Chemically-Synthesized Gene Encoding Modified Human Superoxpde Dismutase: Its Construction,Expression and Properties of the Product

The gene encoding modified human superoxide dismutax (h-SOD) with 153 amino acid residues was constructed by chemical synthesis using the phosphoramidite method. The gene was designed so as to use bacterial codons for expression in prokaryotes and to introduce several unique restriction sites for further mutagenesis by the cassette exchange method. The distance between Shine-Dalgarno sequence and initiation codon was adjusted to maximum expression by using synthesized oligonucleotide. In addition, Cys 6 of h-SOD was changed to Ala to improve instability of native h-SOD.

Synthesized structural gene of h-SOD was expressed in E. coli after induction of isopropyl β-D-thiogalactoside by inserting the gene into the expression vector pKK223–3 having tac promoter. The gene that has 10 base pairs between Shine-Dalgarno sequence and initiation codøn showed the most efficient expression. The gene produced three active SOD isomers as revealed by chromatofocusing.

The main isomer was purified to homogeneity and characterized. The h-SOD-Ala⁶ showed similar properties to those of native h-SOD with respect to molecular weight, subunit structure, absorption spectrum. but the modified SOD was more resistant to heat denaturation than was native h-SOD; half-denaturing temperature was shifted by 10°C. Thus. the exchange of Cys 6 to Ala of h-SOD increased a stability of the enzyme.     

Flight behaviour of Encarsia formosa in response to plant and host stimuli

The flight behaviour of Encarsia formosa Gahan (Hymenoptera: Aphelinidae), a parasitoid of Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae) has been studied in response to the following natural components: bean plant (Phaseolus vulgaris L.) infested with T. vaporariorum (PHC, plant host complex), host damaged bean plant from which whiteflies were removed (HDP, host damaged plant), T. vaporariorum third instar larvae (H, host), uninfested bean plant (P, plant) and T. vaporariorum empty pupal cases (EPC, empty pupal cases). A combination of chemical and visual stimuli is necessary in host location from a distance by E. formosa and seems to deeply affect the dispersal of this parasitoid. The presence of host (PHC, H) or of its parts (EPC) led to a substantial increase of oriented flights suggesting the occurrence of host derived chemicals. At a closer range, a combination of visual and chemical stimuli deriving from all the components of the plant host complex elicited the transition flying-landing and the following landing on the source.     

Geographic variation of flavonoids in Phlox carolina

Non-anthocyanin flavonoid expression was determined among 64 continuous range and nine disjunct range populations of Phlox carolina. The species contains four flavone nuclei and a total of 25 derivatives, most of which are $\text{C-}\text{glycosides}$. Nearly all populations express unique flavone patterns. Using a qualitative similarity index of minimum biosynthetic step distance (MBSD), the average population contains only ca. 50% of the overall substitutional diversity. The separate ranges are distinctive in the % occurrence of most compounds but only weakly divergent in comparative mean MBSD. Proximity and MBSD are significantly correlated overall but local variability is extensive and no discrete racism is apparent. Within the continuous range 18 compounds are distributed in independent but overlapping gradients and 13 of these extend consistently through the disjunct range. The patch-work continuum of flavone variation is the apparent consequence of dissection and contraction of a historically more widespread distribution. Other general aspects of the variation patterns are discussed.     

Chemical composition of fresh and composted urban waste

Chemical, spectroscopic (IR and NMR), and molecular characteristics of humus extracted from urban waste before and after compositing are reported. The main differences are in the contents of acidic groups and straight aliphatic chains which diminish in the composted material. In comparing humus characteristics of composted urban waste and soils, the only real difference was found in the elution curves on Sephadex G-100, where the ratio of 100,000 and 1,300 peaks was higher in the compost than in the soil curve. The finding suggests that this ratio is a parameter that could enable us, even if in a semiquantitative manner, to follow the humification process of the compost when its addition to the soil is not less than the autochthonous organic matter content.Research work supported by CNR, Italy, Special grant I.P.R.A.-Sub-project 1. Paper N. 1531.     

Effect of streptolysin S on liposomes Influence of membrane lipid composition on toxin action

The effect of the bacterial cytolytic toxin, streptolysin S, on liposomes composed of various phospholipids was investigated. Large unilamellar vesicles containing [¹⁴C]sucrose were prepared by reverse-phase evaporation, and membrane damage produced by the toxin was measured by following the release of labeled marker. The net charge of the liposomes had little or no effect on their susceptibility to steptolysin S and the toxin was about equally effective on liposomes composed of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine and phosphatidylglycerol. Experiments with liposomes composed of synthetic phospholipids showed that the ability of the toxin to produce membrane damage depended on the degree of unsaturation of the fatty acyl chains. The order of sensitivity was C18 : 2 phosphatidylcholine > C18 : 1 phosphatidylcholine > C18 : 0 phosphatidylcholine = C16 : 0 phosphatidylcholine. Liposomes containing the latter two phospholipids were virtually unaffected by streptolysin S, and experiments with C18 : 0 phosphatidylcholine suggested that toxin activity does not bind to liposomes composed of phospholipids with saturated fatty acyl chains. The inclusion of 40 mol% cholesterol in C16 : 0 phosphatidylcholine and C18 : 0 phosphatidylcholine liposomes made these vesicles sensitive to streptolysin S. Egg phosphatidylcholine liposomes, which were unaffected at 0°C and 4°C became susceptible to the toxin at these temperatures when cholesterol was included. Liposomes composed of C14 : 0 phosphatidylcholine were unaffected by streptolysin S at temperatures below the chain-melting transition temperature (23°C) of this phospholipid, but became increasingly susceptible above this temperature. The results suggest that the fluidity of the phospholipid hydrocarbon chains in the membrane is important in streptolysin S action.     

