Size fractionation of Trypanosoma brucei DNA: localization of the 177-bp repeat satellite DNA and a variant surface glycoprotein gene in a mini-chromosomal DNA fraction

We have size-fractionated intact DNA from Trypanosoma brucei into a major large DNA fraction (greater than 350S) and minor middle-sized (60-250S) and small (less than 60S) DNA fractions. Large DNA contains the rRNA genes, the basic copy genes for several variant surface glycoproteins (VSGs), including one which lies near a telomer, and the expression-linked copies of the two VSG genes. The middle-sized DNA contains at least one VSG gene, but the hybridization of this fraction with probes for the conserved repetitive sequences that mark the edges of the transposed segments of VSG genes, suggests that it may contain many VSG genes. The 177-bp repeat satellite DNA is also exclusively found in this fraction.     

Functional analysis of the Lactococcus lactis usp45 secretion signal in the secretion of a homologous proteinase and a heterologous α-amylase

The ups45 gene encodes the major extracellular protein from Lactococcus lactis. The deduced sequence of the 27 residue leader peptide revealed the tripartite characteristics of a signal peptide. This leader peptide directed the efficient secretion of the homologous proteinase (PrtP) in L. lactis, indicating that the putative signal peptide of PrtP can be replaced by the 27 residue Usp45 leader peptide. In addition, the 27 residue leader peptide could be used to secrete the Bacillus stearothermophilus α-amylase, encoded by the amyS gene. Fusion of the usp45 promoter region and various parts of the leader sequence to an amyS gene devoid of its signal sequence, showed that in Escherichia coli the first 19, 20, and 27 residues of the Usp45 leader are able to direct α-amylase secretion. In L. lactis the shorter signal peptides did not result in secretion of α-amylase, providing experimental evidence for the hypothesis that gram-positive bacteria require a longer signal peptide for secretion than gram-negative organisms.     

The pelagic system in the photic zone of the tropical Atlantic

Summary A multidisciplinary study of the carbon budget in the upper 300 meter of permanently stratified waters was started by two NECTAR-expeditions (North Equatorial Current Trans Atlantic Research) with HMS TYDEMAN in 1977 and 1978 (BAARS, ZIJLSTRA and TIJSSEN, 1979). In 1979 additional measurements were performed during the Gulf of Guinea-expedition with MS TYRO.Primary production in the nutrient depleted mixed layer of the North Equatorial Current estimated from the diurnal cycle in the O₂ concentration (TIJSSEN, 1979) and in POC (POSTMA and ROMMETS, 1979) revealed values 4–10 times higher than the data from the¹⁴C method in the literature: 800–2000 against ca. 200 mg C/m²/day. Moreover,¹⁴C incubations performed in bottle volumes of 4 liter and over 2 hour periods, instead of the recommended 12 hours for oligotrophic waters, gave 5–15 times higher values as incubations in the commonly used 300 ml (or smaller) bottles (GIESKES, KRAAY and BAARS, 1979). In the latter bottles a dramatic decrease of chlorophyll concentrations was observed, suggesting either heavy damage to fragile microflagellates by glasswall contacts and/or insufficient nutrient recycling by the lack of zooplankton in small samples. This then could account for the phenomena of low production and decreasing algal stock in long incubations with small bottles. These results suggest that today's picture of the primary production in the world's oceans (DE VOOIJS, 1979) needs probably a thorough reexamination. On the other hand, experiments in the North Sea and in the Gulf of Guinea did not indicate an effect of bottle size on¹⁴C incorporation and the¹⁴C method gave comparable estimates of primary production as the high precision oxygen method (photometric endpoint detection in the Winkler titration,cf. TIJSSEN, 1979). However, in these waters nutrient concentrations are always clearly above detection levels so that all previous data from somewhat richer areas may have been correct. In relation to the large oligotrophic parts of the oceans a question remains about the fate of the probably high primary production. Is it consumed by the algae themselves by night (POSTMA, 1980), are bacteria and microzooplankton more important consumers than formerly thought, or is the daily ration of zooplankters much higher as expected from extrapolation of filtration experiments in more eutrophic waters? We hope to clarify this topic during the coming NECTAR'82 expedition.Another main objective of the programme concerns the character of the deep chlorophyll maximum in permanently stratified waters. This layer is in the North Equatorial Current located at the depth of the nitratocline and at ca. 1% of the incident light at the surface (SPITZER and WERNAND, 1981). Primary production in the layer seems to be negligible while detailed vertical profiles of zooplankton, obtained with a Longhurst-Hardy Plankton Recorder, revealed no obvious concentrations in the layer. Chlorophyll analysis by thin-layer-chromatography (GIESKES, KRAAY and TIJSSEN, 1978) demonstrated that more than half of the chlorophyll a in the layer consists of an isomer which bleaches rapidly when transferred to higher light levels. The hypothesis was formulated that the chlorophyll maximum layer, at least in this region of the Atlantic, is an accumulation of chlorophyll breakdown products with a quite long turnover time at low light levels. In contrast to these findings, chlorophyll maxima near West-Africa and in the Gulf of Guinea were located at 5–10% light and contained more living algal cells (most of them as small as 1–3 m) than the nutrientpoor mixed layer above it. Primary production profiles had therefore a bimodal shape, with peaks at 30–50% light and at the chlorophyll maximum. Chlorophyll isomers, now analysed with High Pressure Liquid Chromatography, were also in these waters present at nearly all stations and comprised 30–70% of total chlorophyll comparable to the situation in the North Equatorial Current (GIESKES and KRAAY, 1981). It could be shown that these isomers are not involved in primary production, so that well established relationships between chlorophyll, light and primary production, found in oceanographic literature, have to be reconsidered.     

