Parasite diversity as an indicator of environmental change? An example from tropical grouper (Epinephelus fuscoguttatus) mariculture in Indonesia.

Fish parasites are used to monitor long-term change in finfish grouper mariculture in Indonesia. A total of 210 Epinephelus fuscoguttatus were sampled in six consecutive years between 2003/04 and 2008/09 and examined for parasites. The fish were obtained from floating net cages of a commercially run mariculture facility that opened in 2001. The fauna was species rich, consisting of ten ecto- and 18 endoparasite species. The ectoparasite diversity and composition was relatively stable, with the monogeneans Pseudorhabdosynochus spp. (83-100% prevalence, Berger-Parker Index of 0·82-0·97) being the predominant taxon. Tetraphyllidean larvae Scolex pleuronectis and the nematodes Terranova sp. and Raphidascaris sp. 1 were highly abundant in 2003/04-2005/06 (max. prevalence S. pleuronectis 40%, Terranova sp. 57%, Raphidascaris sp. 1 100%), and drastically reduced until 2008/09. These parasites together with the prevalence of Trichodina spp., ecto-/endoparasite ratio and endoparasite diversity illustrate a significant change in holding conditions over the years. This can be either referred to a definite change in management methods such as feed use and fish treatment, or a possible transition of a relatively undisturbed marine environment into a more affected habitat. By visualizing all parameters within a single diagram, we demonstrate that fish parasites are useful bioindicators to monitor long-term change in Indonesian grouper mariculture. This also indicates that groupers can be used to monitor environmental change in the wild. Further taxonomic and systematic efforts in less sampled regions significantly contributes to this new application, supporting fish culture and environmental impact monitoring also in other tropical marine habitats.

Off-label use in Germany - a current appraisal of gynaecologic university departments.

OBJECTIVE: The off-label use, referring to the applicability of pharmaceutical drugs beyond the submitted and from the Federal Institute for Drugs and Medical Devices (BfArM, Bundesamt für Arzneimittel und Medizinprodukte) certified and approved administration, is the subject of controversial discussions. The application can be considered in case of severe illness - if no therapeutic alternatives are available - or it exists as a founded perspective for achieving therapeutic success. METHODS: A latitudinal study for evaluating the application of off-label use supplements was performed at 43 German university and academic teaching hospitals. Five doctors at each hospital applied off-label pharmaceutical drugs and were called upon to share their personal experience to the application of those medications. RESULTS: 75 (35%) questionnaires were returned out of 22 (51%) medical centres with 215 contacted physicians. Off-label use was common for 65 (91%) of the physicians. Only 9% of them obviate the application of off-label drugs. About a half of the medication is related to application in obstetrics (54%) and in most cases on an every day basis. Uterotonics were the most commonly used off-label medications (34%). The main part of information about off-label use is obtained from personal information of colleagues (66%) and personal experience (58%). 34% of physicians think that off label use is risky. Interestingly, the view about off label use of medication varies considerably among physicians from various hospitals. CONCLUSIONS: The application of off-label pharmaceutical drugs in Germany seems to be a well established practice. More than 90% of participators of our trial use at least one medication outside the administration. This includes particularly prostaglandins, anti-hypertonic therapeutics and chemotherapeutics.

A new approach to visualize ecosystem health by using parasites.

A new approach is chosen to visualize ecosystem health by using parasite bioindicators in Segara Anakan Lagoon, a brackish water ecosystem at the southern Java coast, Indonesia. Three fish species (Mugil cephalus, Scatophagus argus, Epinephelus coioides) were collected in two different years and sampling sites and studied for ecto- and endoparasites. Additional data were taken for E. coioides from two further sites in Lampung Bay, Sumatra, and for E. fucoguttatus out of floating cages from a mariculture facility in the Thousand Islands, Jakarta Bay, North Java. The parasite fauna of fishes inside the lagoon was characterized by a high number of ecto- and a low number of endoparasites, the endoparasite diversity was relatively low and the prevalence of ectocommensalistic trichodinid ciliates was high. These parameters were chosen to indicate the biological conditions inside the lagoon. In E. coioides during rainy season, the prevalence of trichodinid ciliates was highest inside the lagoon (55%) compared with 27% in an open-net-cage mariculture and 5.7% in free-living specimens in Lampung Bay. The endoparasite diversity (Shannon-Wiener) was lowest in fish from Segara Anakan lagoon (0.66) compared with fish from an open-net-cage mariculture (0.71) and free-living specimens (1.39). Results for E. fuscoguttatus from the mariculture site in the Thousand Islands, a relatively undisturbed marine environment, demonstrated high parasite diversity (1.58) in the cultivated fish, a high number of endoparasites, and no trichodinids. A star graph is used to visualize the parasite composition for the different fishes, sampling sites, and conditions, using (1) the prevalence of trichodinid ciliates, (2) the ecto/endoparasite ratio and (3) the endoparasite diversity as bioindicators. The application of the star graph is suggested to be a suitable tool to visualize and monitor environmental health under high parasite biodiversity conditions within tropical ecosystems. It can also support a better communication to stake holders and decision makers in order to monitor environmental impact and change.

