The ultrastructural localization of acid phosphatase,alkaline phosphatase and adenosine triphosphatase in induced goitres of Xenopus laevis Daudin tadpoles

Summary Tadpoles of Xenopus laevis were maintained in solutions of potassium perchlorate, potassium thiocyanate, thiourea and propyl-thiouracil (0.005% w/v). Metamorphosis was inhibited and within a short time large well-vascularized goitres developed. The localization of certain phosphatases at the ultrastructural level in these goitres is described and the possible roles played by these enzymes is discussed. Acid phosphatase was localized in follicle epithelial cells in dense droplets thought to be lysosomes or cytolysomes. Alkaline phosphatase and a Mg⁺⁺-activated ATP-ase were found on the outer surfaces of the membranes of follicular cells and on the walls of capillary blood vessels.This work was carried out during the tenure by one of us (R.C.) of a Medical Research Council Scholarship and forms part of a programme of research in amphibian thyroid physiology supported by a grant from the Medical Research Council, which we gratefully acknowledge.     

Food-deprivation-induced phase shifts in Sminthopsis macroura froggatti

Past research has shown that there is a circadian oscillator in laboratory rats that is entrained by restricted feeding schedules. However, in laboratory rats at least, the light-dark (LD) cycle is the dominant zeitgeber in the entrainment of wheel-running activity rhythms. Given that dasyurid marsupials are predominantly carnivorous, the episodic intake of food in the wild and the high nutritive content of that food suggest that food may be an important zeitgeber in these species. Twelve Sminthopsis macroura froggatti were presented with a daily meal at 0900 hr under an LD 12:12 cycle with lights-on at 0600 hr for 37 days. Activity in anticipation of the meal was observed in most animals. Following this, all animals were exposed to periods of 12-18 days ad lib. food interspersed with 3-day periods of deprivation--a technique used previously to demonstrate persistent meal-associated rhythms. The meal-associated activity rhythms previously observed in rats during the 3-day deprivation period were not seen, but the 3-day deprivation period produced large phase-shifts in the activity rhythms of several S.m. froggatti. It is concluded that meal feeding does not dominate the LD cycle in entraining dasyurid marsupials, but that the frequent observation of phase shifts suggests a different and, perhaps, stronger role for food intake in biological rhythmicity than has been observed previously in laboratory rats.     

Burrowing,grooming, and feeding behaviour of Paraceradocus,an antarctic amphipod genus (Crustacea)

Summary Species of the amphipod genus Paraceradocus found near the Antarctic Peninsula were observed in aquaria. The animals live under stones in burrows in sediment which they excavate with their gnathopods. During burrowing the animal regularly turns backwards in a somersault-like movement. Paraceradocus feeds on detritus, which is manipulated by the gnathopods and the antennes. The gnathopods are also used for grooming. A dense brush of setae at the medial surface of the carpi of the gnathopods I is used to clean off fine particles from the antennes. These particles are transferred to the maxillipeds and are ingested. The rear appendages are mainly cleaned by the gnathopods II. Young animals sitting between the gnathopods of the adult participate in feeding.     

Cloning and characterisation of the serC and aroA genes of Yersinia enterocolitica, and construction of an aroA mutant

A gene library of Yersinia enterocolitica 8081 was constructed in the cosmid vector pHC79. Recombinants containing the aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase, were identified by complementation of the aroA mutation in Escherichia coli K-12 strain AB2829. All six recombinant plasmids which complemented aroA also complemented the serC mutation in E. coli K-12 strain KL282. Tn5 mutagenesis suggested serC encoding 3-phosphoserine aminotransferase was the proximal gene in an operon with aroA. The nucleotide sequence of a 3-kb HindII-EcoRV fragment encoding the two genes was determined. The serC and aroA open reading frames contain 362 and 428 codons, respectively, and the deduced amino acid sequences share 78% and 81% homology, respectively, with the corresponding E. coli genes. Sequence inspection revealed no obvious terminators or promoters in the intergenic region. The cloned Y. enterocolitica aroA gene was inactivated in vitro and reintroduced into the parental Y. enterocolitica 8081 strain using the suicide vector pJM703.1. Stable aroA insertion mutants of Y. enterocolitica were isolated.     

The effect of glucose on the expression of extracellular protein genes by Staphylococcus aureus strain V8

The bacteria from overnight cultures (20 h) of S. aureus V8 and exp negative mutant K6812-1, grown, aerobically, in 3% (w/v) Tryptone Soya Broth, at 37 degrees C, were resuspended in fresh medium, in the case of the parent strain +/- 1% (w/v) glucose, without change in bacterial density. During a 6 h incubation period there was an approximate doubling of bacterial density, to the same level, in each case. However, exoprotein production by the mutant was only 20% that of the parent whilst the addition of glucose to the V8 strain resulted in a tenfold reduction in the exoprotein formed. SDS-polyacrylamide gel electrophoresis showed that the exoprotein patterns of both organisms after 6 h incubation were the same as those observed in the overnight cultures whilst the presence of 1% (w/v) extra glucose changed the pattern produced by the parent to one similar to that of the mutant. The results showed that conditions which lead to the rapid formation of glucose catabolites produced an effect consistent with the inhibition of the activity of the exp gene product.     