A Weight of Evidence Approach to the Aquatic Hazard Assessment of Bisphenoi A

Bisphenol A (BPA; 4,4-isopropylidene diphenol) is a chemical intermediate used primarily in the production of epoxy resins and polycarbonate products. BPA has been identified in surface waters and, hence, has been the subject of considerable research into its potential effects on aquatic organisms. Available literature on the aquatic toxicity of BPA was reviewed for quality against European Union TGD and Organization of Economic Cooperation and Development GLP principles. From this review, studies of suitable quality covering numerous ecologically relevant endpoints were identified to evaluate the survival, growth, and reproductive success of aquatic organisms exposed to BPA. Those studies yielded approximately 70 no observed effect concentrations (ranging from 16 to 3640 μg/L) and lowest observed effect concentrations (160 to 11,000 μg/L) that were considered in this weight of evidence assessment. Across all data, adverse effects on survival, growth, and reproduction occurred only at concentrations of 160 μg/L and above. Secondary biochemical (e.g., vitellogenin induction) and morphological (e.g., gonad histology) data provide insight into mechanisms of action, but do not correlate with apical endpoints related to survival, growth, and reproduction. Comparing the weight of the evidence of the aquatic toxicity data that showed chronic effects at 160 μg/L and higher with typical surface water concentrations in the range of 0.001 to 0.10 μg/ L, BPA is unlikely to cause adverse effects on aquatic populations or ecosystems.     

Der Einfluß der Elbe auf die Verunreinigung der Nordsee

Zusammenfassung 1. Der Zustrom von organischem Restmaterial aus den Flüssen in das Meer stellt eine Art von Verschmutzung dar mit der Folge einer Erhöhung des Trophiespiegels. Der Umfang der Verdünnung im Vorfluter und allgemein die geographischen und hydrographischen Situationen bestimmen das aus dem Angebot an Nährstoffen resultierende marine Equilibrium.2. Im Elbe-Aestuar haben wir alle Übergänge zwischen mixo-oligohalinen und -polyhalinen Zonen, deren Lage in den Jahreszeiten und zum Teil kurzfristig stark verschoben werden kann. Dadurch ergeben sich für das Benthos extrem astatische Verhältnisse.3. Durch den Tidewechsel wird das Plankton in den einzelnen Halinitätsbereichen länger zurückgehalten als in einem einseitig abfließenden Strom. Entsprechend dieser verlängerten Verweilzeit können sich typische Plankton-Biocoenosen in regionaler Sukzession ausbilden. Das Phytoplankton mit seiner schnellen Reproduktionsrate ist hierbei besonders begünstigt.4. Aus hydrographischen Gründen entsteht im oligo- bis mesohalinen Bereich eine Trübungszone, deren Material vornehmlich aus den Resten der limnischen Produktion (und Tonmineralien) besteht. Durch die natürlichen Gegebenheiten ist ein Aestuar-System sozusagen auf eine gesteigerte Dekompositionsleistung eingestellt, so daß eine anthropogene Zugabe von organischem Restmaterial ohne stärkere Beeinträchtigung des biocoenotischen Systems möglich ist.5. Im Vormündungsgebiet der Elbe bewirkt der Einstrom großer Mengen gelöster Nährstoffe aus dem Aestuar eine Erhöhung des trophischen Niveaus und eine Sekundärverschmutzung, deren Auffangzonen die tieferen Meeresflächen und die Wattenbereiche darstellen. Die Watten sind biocoenotisch auf die Aufnahme großer Mengen an Detritusmaterial eingestellt; es wird die Frage diskutiert, wieweit in solche Bereiche auch eine zusätzliche Einleitung von organischem Abwasser ohne wesentliche Beeinträchtigung des biologischen Equilibriums möglich ist.