Epicuticular wax of Cirsium arvense

Epicuticular wax of Cirsium arvense contains hydrocarbons (12%), esters (35%), free acids (3%), free alcohols (10%), triterpene acetates (8%) and 1,3-ditetradecanoyl-2-hexanoylglycerol (8%) as major components. Minor components are triterpene alcohols (3%) and nonacosan-10-ol (2%). The esters contain triterpene alcohol esters (19%) as well as esters of alkanols.     

Ribosomal Tag Pyrosequencing of DNA and RNA Reveals “Rare” Taxa with High Protein Synthesis Potential in the Sediment of a Hypersaline Lake in Western Australia

Little is known about the potential activity of microbial communities in hypersaline sediment ecosystems. Ribosomal tag libraries of DNA and RNA extracted from the sediment of Lake Strawbridge (Western Australia) revealed bacterial and archaeal operational taxonomic units (OTUs) with high RNA/DNA ratios providing evidence for the presence of ‘rare’ but potentially “active” taxa. Among the ‘rare’ bacterial taxa Halomonas, Salinivibrio and Idiomarina showed the highest protein synthesis potential. Rare but ‘active’ archaeal OTUs were related to the KTK 4A cluster and the Marine-Benthic-Groups B and D. We present the first molecular analysis of the microbial diversity and protein synthesis potential of rare microbial taxa in a hypersaline sediment ecosystem.     

Physical anthropological approaches to aging

The field of gerontology is remarkably diverse; yet there has been relatively little investigation of physical anthropological issues in aging research. This review explores gerontologic topics of actual and/or potential interest to physical anthropologists. The evolution of aging presents a theoretical dilemma in that postreproductively expressed traits may be outside the influence of natural selection. The physiological changes of aging comprise a diversified mosaic of physical deterioration as would be expected from an evolutionary model. Studies of prehistoric aging are limited to estimating lifespan, which may not be indicative of rate of aging. Considerable attention has been devoted to body composition and aging, and notable findings include a loss of lean tissue with age and relatively constant (though redistributing) fat mass. Osteoporosis is a major problem of aging in females, as is tooth loss in both sexes. The study of variation in rates of aging is only beginning, and a number of approaches to measuring biological age in adulthood are presented. Determining the associations of lifestyle, economic, and nutritional status with biological age may reveal sources of variation in rates of aging and may be of practical importance.     

Mechanisms of host discrimination and intraspecific competition in the aphid parasitoid Ephedrus cerasicola

The aphid parasitoid Ephedrus cerasicola Starý (Hymenoptera: Aphidiidae) discriminates between parasitized and unparasitized aphid hosts (Myzus persicae (Sulzer), Homoptera: Aphididae) in two different ways. By antennal contact with the parasitized host, it can detect an external marking pheromone which is effective only for a few hours. The behaviour of the parasitoid also indicated that it responded to an internal marker, probably by means of chemoreceptors on the ovipositor. The parasitoid behaviour was studied in encounters with parasitized hosts 0–10 min, 2–3 h, 6–7 h and 23–24 h after the first parasitization. Aphids were superparasitized within 1 h or after 24 h, and aphids in each experiment were later dissected at two different times. It is suggested that the supernumerary offspring were eliminated by chemical means, whether the time between the two ovipositions was 0–1 h or 24 h.

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Résumé Le parasitoïde de puceron, E. cerasicola Starý (Hym.: Aphidiidae) sélectionne les pucerons, Myzus persicae (Sulzer), parasités ou non, de deux façons différentes. Il peut percevoir une phéromone externe de marquage perceptible seulement pendant quelques heures par contact antennaire. Son comportement montre qu'il réagit aussi à un marquage interne, probablement par des chémorécepteurs de sa tarière. Le comportement du parasitoïde a été étudié en le mettant en contact avec des hôtes parasités depuis 0–10 min, 2–3 h, 6–7 h et 23–24 h. Les pucerons ont été surparasités pendant la première heure ou après 24 heures. Les pucerons de toutes les expériences ont été disséqués à deux moments différents. Ceci a conduit à estimer que les descendants en surnombre ont été éliminés chimiquement quand la période entre deux pontes successives était inférieure à 1 h ou supérieure à 24 h.

     

Differential Interactions Between an Aphid Endohyperparasitoid and Three Honeydew-Collecting Ant Species: A Field Study of Alloxysta brevis (Thomson) (Hymenoptera: Alloxystidae)

The behavioral interactions between the chemically protected aphid hyperparasitoid Alloxysta brevis and three trophobiotic ant species were examined in the field on thistles. The patterns were not essentially influenced by hyperparasitoid sex. Ants differed in their guarding behavior of aphids and their response toward A. brevis. Lasius niger attacked the hyperparasitoid more readily than Myrmica laevinodis, while aggression by Formica rufa was intermediate. Apart from applying chemical defense, A. brevis responded to ants by flying away (mainly at L. niger) or with avoidance behavior (mainly at M. laevinodis). Additionally, females might switch their tactics from flight to defense with increasing age. Females left the plant more often after encounters with L. niger or F. rufa than with M. laevinodis. Disturbance by any ant species affected the foraging activities, reducing oviposition success to nearly zero.