Effect of hypertension on elasticity and geometry of aortic tissue from dogs

Inflation-extension experiments were carried out on segments of the descending thoracic aortas from 4 normotensive and 4 hypertensive dogs rendered hypertensive using either unilateral or bilateral renal artery constriction. Intravascular pressures up to 200 mm Hg and axial forces up to 200 g were used. The external diameter of the segment and the distance between two longitudinally spaced gage marks were recorded photographically at each pressure-force level combination. Dimensions in the underformed configuration were measured at the end of the inflation-extension experiment. Data were analyzed for changes in geometry and force-deformation response. Results indicate that: 1. Under sustained hypertension the wall thickness in the underformed configuration increases with a concurrent reduction in the in-situ longitudinal extension ratio. 2. This dual tissue response accomplishes substantial reductions in the circumferential and longitudinal stresses from the levels that would be reached at equivalent pressures in the absence of these geometric changes. 3. At comparable intravascular pressures the extensibility in the circumferential direction is slightly greater for the hypertensive aortas as compared to normals. However, the stress-extension ratio relationship in the circumferential direction is similar in the two groups. 4. The stress-extension ratio relationship in the longitudinal direction indicates that the hypertensive aorta is stiffer than its normotensive counterpart.     

14. Are the impacts of events in the earth's history discernable in the current distributions of freshwater algae?

Freshwater microalgae, lacking a fossil record, have contributed little to the study of historical biogeography. Some of the innate difficulties are discussed, as well as some of the more hopeful possibilities, if distribution records, morphology and DNA sequence analysis are combined with knowledge of the earth's history. Examples of species within the same family showing quite different distributions are given, along with suggested explanations. These include possible examples of the role played by waterfowl in dissemination of freshwater algae.     

Detoxification of xenobiotics by plant cells: characterisation of vacuolar amphiphilic organic anion transporters

The transport activities of two primary ATP-dependent organic-anion transporters in the tonoplast of isolated barley (Hordeum vulgare L. cv. Klaxon) vacuoles have been characterised with N-ethylmaleimide glutathione (NEM-SG) and taurocholate as substrates. The transporters showed different sensitivities to organic anions and a variety of transport inhibitors and drugs. The vacuolar uptake of NEM-SG was inhibited by carbonylcyanide 4-trifluoromethoxyphenylhydrazone, 4,4-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS), S-(2,4-dinitrophenyl)glutathione, alkyl-S-glutathione derivatives and taurocholate but stimulated by probenecid. The uptake of taurocholate was inhibited by vinblastine, DIDS and probenecid. Both transporters were unaffected by verapamil. The kinetic properties of the transporters indicate a general preference for amphiphilic anions with some substrate overlap. These characteristics of the transporters are similar to those displayed by the multidrug resistance protein of mammalian drug-resistant cells. We suggest that these vacuolar transporters be described as plant multispecific organic anion transporters (pMOATs).Abbreviations Bm-S bimane S-glutathione - DIDS 4,4-diisothiocyanatostilbene-2,2-disulfonic acid - DNP-SG S-(2,4-dinitrophenyl)glutathione - FCCP carbonylcyanide 4-trifluoromethoxyphenylhydrazone - LTC₄ cysteinyl leukotriene - MDR multidrug transporter - MRP multidrug resistance protein - NEM-SG N-ethylmaleimide glutathione We thank Prof E. Martinoia for technical advice on the uptake experiments and Prof J. Palmer for helpful discussions and suggestions. M.B.-K. was partially sponsored by a grant from Stichting VSB Fonds, The Netherlands. IACR receives grant-aided support from the Biotechnology and Biological Science Research Council of the United Kingdom     

Growth of food-borne pathogenic bacteria in oil-in-water emulsions: I—Methods for investigating the form of growth

Methods are presented for investigating the site and form of growth of bacteria in model oil-in-water emulsions and in dairy cream. Following growth of the bacteria, the continuous aqueous phase is gelled using agarose and the oil phase removed using a mixture of chloroform and methanol. Using this method, the authors have found that Listeria monocytogenes, Salmonella typhimurium and Yersinia enterocolitica grow in the form of colonies in concentrated oil-in-water emulsions. Colonies of L. monocytogenes and Y. enterocolitica also form in artificially-inoculated fresh and tinned dairy cream. If information about the precise site of growth is not required, the authors have discovered that intact colonies can be liberated from the model emulsions by dissolving away the oil phase with chloroform: methanol.