Micronekton and macrozooplankton in the open waters near Antarctic ice edge zones (AMERIEZ 1983 and 1986)

Summary Micronekton and macrozooplankton assemblages (0–1000 m) were sampled from the open ocean in the vicinity of marginal ice zones in the southern Scotia and western Weddell Seas using midwater trawls. Small regional differences in species composition were found in the differing hydrographic settings with the Scotia Sea being slightly more diverse. Most species exhibited broad vertical ranges with no distinct pattern of vertical movement. Exceptions were mesopelagic fish and Salpa thompsoni which undertook diel vertical migrations. Biomass was high (2.4–3.1 g DW/m²), comparable to Pacific subarctic waters. Euphausia superba and Salpa tompsoni were the numerical and biomass dominants, representing over 50% of the total numbers and standing stocks. In terms of biomass, euphausiids were the most important group at shallow depths (0–200 m) but were surpassed by salps in the Scotia Sea and mesopelagic fish in the Weddell Sea when all depths down to 1000 m were considered. Pelagic fish biomass (3.3–4.4 g WW/m²) greatly exceeded published estimates for birds (0.025–0.070 g WW/m²), seals (0.068–0.089 g WW/m²) and whales (0.167 to 0.399 g WW/m²), making mesopelagic fish the most prevalent krill predators in the Antarctic oceanic system.     

Bacterial expression of an active class Ib chitinase from Castanea sativa cotyledons

Ch3, an endochitinase of 32 kDa present in Castanea sativa cotyledons, showed in vitro antifungal properties when assayed against Trichoderma viride. The characterization of a cDNA clone corresponding to this protein indicated that Ch3 is a class Ib endochitinase that is synthesized as a preprotein with a signal sequence preceding the mature polypeptide. Bacterial expression of mature Ch3 fused to the leader peptide of the periplasmic protein ompT resulted in active Ch3 enzyme. A plate assay was adapted for semi-quantitative determination of chitinase activity secreted from cultured bacteria, which should facilitate the identification of mutants with altered capacity to hydrolyse chitin.     

Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes.

PR1 is a pathogenesis-related protein encoded in the parsley genome by a family of three genes (PR1-1, PR1-2 and PR1-3). Loss- and gain-of-function experiments in a transient expression system demonstrated the presence of two fungal elicitor responsive elements in each of the PR1-1 and PR1-2 promoters. These elements, W1, W2 and W3, contain the sequence (T)TGAC(C) and mutations that disrupt this sequence abolish function. Gel shift experiments demonstrated that W1, W2 and W3 are bound specifically by similar nuclear proteins. Three cDNA clones encoding sequence-specific DNA-binding proteins were isolated by South-Western screening and these proteins, designated WRKY1, 2 and 3, also bind specifically to W1, W2 and W3. WRKY1, 2 and 3 are members of the family of sequence-specific DNA-binding proteins, which we call the WRKY family. Treatment of parsley cells with the specific oligopeptide elicitor Pep25 induced a transient and extremely rapid increase in mRNA levels of WRKY1 and 3. WRKY2 mRNA levels in contrast showed a concomitant transient decrease. These rapid changes in WRKY mRNA levels in response to a defined signal molecule suggest that WRKY1, 2 and 3 play a key role in a signal transduction pathway that leads from elicitor perception to PR1 gene activation.     

Genetic mapping of new morphological, isozyme and RAPD markers in Vicia faba L. using trisomics

Thirteen F₂ families of faba bean (Vicia faba L.), descended from plants trisomic for chromosomes 3, 4, 5 and 6, have been analyzed for morphological, isozyme and RAPD markers. This allowed the establishment of linkage relationships among these markers as well as the assignment of some markers and/or linkage groups to their respective chromosomes. The linkage analysis of partially overlapping sets of informative genetic markers for the data pooled from 13 F₂ families has revealed 48 linkage groups, six of which have been precisely assigned to specific chromosomes. A statistical procedure to analyze the data of joint segregation analysis in families derived from trisomic plants has been developed.     

An extended x-ray absorption fine structure study of the high-affinity cation-binding site in the purple membrane.