The effects of glucose, insulin and metformin on the order parameters of isolated red cell membranes. An electron paramagnetic resonance spectroscopic study.

Human red blood cell (RBC) membranes (RBC ghosts) were treated with glucose, insulin and metformin. The order parameters of RBC membranes were determined by 5- and 16-doxyl-stearic acid spin labels. Metabolic effects were excluded using an isolated system of RBC membranes. The membranes were incubated with glucose in physiological (5 mM), renal threshold (10 mM) and manifested diabetic (20 mM) concentrations for limited times. High concentrations of glucose (10, 20, 100 mM) increase the order parameters of RBC membranes significantly. Insulin by itself has a similar effect which is, however, not strictly concentration-dependent. By contrast, metformin at therapeutic concentrations (0.5 and 5.0 microM) decreases the order parameters. At 50 microM concentration the metformin effect is expressed less and recurs at 100 microM concentration. The effects are significant with 5-doxyl-stearic acid, but are not significant with the 16-doxyl derivative. When RBC membranes are co-incubated with 20 mM glucose and metformin at 0.5 and 5.0 microM concentrations the order parameters as determined by 5-doxyl-stearic acid remain normal (= control values). Higher concentrations of metformin (50 and 100 microM) cause an overshoot to very low order parameters. Insulin at 10, 100 and 200 mU/L does not influence significantly the effects of metformin. Addition of physiological amounts of bovine serum albumin does not abolish the effects of metformin. Metformin, at therapeutic concentrations (0.5 and 5.0 microM), maintains the normal fluidity at the polar interface of isolated RBC membranes by counterbalancing non-enzymatic glycosylation with 20 mM glucose in vitro.

[Automated detection of spleen volume by spiral CT scans using neural networks and "fuzzy logic"]. / Automatische Berechnung des Milzvolumens aus Spiral-CT-Daten mit Hilfe neuronaler Netze und "Fuzzy Logik".

PURPOSE: To assess spleen segmentation and volumentry in spiral CT scans with and without pathological changes of splenic tissue. METHODS: The image analysis software HYBRIKON is based on region growing, self-organized neural nets, and fuzzy-anatomic rules. The neural nets were trained with spiral CT data from 10 patients, not used in the following evaluation on spiral CT scans from 19 patients. An experienced radiologist verified the results. The true positive and false positive areas were compared in terms to the areas marked by the radiologist. The results were compared with a standard thresholding method. RESULTS: The neural nets achieved a higher accuracy than the thresholding method. Correlation coefficient of the fuzzy-neural nets: 0.99 (thresholding: 0.63). Mean true positive rate: 90% (thresholding: 75%), mean false positive rate: 5% (thresholding > 100%). Pitfalls were caused by accessory spleens, extreme changes in the morphology (tumors, metastases, cysts), and parasplenic masses. CONCLUSIONS: Self-organizing neural nets combined with fuzzy rules are ready for use in the automatic detection and volumetry of the spleen in spiral CT scans.

7-Deaza-2'-deoxyguanosine allows PCR and sequencing reactions from CpG islands.

CpG islands are GC rich sequences that are found in the promoters of many genes in higher eukaryotes. They contain a high frequency of CG dinucleotides, which are substrates for DNA methylases. Methylation leads to transcriptional silencing of promoters. Owing to their high GC content CpG islands exhibit strong base-base interactions, which lead to superstructures and consequently to regions with higher melting temperatures. Therefore, Taq polymerases (especially sequenases) fall off their templates, causing premature termination of the polymerase chain reaction (PCR) or sequencing reactions. The results from such reactions are thus insufficient for further analysis. Therefore, we have evaluated the use of 7-deaza-2'-deoxyguanosine for PCR amplification of the human p16(INK4A) promoter and sequencing of HUMARA exon 1 PCR products. Our results show that the addition of 7-deaza-2'-deoxyguanosine significantly improves results, particularly when small amounts of poor quality DNA are available as starting material.