80 research outputs found

    Development of an artificial weaning diet for the South African abalone, Haliotis midae (Haliotidae: Gastropoda)

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    An adequate supply of diatoms during the weaning stage (generally 5 - 10 mm shell length (SL)) is one of the primary constraints to the commercial culture of the South African abalone, Haliotis midae. Because of the seriousness of the problem, a project aimed at the development of an artificial weaning diet was initiated. Initially, the chemical composition (proximate composition, amino acid, fatty acid and mineral element profile) of juvenile H. midae was analyzed, as a general lack of such information was identified in a review. Due to the lack of knowledge on the nutritional requirements of H. midae, the formulation of the weaning diet was based on the essential amino acid (EAA) pattern of the shucked tissue, and the known nutrient requirements of haliotids. Subsequently, a water stable gel and pellet form of the diet were developed. The best water stability of a gel was obtained with a 1:3 agar/gelatine mixture which retained 70.7 ± 2.7 % of its dry weight after 24 h. Starch based pellets, however, retained 89.0 ± 0.6 % of their dry weight after 24 h. In a comparative growth trial, pellets produced a significantly better increase in SL and weight than gels after only 15 days. This was probably due to the better water stability of pellets, which resulted in a better nutritional quality than in gels. The feeding behaviour on both forms of the diet did not differ. Activity patterns were exclusively nocturnal and feeding frequency was consistently low. The percentage composition of the pelleted weaning diet, on a dry weight basis, was 5 % casein, 15 % gelatine, 15 % fish meal, 10 % Spirulina spp., 2.5 % fish oil, 2.5 % sunflower oil, 21.0 % dextrin, 23.0 % starch, 4.0 % of a mineral and 2.0 % of a vitamin mixture. The correlation coefficient between the EAA pattern of H. midae and the dietary EAA pattern was r⁷= 0.8989. Pellets were fed to juveniles in a 30 day growth trial to study the effect of photoperiod (12, 16, 20 and 23 hours of darkness) on growth and general nutritional parameters. A comparative experiment feeding diatoms was conducted under a 12hL: 12hD light regime at the same time. The SL and weight of the juveniles did not increase significantly with an increase in hours of darkness. The growth of juveniles fed on pellets did not differ significantly from those fed on diatoms. Percentage feed consumption (PFC), percentage feeding rate (PFR), feed conversion ratio (FCR), protein efficiency ratio (PER) and percentage protein deposited (PPD) were determined for the animals fed on pellets. None of the parameters were significantly affected by photoperiod. However, there were trends in that PFC increased with longer periods of darkness, while PPD decreased. The FCRs (0.44 ± 0.04 to 0.60 ± 0.19) and PERs (5.06 ± 1.74 to 6.64 ± 0.77) indicated that juveniles used the feed, and in particular the protein, very efficiently. Photoperiod did not have an effect on the specific activity of the digestive enzymes amylase, protease and lipase. The specific activity of amylase in the juveniles fed on diatoms was significantly higher than in the pellet fed groups. This was surprising as the main carbohydrate of diatoms is the ß-(l-3) glucan chrysolaminarin, and not starch, a ß-(l-4) glucan. Protease specific activity, on the other hand, was significantly higher in the pellet fed groups, indicating an ability to adapt to the high protein content in the artificial diet (35.48 %), compared to diatoms which had a protein content of 5 %. The specific activity of lipase did not differ significantly between groups, probably because of a similar lipid concentration (5 - 10 %) in diatoms and pellets. Finally, the effect of stocking density, ranging from 1250 to 10,000 juveniles/m2, on the growth of juveniles was evaluated. A model of hatchery productivity was developed based on this investigation. Hatchery productivity was defined as the number of juveniles per unit space reared through to the grow-out stage per unit time. The model predicted that maximum productivity would be achieved at a stocking density of 10,000 juveniles/m2. The results have shown that H. midae can be successfully weaned on an artificial diet, as the growth on the diet was not significantly different to growth obtained on diatoms. Long-term growth trials are needed to confirm these results. The importance of standardized experiments on the nutritional requirements and digestibility of abalone was emphasized. The importance of improved artificial diets, optimal culture conditions, as well as the application of biotechnological techniques to further abalone aquaculture was highlighted