The structure of the high-affinity cation-binding site of bacteriorhodopsin was studied using extended x-ray absorption fine structure techniques. The results obtained for Mn2+ in aqueous solution and for the complex BR-Mn2+ (1:1 molar ratio) show great similarities, suggesting that Mn2+, when bound to this site, is coordinated with six atoms of oxygen, forming an octahedral disposition. The interatomic distance between the atoms of oxygen and the Mn2+ was found to be 2.17 A for the complex BR-Mn2+, similar to Mn2+ in solution (2.15 A). In addition, the absence of any other peak at greater distances in the Fourier-transformed spectrum indicates that neither phosphorus nor sulphur atoms are present in the second coordination shell. This suggests that this binding site is located in the protein, discarding the proximity of lipid polar headgroups.     

Large scale production of recombinant mouse and rat growth hormone by fed-batch GS-NSO cell cultures

Investigations of biological effects of prolonged elevation of growth hormone in animals such as mice and rats require large amounts of mouse and rat growth hormone (GH) materials. As an alternative to scarce and expensive pituitary derived materials, both mouse and rat GH were expressed in NSO murine myeloma cells transfected with a vector containing the glutamine synthetase (GS) gene and two copies of mouse or rat GH cDNA. For optimal expression, the mouse GH vector also contained sequences for targeting integration by homologous recombination. Fed-batch culture processes for such clones were developed using a serum-free, glutamine-free medium and scaled up to 250 L production scale reactors. Concentrated solutions of proteins, amino acids and glucose were fed periodically to extend cell growth and culture lifetime, which led to an increase in the maximum viable cell concentration to 3.5×10⁹ cells/L and an up to 10 fold increase in final mouse and rat rGH titers in comparison with batch cultures. For successful scale up, similar culture environmental conditions were maintained at different scales, and specific issues in large scale reactors such as balancing oxygen supply and carbon dioxide removal, were addressed. Very similar cell growth and protein productivity were obtained in the fed-batch cultures at different scales and in different production runs. The final mouse and rat rGH titers were approximately 580 and 240 mg/L, respectively. During fed-batch cultures, the cell growth stage transition was accompanied by a change in cellular metabolism. The specific glucose consumption rate decreased significantly after the transition from the growth to stationary stage, while lactate was produced in the exponential growth stage and became consumed in the stationary stage. This was roughly coincident with the beginning of ammonia and glutamate accumulation at the entry of cells into the stationary stage as the result of a reduced glutamine consumption and periodic nutrient additions.     

Adenovirus type 9 E4 open reading frame 1 encodes a transforming protein required for the production of mammary tumors in rats.

The E4 region of human adenovirus type 9 (Ad9) transforms established rat embryo fibroblasts and encodes an essential determinant for the production of estrogen-dependent mammary tumors in rats. Testing of the seven Ad9 E4 open reading frames (ORFs) individually for transformation of the established rat embryo fibroblast cell line CREF indicated that only Ad9 E4 ORF1 possessed a significant ability to generate transformed foci on these cells. In contrast, the E4 ORF1 sequences from human Ad5 and Ad12 lacked the transforming potential exhibited by Ad9 E4 ORF1. Cell lines derived from Ad9 E4 ORF1-transformed foci expressed the 14-kDa Ad9 E4 ORF1 protein and formed colonies in soft agar. In addition, the Ad9 E4 ORF1 protein was required for initiation of mammary oncogenesis in vivo, as E4 ORF1 mutant viruses failed while E4 ORF2 and ORF3 mutant viruses succeeded in eliciting mammary tumors in animals. A role for Ad9 E4 ORF1 in tumor maintenance was suggested by the fact that 100% of virus-induced mammary tumors expressed the E4 ORF1 protein. Taken together, the facts that the Ad9 E4 ORF1 protein exhibits transforming potential in culture and is required by Ad9 to produce mammary tumors in animals suggest that Ad9 E4 ORF1 is a new viral oncoprotein.     

