Anthropogenic impacts on the contextual morphological diversification and adaptation of Nile tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>, L. 1758) in East Africa

Nile tilapia occurs naturally in East Africa where it’s an economically important species. Many of the natural populations of Nile tilapia have been affected by anthropogenic activities including translocations, associated with programmes aimed at enhancing capture fisheries and aquaculture productivity. Using geometric morphometric analyses, we tested the hypothesis that such anthropogenic activities have augmented admixture among natural populations of Nile tilapia and influenced the geographical distribution of morphological variation within the species. Our expectation was that Nile tilapia shape divergent might be consistent with reportedly anthropogenic activities in nonnative environments. To test this hypothesis, we analyzed the shapes of 490 individuals from thirteen populations; three farms, six natives and four nonnative natural populations. Our analysis revealed that the most pronounced shape diversification was observed in seven populations; three nonnatives (Victoria, Kyoga and Sindi farm) and four natives (Albert, River Nile, George and Turkana). The features responsible for the observed morphotypes were mainly related to the orientation of the anterior region of the fish and may be due to diversifying selection in response to new environmental pressures (for nonnative populations), admixture or drift. Shape change in the nonnative high-altitude populations was unexpectedly conserved, suggesting recent introductions which may have not resulted in admixture or there was strong selection against change in the traits measured. On the other hand, the recorded morphotypic clusters explained the possible genetic link to their putative ancestral home. Our results were partially consistent with our prediction that the nonnative populations exhibited divergent morphotypes. We recommend further investigations with molecular genetics for follow-up of these findings.     

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

Antibodies against red blood cells (RBCs) can lead to complement activation resulting in an accelerated clearance via complement receptors in the liver (extravascular hemolysis) or leading to intravascular lysis of RBCs. Alloantibodies (e.g. ABO) or autoantibodies to RBC antigens (as seen in autoimmune hemolytic anemia, AIHA) leading to complement activation are potentially harmful and can be - especially when leading to intravascular lysis - fatal¹. Currently, complement activation due to (auto)-antibodies on RBCs is assessed in vitro by using the Coombs test reflecting complement deposition on RBC or by a nonquantitative hemolytic assay reflecting RBC lysis^1-4. However, to assess the efficacy of complement inhibitors, it is mandatory to have quantitative techniques. Here we describe two such techniques. First, an assay to detect C3 and C4 deposition on red blood cells that is induced by antibodies in patient serum is presented. For this, FACS analysis is used with fluorescently labeled anti-C3 or anti-C4 antibodies. Next, a quantitative hemolytic assay is described. In this assay, complement-mediated hemolysis induced by patient serum is measured making use of spectrophotometric detection of the released hemoglobin. Both of these assays are very reproducible and quantitative, facilitating studies of antibody-induced complement activation.     

Use of a murine secreted alkaline phosphatase as a non-immunogenic reporter gene in mice

BACKGROUND: The development of any vector system as a gene delivery system requires its optimization in vitro and in vivo. Preliminary studies frequently involve the use of a reporter gene, which allows for the rapid and simple assay of vector function through monitoring expression levels of the reporter gene. However, evaluation of vector efficacy can be compromised by immune responses directed against immunogenic reporter proteins. METHODS: We have cloned a murine secreted alkaline phosphatase (mSEAP), and explored its use as a reporter gene in the context of an early region 1 (E1)-deleted adenovirus (Ad) vector. Studies involved characterization of gene expression in vitro and in vivo, and immunological responses after gene delivery to mice. RESULTS: In tissue culture, we show that mSEAP is easily measured quantitatively using a sensitive, commercially available chemiluminescent assay, or visualized directly using histological staining. The level of transgene expression from AdmSEAP was similar to that observed for an Ad vector encoding the human placental secreted alkaline phosphatase (hSEAP). After intravenous administration in mice, AdmSEAP continued to express at high levels for the duration of the experiment (1 month), whereas expression from AdhSEAP declined to background levels over the course of the experiment. Although cytotoxic T-lymphocytes were not detected against either the murine or human SEAP proteins in mice, antibodies were readily detected against the human protein. No antibodies were detected to mSEAP. CONCLUSIONS: Taken together, these data illustrate that mSEAP is a sensitive, non-immunogenic reporter gene for preclinical mouse studies.     