    Das Photoelektronen- und Elektronenspektrum des 1,3,5-Tri-tert-butylpentalens. Hinweis auf Bindungsalternanz im Grundzustand

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    Das Photoelektronenspektrum (He(I)) und das Absorptionsspektrum (UV-VIS) von 1,3,5-Tri-tert-butylpentalen (1) werden diskutiert. Die Zuordnung der ersten vier Banden erfolgt aufgrund eines Vergleichs mit den berechneten Orbitalenergien bzw. ÜbergĂ€ngen. Beim Absorptionsspektrum von 1 ist nur dann eine gute Übereinstimmung zwischen Experiment und Rechnung zu erzielen, wenn Bindungsalternanz bei 1 angenommen wird

    Bioassays to Monitor Taspase1 Function for the Identification of Pharmacogenetic Inhibitors

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    Background: Threonine Aspartase 1 (Taspase1) mediates cleavage of the mixed lineage leukemia (MLL) protein and leukemia provoking MLL-fusions. In contrast to other proteases, the understanding of Taspase1's (patho)biological relevance and function is limited, since neither small molecule inhibitors nor cell based functional assays for Taspase1 are currently available. Methodology/Findings: Efficient cell-based assays to probe Taspase1 function in vivo are presented here. These are composed of glutathione S-transferase, autofluorescent protein variants, Taspase1 cleavage sites and rational combinations of nuclear import and export signals. The biosensors localize predominantly to the cytoplasm, whereas expression of biologically active Taspase1 but not of inactive Taspase1 mutants or of the protease Caspase3 triggers their proteolytic cleavage and nuclear accumulation. Compared to in vitro assays using recombinant components the in vivo assay was highly efficient. Employing an optimized nuclear translocation algorithm, the triple-color assay could be adapted to a high-throughput microscopy platform (Z'factor = 0.63). Automated high-content data analysis was used to screen a focused compound library, selected by an in silico pharmacophor screening approach, as well as a collection of fungal extracts. Screening identified two compounds, N-[2-[(4-amino-6-oxo-3H-pyrimidin-2-yl)sulfanyl]ethyl]benzenesulfonamideand 2-benzyltriazole-4,5-dicarboxylic acid, which partially inhibited Taspase1 cleavage in living cells. Additionally, the assay was exploited to probe endogenous Taspase1 in solid tumor cell models and to identify an improved consensus sequence for efficient Taspase1 cleavage. This allowed the in silico identification of novel putative Taspase1 targets. Those include the FERM Domain-Containing Protein 4B, the Tyrosine-Protein Phosphatase Zeta, and DNA Polymerase Zeta. Cleavage site recognition and proteolytic processing of these substrates were verified in the context of the biosensor. Conclusions: The assay not only allows to genetically probe Taspase1 structure function in vivo, but is also applicable for high-content screening to identify Taspase1 inhibitors. Such tools will provide novel insights into Taspase1's function and its potential therapeutic relevance

    Preclinical good laboratory practice-compliant safety study to evaluate biodistribution and tumorigenicity of a cartilage advanced therapy medicinal product (ATMP)

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    Background: The clinical development of advanced therapy medicinal products (ATMPs), a new class of drugs, requires initial safety studies that deviate from standard non-clinical safety protocols. The study provides a strategy to address the safety aspects of biodistribution and tumorigenicity of ATMPs under good laboratory practice (GLP) conditions avoiding cell product manipulation. Moreover, the strategy was applied on a human ATMP for cartilage repair

    Evaluation of the preventive capacities of a topically applied azithromycin formulation against Lyme borreliosis in a murine model