Bats of the Gulf of Guinea islands: faunal composition and origins

The present study compares the bat faunas of the islands of the Gulf of Guinea. Species composition. endemism and hypothetical origins are discussed. All families present in the mainland region are found in Bioko, a typical landbridge island. Foliage gleaning guild species (Nycteridae) show limited colonization abilities. This is also true of the family Rhinolophidae, but not for the closely related family Hipposideridae. The majority of the oceanic island species are African bats which show a widespread distribution and, therefore, have a high ecological plasticity. The continental relatives of the two endemic species Myonycteris brachycephala and Chaerephon tomensis are restricted to relatively small forested areas. Bioko's bat fauna is the result of the recent isolation from a formerly land-connected community. The oceanic bat faunas originated from the establishment of incomers from other areas. Nevertheless, extinction appears in both vicariant and dispersal processes, as an important factor in modelling the current bat communities of the Gulf of Guinea islands.     

Chemical composition of antarctic zooplankton during austral fall and winter

Water level, ash content, proximate (protein, lipid, carbohydrate and chitin) and elemental (carbon and nitrogen) composition were analyzed in twentythree species of Antarctic Zooplankton collected during the austral fall (1986) and winter (1988) from the Scotia/Weddell Sea region. Extremes in water level, ash content and organic components were typified by copepods and gelatinous forms. Ostracods and polychaetes were generally similar in composition to copepods, being only slightly higher in water level and ash content. Chaetognaths exhibited a composition intermediate in character with some components similar in value to that shown by crustaceans (i.e. protein) while other components were more in the range of values seen in gelatinous forms (i.e. water level and ash content). Protein was the major proximate component and measured values (as % Afdw) were fairly uniform among non-gelatinous species (x=33.9±6.9). Lipid levels were variable, with high values (>30% AFDW) only found for the copepods Calanoides acutus, Calanus propinquus and Euchaeta antarctica. Carbohydrate values were low in all species examined. Chitin was measured in crustacean species only. With the exception of C. acutus (x=2.5% AFDW chitin), values were similar among species with mean values being slightly higher in fall (x=11.8±2.5) than in winter (x=6.7±1.8). Among non-gelatinous species, the ratio of carbon to nitrogen was positively correlated with the lipid to protein ratio, underscoring the compositional association between elemental and proximate components in these groups. In gelatinous species, the relationship between carbon:nitrogen and lipid:protein was inconsistent and less pronounced. Caloric content was estimated from recovered organic matter for nongelatinous species. As a function of wet weight and dry weight, values reflected differences in water level and ash content among individual species. As a function of ashfree dry weight, values were similar among all species (x=3.6±0.9 kcal/g).Seasonal comparisons were possible for 12 of the 23 species. Among crustaceans, changes in water level and organic components were variable reflecting dissimilar trophic, reproductive or ecological habits among different species. Essentially no change in composition between fall and winter was observed for diapause species (e.g. Calanoides acutus and Rhincalanus gigas) as well as for omnivorous/ carnivorous species (e.g. Gaetanus tenuispinus). Conversely, large compositional changes were evident for Calanus propinquus, a small-particle grazer that relies heavily on lipid reserves. Chaetognaths and some gelatinous species exhibited a considerable decrease in ash content from fall to winter which, for most cases, was mirrored by some degree of increase in lipid level. At present, however, scant data are available to help explain the observed patterns of compositional change within non-crustacean species.     

Adenylate Uptake by Proplastids from Cultured Cells of Tobacco {Nicotiana tabacum L. cv. BY2) Indicates That an Adenylate Translocator is Present in All Types of Plastid

The presence of an adenylate translocator in the envelope membranesof proplastids isolated from the cultured cells of tobacco (Nicotianatabacum L. cv. BY2) was examined by means of transport experimentsusing the silicone oil filtering centrifugation technique. Itwas observed that proplastids can import [³H]ATP, [³H]ADP, [³H]AMPand less specifically ADP-[¹⁴C]Glc which can eventually be usedfor starch biosynthesis. The effects of specific inhibitorsof the mitochondrial adenylate translocator, i.e. atractyloside,bongkrekic acid and carboxyatractyloside were tested. Similarto the case of amyloplasts isolated from the cultured cellsof sycamore and chloroplasts isolated from spinach leaves, onlyATP and ADP-Glc uptake were shown to be partially inhibitedby carboxyatractyloside. On the other hand, neither atractylosidenor bongkrekic acid exerted a significant inhibitory effecton adenylate uptake. (Received August 8, 1992; Accepted November 26, 1992)