GENETIC AND MORPHOLOGICAL VARIATION OF POPULATIONS BELONGING TO THE BULINUS TRUNCATUS/TROPICUS COMPLEX (GASTROPODA; PLANORBIDAE) IN SOUTH WESTERN ZIMBABWE

Aquatic snails from south western Zimbabwe belonging to theBulinus trunscatus/tropicus complex vary widely in shell formsuggesting that more than one taxon could be present. This possibilitywas investigated by making observations on snail samples from13 populations from the Plumtree area, in respect of allozymevariation (5 polymorphic loci), shell morphology (9 variables),copulatory organ and chromosome number. Comparative data wereobtained from snails from north western Zimbabwe identifieddefinitely as B. tropicus. Analysis of the genetic structurerevealed a high degree of polymorphism (P) ranging from 0.29–0.80among populations from Plumtree and expected heterozygosity(H_e) from 0.02–0.22. No enzymatic diagnostic loci werefound which could differentiate the different morphs or populationsand discriminant function analysis on the morphological datashowed an overlap of morphs among populations. Snails analyzedfor chromosome number were all diploid (2n = 36). Snails exposedto Schistosoma haematobium mira-cidia were all refractory. Thisinformation supports the view of a single species, B. tropicus,which is differentiated due to migration barriers and whereenvironmental variables might be implicated in the morphometricdivergence. (Received 31 July 1995; accepted 15 January 1998)     

Above- and Below-ground Production of Young Scots Pine (Pinus sylvestris L.) Trees after Three Years of Growth in the Field under Elevated CO2

Scots pine (Pinus sylvestris L.) seedlings were grown for 3years in the ground in open top chambers and exposed to twoconcentrations of atmospheric CO₂(ambient or ambient + 400 µmol mol^-1) without addition of nutrients and water. Biomassproduction (above-ground and below-ground) and allocation, aswell as canopy structure and tissue nitrogen concentrationsand contents, were examined by destructive harvest after 3 years.Elevated CO₂increased total biomass production by 55%, reducedneedle area and needle mass as indicated, respectively, by lowerleaf area ratio and leaf mass ratio. A relatively smaller totalneedle area was produced in relation to fine roots under elevatedCO₂. The proportion of dry matter in roots was increased byelevated CO₂, as indicated by increased root-to-shoot ratioand root mass ratio. Within the root system, there was a significantshift in the allocation towards fine roots. Root litter constituteda much higher fraction of fine roots in trees grown in the elevatedCO₂than in those grown in ambient CO₂. Growth at elevated CO₂causeda significant decline in nitrogen concentration only in theneedles, while nitrogen content significantly increased in branchesand fine roots (with diameter less than 1 mm). There were nochanges in crown structure (branch number and needle area distribution).Based upon measurements of growth made throughout the 3 years,the greatest increase in biomass under elevated CO₂took placemainly at the beginning of the experiment, when trees grownin elevated CO₂had higher relative growth rates than those grownunder ambient CO₂; these differences disappeared with time.Symptoms of acclimation of trees to growth in the elevated CO₂treatmentwere observed and are discussed. Copyright 2000 Annals of BotanyCompany Elevated CO₂, Pinus sylvestris, biomass production, allocation, fine roots, root litter, crown structure, nitrogen, C/N ratio     

The role of Asn14 in the stability and conformation of the reactive-site loop of winged bean chymotrypsin inhibitor: crystal structures of two point mutants Asn14-->Lys and Asn14-->Asp.

A double-headed chymotrypsin inhibitor, WCI, from winged bean seeds was cloned for structural and biochemical studies. The inhibitor was subjected to two point mutations at a conserved position, Asn14. This residue, known to have a pivotal role in stabilizing the first reactive-site loop (Gln63-Phe68) of the inhibitor, is highly conserved in the sequences of the other members of Kunitz (STI) family as well as in the sequences of Kazal family of serine protease inhibitors. The mutants, N14K and N14D, were subjected to biochemical assay and their characteristics were compared with those of the recombinant inhibitor (rWCI). Crystallographic studies of the recombinant and the mutant proteins are discussed. These studies were primarily aimed at understanding the importance of the protein scaffolding towards the conformational rigidity of the reactive-site loop. Our analysis reveals that, as the Lys14 side chain takes an unusual fold in N14K and the Asp14 side chain in N14D interacts with the loop residues by water-mediated hydrogen bonds, the canonical conformation of the loop has remained effectively intact in both the mutant structures. However, minor alterations such as a 2-fold increase in the inhibitory affinity towards the cognate enzyme were observed.     

Effect of acute exposure to 3660 m altitude on orthostatic responses and tolerance.

Orthostatic reflexes were examined at 375 m and after 60 min of exposure in a hypobaric chamber at 3660 m using a 20-min 70 degrees head-up tilt (HUT) test. Mean arterial blood pressure, R wave-R wave interval (RRI), and mean cerebral blood flow velocity (MFV) were examined with coarse-graining spectral analysis. Of 14 subjects, 7 at 375 m and 12 at 3660 m were presyncopal. Immediately on arrival to high altitude, breathing frequency and MFV increased, and endtidal PCO2, RRI, RRI complexity, and the parasympathetic nervous system indicator decreased. MFV was similar in HUT at both altitudes. The sympathetic nervous system indicator increased with tilt at 3660 m, whereas parasympathetic nervous system indicator decreased with tilt at both altitudes. Multiple regression analysis of supine variables from either 375 or 3660 m and the time to presyncope at 3660 m indicated that, after 1 h of exposure, increased presyncope at altitude was the result of 1). ineffective peripheral vasoconstriction, despite increased cardiac sympathetic nervous system activity with HUT, and 2). insufficient cerebral perfusion owing to cerebral vasoconstriction as the result of hypoxic hyperventilation-induced hypocapnia.     

Lysosomal accumulation and recycling of lipopolysaccharide to the cell surface of murine macrophages, an in vitro and in vivo study.

In this study, we detailed in a time-dependent manner the trafficking, the recycling, and the structural fate of Brucella abortus LPS in murine peritoneal macrophages by immunofluorescence, ELISA, and biochemical analyses. The intracellular pathway of B. abortus LPS, a nonclassical endotoxin, was investigated both in vivo after LPS injection in the peritoneal cavity of mice and in vitro after LPS incubation with macrophages. We also followed LPS trafficking after infection of macrophages with B. abortus strain 19. After binding to the cell surface and internalization, Brucella LPS is routed from early endosomes to lysosomes with unusual slow kinetics. It accumulates there for at least 24 h. Later, LPS leaves lysosomes and reaches the macrophage cell surface. This recycling pathway is also observed for LPS released by Brucella S19 following in vitro infection. Indeed, by 72 h postinfection, bacteria are degraded by macrophages and LPS is located inside lysosomes dispersed at the cell periphery. From 72 h onward, LPS is gradually detected at the plasma membrane. In each case, the LPS present at the cell surface is found in large clusters with the O-chain facing the extracellular medium. Both the antigenicity and heterogenicity of the O-chain moiety are preserved during the intracellular trafficking. We demonstrate that LPS is not cleared by macrophages either in vitro or in vivo after 3 mo, exposing its immunogenic moiety toward the extracellular medium.