Determination of PCR efficiency in chelex-100 purified clinical samples and comparison of real-time quantitative PCR and conventional PCR for detection of Chlamydia pneumoniae

Background  

Chlamydia pneumoniae infection has been detected by serological methods, but PCR is gaining more interest. A number of different PCR assays have been developed and some are used in combination with serology for diagnosis. Real-time PCR could be an attractive new PCR method; therefore it must be evaluated and compared to conventional PCR methods.     

First record of juvenile northern wolffish <Emphasis Type="Italic">Anarhichas denticulatus</Emphasis> Krøyer, 1845, on the continental slope of Northeast Greenland

The first record of the juvenile northern wolffish Anarhichas denticulatus was reported 570 km away from the nearest border of the species natural habitat near the island of Jan Mayen, on the continental slope of Northeast Greenland. Also original and published data on the occurrence of juvenile northern wolffish in the European Arctic and North Atlantic are presented.     

Androgen levels and components of aggressive behavior in men

Serum concentrations of testosterone (Tser), 5 alpha-dihydrotestosterone (DHT), and free testosterone (Tsal) in saliva were determined in 117 healthy young men between the ages of 20 and 30. A battery of standardized tests and projective techniques were administered simultaneously in order to measure various components of aggression, including sexual aggressiveness. All three androgens show reliable positive correlations with self-ratings of spontaneous aggression. Dominance exhibits a positive, statistically significant correlation to Tser and to DHT. In addition, DHT is negatively related to the scale restraint of aggression. These results support previous findings about Tser and point to the importance of other androgens--especially DHT--for this aspect of endocrine-affect relationships. Interest in sexual aggression yielded no significant results for Tser and DHT (Tsal shows a low positive correlation). The ratio DHT/Tser, however, correlates significantly with this component of aggression.     

PCR-based identification of microcystin-producing genotypes of different cyanobacterial genera

Microcystins are harmful hepatotoxins produced by many, but not all strains of the cyanobacterial genera Anabaena, Microcystis, Anabaena, Planktothrix, and Nostoc. Waterbodies have to be monitored for the mass development of toxic cyanobacteria; however, because of the close genetic relationship of microcystin-producing and non-producing strains within a genus, identification of microcystin-producers by morphological criteria is not possible. The genomes of microcystin-producing cells contain mcy genes coding for the microcystin synthetase complex. Based on the sequence information of mcy genes from Microcystis and Planktothrix, a primer pair for PCR amplification of a mcyA gene fragment was designed. PCR with this primer pair is a powerful means to identify microcystin-producing strains of the genera Anabaena, Microcystis, and Planktothrix. Moreover, subsequent RFLP analysis of the PCR products generated genus-specific fragments and allowed the genus of the toxin producer to be identified. The assay can be used with DNA from field samples.Abbreviations RFLP Restriction fragment length polymorphism - MALDI-TOF Matrix-assisted laser desorption/ionization-time of flight spectrometry - HPLC High performance liquid chromatography     

Interactions of neural glycosaminoglycans and proteoglycans with protein ligands: assessment of selectivity, heterogeneity and the participation of core proteins in binding

The method of affinity coelectrophoresis was used to study the binding of nine representative glycosaminoglycan (GAG)-binding proteins, all thought to play roles in nervous system development, to GAGs and proteoglycans isolated from developing rat brain. Binding to heparin and non-neural heparan and chondroitin sulfates was also measured. All nine proteins-laminin-1, fibronectin, thrombospondin-1, NCAM, L1, protease nexin-1, urokinase plasminogen activator, thrombin, and fibroblast growth factor-2-bound brain heparan sulfate less strongly than heparin, but the degree of difference in affinity varied considerably. Protease nexin-1 bound brain heparan sulfate only 1.8- fold less tightly than heparin (Kdvalues of 35 vs. 20 nM, respectively), whereas NCAM and L1 bound heparin well (Kd approximately 140 nM) but failed to bind detectably to brain heparan sulfate (Kd>3 microM). Four proteins bound brain chondroitin sulfate, with affinities equal to or a few fold stronger than the same proteins displayed toward cartilage chondroitin sulfate. Overall, the highest affinities were observed with intact heparan sulfate proteoglycans: laminin-1's affinities for the proteoglycans cerebroglycan (glypican-2), glypican-1 and syndecan-3 were 300- to 1800-fold stronger than its affinity for brain heparan sulfate. In contrast, the affinities of fibroblast growth factor-2 for cerebroglycan and for brain heparan sulfate were similar. Interestingly, partial proteolysis of cerebroglycan resulted in a >400- fold loss of laminin affinity. These data support the views that (1) GAG-binding proteins can be differentially sensitive to variations in GAG structure, and (2) core proteins can have dramatic, ligand-specific influences on protein-proteoglycan interactions.      

