Evidence for a single glutamate receptor of the ionotropic kainate/quisqualate type

The kainate (KA) and the quisqualate (QUIS) receptors that activate cation channels in the central nervous system have previously been defined as two of the major glutamate receptor types. In amphibian brain, an exceptionally rich source of these sites, they can be coextracted by octylglucoside and shown to behave as one entity in all analyses made in solution. When partly purified by lectin affinity, ion-exchange chromatography or by sucrose gradient centrifugation, the two activities comigrate in a 1:1 ratio. When the QUIS component is bound to an immobilized specific QUIS agonist, the KA component is extracted in parallel with it. There are equivalent numbers of the QUIS and KA sites and the two sites show a single affinity series for the binding of glutamatergic agonists. We deduce that KA or QUIS select different conformations of a single KA/QUIS receptor binding site, leading thus to the different channel-opening events that have been reported for these two agonists.     

Effects of peptidoglycans from periodontal pathogens on selected biological activities of CD-1 murine peritoneal macrophages

Resident CD-1 murine peritoneal macrophages were exposed to various concentrations of purified peptidoglycan isolated from members of the genera Bacteroides, Eikenella, and Actinomyces. Macrophage viability, the release of lysozyme, acid phosphatase, and prostaglandins E1 and E2 were assayed as a function of peptidoglycan concentration and time. Macrophages responded as a function of peptidoglycan concentration with increased release of acid phosphatase and prostaglandins; all cells remained greater than 90% viable during the course of the experiments. However, concentrations of peptidoglycan greater than 50 micrograms/mL were toxic to the macrophages, while the peptidoglycan from B. capillus strain 925.08 and Actinomyces viscosus strain T14AV consumed complement by both the classical and the alternate pathways. Cellular lysozyme activity and phagocytosis of Saccharomyces cerevisiae were significantly reduced in the presence of peptidoglycan. When viewed by scanning electron microscopy, the activated macrophages were rounded, lacked distinct pseudopod extensions, and possessed an increased number of microvilli and plasma membrane associated vesicles. These morphological alterations occurred as early as 3 h. Transmission electron microscopy revealed the purified peptidoglycan to have been taken up into numerous phagosomes; however, even after 24 h incubation, it was only partially degraded.     

Purification of the neurotensin receptor from bovine brain

The neurotensin receptor protein, solubilized with digitonin/asolectin from bovine cerebral cortex membranes, was purified to apparent homogeneity by affinity chromatography using immobilized neurotensin. The product exhibits saturable and specific binding of [3,11-tyrosyl-3,5-3H]neurotensin with an apparent affinity (Kd = 5.5 nM) comparable to that measured in intact membranes and crude soluble extracts. The affinity-purified material, after reduction with 100 mM dithiothreitol, in denaturing gel electrophoresis showed a single polypeptide of Mr 72,000. Under nonreducing conditions the apparent Mr, however, was 50,000, suggesting the presence of intramolecular disulfide bonds. The purified neurotensin receptor was judged to be homogeneous, in that (i) only a single polypeptide was detectable; and (ii) the overall purification was 30,000-50,000-fold, giving a specific neurotensin-binding activity close to the theoretical maximum.     

Accumulating wing damage affects foraging decisions in honeybees (Apis mellifera L.)

Abstract.  1. Nectar-foraging honeybees ( Apis mellifera ) on lavender ( Lavandula stoechas ) appear to forage so as to maximise net energy return from foraging bouts; however, evidence from other studies suggests that foraging has a detrimental effect on survival, due at least in part to physiological deterioration of the flight mechanism. But foragers also acquire wing damage during foraging, which may increase foraging effort and reduce foraging lifespan.
2. The accumulation of damage over time and its effects on foraging flight and flower choice were studied in the field using a system in which the criteria for flower preference by foragers was known from previous work. Wing damage accumulated exponentially over time and resulted in foragers becoming less choosy about the flowers they visited.
3. Damage added experimentally contributed independently to the effect on choosiness. Effects of wing damage (natural and added experimentally) were also independent of those of a relative measure of age, which related in an inconsistent way to changes in foraging preferences.     

Female streetworking prostitution and HIV infection in Glasgow.

OBJECTIVES--To identify the extent of HIV infection and injecting drug use among female streetworking prostitutes in Glasgow; to estimate the size of the female streetworking prostitute population in the city; and to estimate the number of HIV positive women working as prostitutes on the streets in Glasgow. DESIGN--Observation and interviewing of female prostitutes over seven months in red light district; analysis of saliva samples for presence of antibodies to HIV; capture-recapture approach to estimating the size of the female streetworking prostitute population. SETTING--Glasgow. SUBJECTS--206 female streetworking prostitutes. MAIN OUTCOME MEASURES--Number of women with antibodies to HIV, self reported use of injecting drugs, history of contact with 206 women. RESULTS--Saliva samples were requested from 197 women; 159 (81%) provided samples. Four (2.5%, 95% confidence interval 0.7%-6.3%) of the samples were positive for HIV, all of which had been provided by women who injected drugs. Of the 206 streetworking women contacted 147 (71%) were injecting drug users. About 1150 women are estimated to work on the streets in Glasgow over a 12 month period. CONCLUSIONS--HIV is not as widespread among female prostitutes as many reports in the tabloid press suggest. A greater proportion of female streetworking prostitutes in Glasgow are injecting drugs than has been reported for other British cities.     

The poly-beta-hydroxybutyrate granule in vivo. A new insight based on NMR spectroscopy of whole cells

High resolution 13C NMR spectroscopy of live cells has been used to show that poly-beta-hydroxybutyrate (PHB) is predominantly in a mobile state within the storage granules of Alcaligenes eutrophus, Methylobacterium extorquens, and Methylobacterium AM1. Comparison of chemical and NMR analysis of PHB indicates that about 70% of the polymer in A. eutrophus gives sharp observable resonances. Temperature-dependent line widths and relaxation rates together with nuclear Overhauser effect measurements demonstrate that the observed material is effectively a mobile amorphous elastomer that is well above its glass transition temperature. The hydroxyvalerate-hydroxybutyrate copolymer produced by propionate-fed A. eutrophus has virtually the same mobility as the homopolymer. Evidence is presented indicating that water is an integral component of the PHB granule and that this component acts as a plasticizer for the polymer. These observations strongly suggest that the enzyme(s) responsible for PHB biosynthesis and consumption operate only on mobile hydrated material and that the solid granules characteristic of dried cells are partially artifactual. This model is supported by a reinterpretation of previously inexplicable biochemical results.     

The Interaction of Drugs with DNA Gyrase: A Model for the Molecular Basis of Quinolone Action

Abstract

DNA gyrase supercoils DNA in bacteria. The fact that it is essential in all bacteria and absent from eukaryotes makes it an ideal drug target. We discuss the action of coumarin and quinolone drugs on gyrase. In the case of coumarins, the drugs are known to be competitive inhibitors of the gyrase ATPase reaction. From a combination of structural and biochemical studies, the molecular details of the gyrase-coumarin complex are well established. In the case of quinolones, the drugs are thought to act by stabilising a cleavage complex between gyrase and DNA that arrests polymerases in vivo. The exact nature of the gyrase-quinolone-DNA complex is not known; we propose a model for this complex based on structural and biochemical data.