphorylated and 11 +/- 2% diphosphorylated myosin light chain after 1 min, and 47 +/- 4% monophosphorylated and 5 +/- 2% diphosphorylated myosin light chain after 30 min. Myosin heavy chain contained 0.06 +/- 0.01 mol of phosphate/mol of protein which did not change with carbachol. At both 1 and 30 min the monophosphorylated myosin light chain contained only phosphoserine whereas the diphosphorylated myosin light chain contained both phosphoserine and phosphothreonine. Two-dimensional peptide mapping of tryptic digests of monophosphorylated and diphosphorylated myosin light chain obtained from carbachol-stimulated tissue was similar to the peptide maps of purified light chain monophosphorylated and diphosphorylated, respectively, by myosin light chain kinase; these maps were distinct from the map obtained with tracheal light chain phosphorylated by protein kinase C. Phosphorylation of tracheal smooth muscle myosin light chain by myosin light chain kinase yields the tryptic phosphopeptide ATSNVFAMFDQSQIQEFK with S the phosphoserine in the monophosphorylated myosin light chain and TS the phosphotreonine and phosphoserine in the diphosphorylated myosin light chain. Thus, stimulation of tracheal smooth muscle with a high concentration of carbachol results in formation of both monophosphorylated and diphosphorylated myosin light chain although the amount of diphosphorylated light chain is substantially less than monophosphorylated light chain. In the intact muscle, myosin light chain is phosphorylated at sites corresponding to myosin light chain kinase phosphorylation.