Requirements for carotenoids in fish diets

Carotenoids are among the most widespread and important pigment classes in living organisms, and in marine animals, astaxanthin is the most commonly occurring red carotenoid. Carotenoids are vitamin A precursors and are fundamental in photosynthesis and light protection in plants. Increasing attention has been drawn to a possible light protection, cancer prevention and immune enhancement by carotenoids in mammals. Reported functions in fish range from a general enhancement of performance to specific functions in reproduction and metabolism. In this paper, we show that astaxanthin is essential for growth and survival of fish and crawfish, and discuss this fact in relation to the use of purified and semipurified diets in nutritional studies. The similarity in action of astaxanthin and canthaxanthin compared to α-tocopherol (vitamin E) and retinol (vitamin A) suggests that these two carotenoids should be listed among the fat soluble vitamins.     

Free acetate production by rat hepatocytes during peroxisomal fatty acid and dicarboxylic acid oxidation

The fate of the acetyl-CoA units released during peroxisomal fatty acid oxidation was studied in isolated hepatocytes from normal and peroxisome-proliferated rats. Ketogenesis and hydrogen peroxide generation were employed as indicators of mitochondrial and peroxisomal fatty acid oxidation, respectively. Butyric and hexanoic acids were employed as mitochondrial substrates, 1, omega-dicarboxylic acids as predominantly peroxisomal substrates, and lauric acid as a substrate for both mitochondria and peroxisomes. Ketogenesis from dicarboxylic acids was either absent or very low in normal and peroxisome-proliferated hepatocytes, but free acetate release was detected at rates that could account for all the acetyl-CoA produced in peroxisomes by dicarboxylic and also by monocarboxylic acids. Mitochondrial fatty acid oxidation also led to free acetate generation but at low rates relative to ketogenesis. The origin of the acetate released was confirmed employing [1-14C]dodecanedioic acid. Thus, the activity of peroxisomes might contribute significantly to the free acetate generation known to occur during fatty acid oxidation in rats and possibly also in humans.     

West Nile virus: pending crisis for greater sage-grouse

Scientists have feared that emerging infectious diseases could complicate efforts to conserve rare and endangered species, but quantifying impacts has proven difficult until now. We report unexpected impacts of West Nile virus (WNv) on radio‐marked greater sage‐grouse (Centrocercus urophasianus), a species that has declined 45–80% and is endangered in Canada and under current consideration for federal listing in the US. We show that WNv reduced late‐summer survival an average of 25% in four radio‐marked populations in the western US and Canada. Serum from 112 sage‐grouse collected after the outbreak show that none had antibodies, suggesting that they lack resistance. The spread of WNv represents a significant new stressor on sage‐grouse and probably other at‐risk species. While managing habitat might lessen its impact on sage‐grouse populations, WNv has left wildlife and public health officials scrambling to address surface water and vector control issues in western North America.     

Fast calculation of the quartet distance between trees of arbitrary degrees

Background  

A number of algorithms have been developed for calculating the quartet distance between two evolutionary trees on the same set of species. The quartet distance is the number of quartets – sub-trees induced by four leaves – that differs between the trees. Mostly, these algorithms are restricted to work on binary trees, but recently we have developed algorithms that work on trees of arbitrary degree.