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    Objectives: Systemic antibiotic treatment of Lyme borreliosis is effective during the early stages of the infection, while chronic manifestations of the disease may remain refractory and difficult to treat. This study was carried out in order to evaluate the potential of topically applied azithromycin to eliminate the spirochaetal organisms in the skin of the freshly bitten host and thereby prevent Lyme borreliosis. Methods: Laboratory mice were challenged with Borrelia burgdorferi sensu stricto by needle inoculation or via infected ticks as vectors. Then, an azithromycin-containing formulation was applied once daily to the sites of exposure for three consecutive days. In the case of needle inoculation, a 5% azithromycin formulation was applied starting 1 h, 3 days and 5 days after infection. In the case of tick exposure, 4%, 10% and 20% azithromycin formulations were applied, starting directly after the detachment of the engorged ticks. Subsequently, the infection status of the mice was determined. Results: Concentrations of azithromycin in murine skin were >3800-fold higher than the published minimal inhibitory concentration for B. burgdorferi as soon as 3 h after the first application. After needle inoculation, spirochaetes were not detectable in all infected mice after treatment, if the first application started 1 h or even after 3 days post-infection. Furthermore, no borrelial organisms were detected after topical treatment when ticks were used for spirochaete inoculation. Conclusions: Our data indicate that topical treatment with a formulation containing azithromycin is a promising approach to prevent Lyme borreliosis shortly after a tick bite

    Advanced electron cyclotron heating and current drive experiments on the stellarator Wendelstein 7-X

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    During the first operational phase (OP 1.1) of Wendelstein 7-X (W7-X) electron cyclotron resonance heating (ECRH) was the exclusive heating method and provided plasma start-up, wall conditioning, heating and current drive. Six gyrotrons were commissioned for OP1.1 and used in parallel for plasma operation with a power of up to 4.3 MW. During standard X2-heating the spatially localized power deposition with high power density allowed controlling the radial profiles of the electron temperature and the rotational transform. Even though W7-X was not fully equipped with first wall tiles and operated with a graphite limiter instead of a divertor, electron densities of n e > 3·1019 m-3 could be achieved at electron temperatures of several keV and ion temperatures above 2 keV. These plasma parameters allowed the first demonstration of a multipath O2-heating scenario, which is envisaged for safe operation near the X-cutoff-density of 1.2·1020 m-3 after full commissioning of the ECRH system in the next operation phase OP1.2

    The antiapoptotic gene survivin is highly expressed in human chondrosarcoma and promotes drug resistance in chondrosarcoma cells in vitro

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    Background Chondrosarcoma is virtually resistant to chemotherapy and radiation therapy. Survivin, the smallest member of the inhibitor of apoptosis protein family, is a critical factor for tumor progression and resistance to conventional therapeutic approaches in a wide range of malignancies. However, the role of survivin in chondrosarcoma has not been well studied. We examined the importance of survivin gene expression in chondrosarcoma and analysed its influences on proliferation, apoptosis and resistance to chemotherapy in vitro. Methods Resected chondrosarcoma specimens from which paraffin-embedded tissues could be extracted were available from 12 patients. In vitro experiments were performed in human chondrosarcoma cell lines SW1353 and Hs819.T. Immunohistochemistry, immunoblot, quantitative PCR, RNA interference, gene-overexpression and analyses of cell proliferation and apoptosis were performed. Results Expression of survivin protein was detected in all chondrosarcoma specimens analyzed, while undetectable in adult human cartilage. RNA interference targeting survivin resulted in a G2/M-arrest of the cell cycle and led to increased rates of apoptosis in chondrosarcoma cells in vitro. Overexpression of survivin resulted in pronounced resistance to doxorubicin treatment. Conclusions These findings indicate that survivin plays a role in the pathogenesis and pronounced chemoresistance of high grade chondrosarcoma. Survivin antagonizing therapeutic strategies may lead to new treatment options in unresectable and metastasized chondrosarcoma
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