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Influence of the elbe on the pollution of the north sea

The inflow of organic rest material from rivers into the sea represents a kind of pollution resulting in an increased trophic level. Dilution in the river as well as the general geographic and hydrographic situations determine the marine equilibrium resulting from the supply of nutrients. In the Elbe estuary exist all kinds of transitions between mixo-oligohaline and mixo-polyhaline zones; the location of which may shift over decades or, in part, over short periods of time. As a consequence, the benthos is exposed to extreme variations. Due to the alternation of tides, the plankton remains longer within the individual salinity zones than would be the case in a one directional flow system. Hence typical planktonic biocoenoses can develop in regional successions. In this regard the phytoplankton with its fast reproduction rates is especially favoured. Due to hydrographical reasons a turbidity zone is formed in the oligoto merohaline areas containing primarily remains of the limnic production. An estuary system is, by its nature, adjusted to accept increased decomposition loads; addition of organic anthropogenic remains is, therefore, tolerable without significant biocoenotic consequences. The inflow of dissolved nutrients from the Elbe results in an increase of the trophic niveau and a secondary pollution in the outer regions of the estuary. Mud flats and deeper sea areas are capable of accepting large amounts of detritus. The question, to what extent these areas may be capable of tolerating additional organic waste products without negative biological consequences, is discussed.

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Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Fine mapping and imprinting analysis for fatness trait QTLs in pigs

Quantitative trait loci (QTL) for fatness traits were reported recently in an experimental Meishan × Large White and Landrace F₂ cross. To further investigate the regions on pig Chr 2 (SSC2), SSC4, and SSC7, 25 additional markers from these regions were typed on 800 animals (619 F₂ animals, their F₁ parents, and F₀ grandfathers). Compared with the published maps, a modified order of markers was observed for SSC4 and SSC7. QTL analyses were performed both within the half-sib families as well as across families (line cross). Furthermore, a QTL model accounting for imprinting effects was tested. Information content could be increased considerably on all three chromosomes. Evidence for the backfat thickness QTL on SSC7 was increased, and the location could be reduced to a 33-cM confidence interval. The QTL for intramuscular fat on SSC4 could not be detected in this half-sib analysis, whereas under the line cross model a suggestive QTL on a different position on SSC4 was detected. For SSC2, in the half-sib analysis, a suggestive QTL for backfat thickness was detected with the best position at 26 cM. Imprinting analysis, however, revealed a genome-wise, significant, paternally expressed QTL on SSC2 with the best position at 63 cM. Our results suggest that this QTL is different from the previously reported paternally expressed QTL for muscle mass and fat deposition on the distal tip of SSC2p. Received: 15 October 1999 / Accepted: 21 February 2000     

ICOS expression by activated human Th cells is enhanced by IL-12 and IL-23: increased ICOS expression enhances the effector function of both Th1 and Th2 cells

Previous mouse studies have shown that IL-4 increases the expression of ICOS on activated Th cells, resulting in enhanced ICOS expression on Th2 cells. In this study, we show that ICOS expression on human Th cells is not increased by IL-4, but by IL-12 and by IL-23 instead. Consequently, ICOS expression during IL-12-driven Th1 cell polarization was transiently increased compared with the levels on Th0 cells and IL-4-driven Th2 cells. Addition of IL-12 and/or IL-23 during restimulation increased ICOS expression to the same extent on pre-established Th1, Th2, and Th0 cells, indicating that ICOS levels are not stably imposed by prior polarization. In contrast to the findings in the mouse, IL-4 significantly suppressed the ICOS-enhancing effects of IL-12 and IL-23. The functional consequence of variable ICOS levels was shown in coculture experiments with cells expressing the ICOS-ligand B7-related protein 1 (either transfected Chinese hamster ovary cells or autologous dendritic cells). Ligation of ICOS on 2-day-preactivated effector cells increased their cytokine production to an extent proportional to their ICOS expression levels. As the ICOS-enhancing potentials of IL-12 and IL-23 were maintained for several days after stimulation, both on Th1 and Th2 cells, we propose the concept that local regulation of ICOS expression on activated Th cells by IL-12 and/or IL-23 may provide a powerful means to amplify effector T cell responses in peripheral tissues, independently of the polarized state of the Th cells.