Longitudinal river zonation in the tropics: examples of fish and caddisflies from the endorheic Awash River,Ethiopia

Specific concepts of fluvial ecology are well studied in riverine ecosystems of the temperate zone but poorly investigated in the Afrotropical region. Hence, we examined the longitudinal zonation of fish and adult caddisfly (Trichoptera) assemblages in the endorheic Awash River (1,250 km in length), Ethiopia. We expected that species assemblages are structured along environmental gradients, reflecting the pattern of large-scale freshwater ecoregions. We applied multivariate statistical methods to test for differences in spatial species assemblage structure and identified characteristic taxa of the observed biocoenoses by indicator species analyses. Fish and caddisfly assemblages were clustered into highland and lowland communities, following the freshwater ecoregions, but separated by an ecotone with highest biodiversity. Moreover, the caddisfly results suggest separating the heterogeneous highlands into a forested and a deforested zone. Surprisingly, the Awash drainage is rather species-poor: only 11 fish (1 endemic, 2 introduced) and 28 caddisfly species (8 new records for Ethiopia) were recorded from the mainstem and its major tributaries. Nevertheless, specialized species characterize the highland forests, whereas the lowlands primarily host geographically widely distributed species. This study showed that a combined approach of fish and caddisflies is a suitable method for assessing regional characteristics of fluvial ecosystems in the tropics.

     

Involvement of a carboxylated lysine in UV damage endonuclease

UV damage endonuclease is a DNA repair enzyme that can both recognize damage such as UV lesions and introduce a nick directly 5′ to them. Recently, the crystal structure of the enzyme from Thermus thermophilus was solved. In the electron density map of this structure, unexplained density near the active site was observed at the tip of Lys229. Based on this finding, it was proposed that Lys229 is post‐translationally modified. In this article, we give evidence that this modification is a carboxyl group. By combining activity assays and X‐ray crystallography on several point mutants, we show that the carboxyl group assists in metal binding required for catalysis by donating negative charge to the metal‐coordinating residue His231. Moreover, functional and structural analysis of the K229R mutant reveals that if His231 shifts away, an increased activity results on both damaged and undamaged DNA. Taken together, the results show that T. thermophilus ultraviolet damage endonuclease is carboxylated and the modified lysine is required for proper catalysis and preventing increased incision of undamaged DNA.     

End-point constraints in aiming movements: effects of approach angle and speed

 The present study focuses on two trajectory-formation models of point-to-point aiming movements, viz., the minimum-jerk and the minimum torque-change model. To date, few studies on minimum-jerk and minimum torque-change trajectories have incorporated self- or externally imposed end-point constraints, such as the direction and velocity with which a target area is approached. To investigate which model accounts best for the effects on movement trajectories of such – in many circumstances – realistic end-point constraints, we adjusted both the minimum-jerk and the minimum torque-change model so that they could generate trajectories of which the final part has a specific direction and speed. The adjusted models yield realistic trajectories with a high curvature near movement completion. Comparison of simulated and measured movement trajectories show that pointing movements that are constrained with respect to final movement direction and speed can be described in terms of minimization of joint-torque changes. Received: 7 July 1999 / Accepted in revised form: 8 January 2001     

Parental origin of chromosome abnormalities in spontaneous abortions

Summary Tissue cultures were initiated from 130 spontaneous abortion specimens and 81 were successfully karyotyped. Chromosome abnormalities were found in 50 cases: 12 with XO, 27 with trisomy, 6 with triploidy, 1 with tetraploidy and 4 others. The parental origin was determined in 11 cases of trisomy for an acrocentric chromosome. Two cases were uninformative while 9 non-disjunctions were determined and occurred during meiosis I: 7 were maternal and 2 paternal (both with trisomy 21). Three out of 7 cases with trisomy 16 were informative and resulted from a divisional error during the first meiotic division in the mother. All cases of triploidy were informative. They resulted from non-reduction during meiosis I in the mother (2) or dispermy (4).