Characterization of polymorphic microsatellite markers for the Carnaby's cockatoo (Calyptorhynchus latirostris) and related black cockatoo species

Microsatellite loci were isolated from Carnaby's black cockatoo (Calyptorhynchus latirostris: Aves), a highly valued, endangered, and endemic species of bird from Western Australia. This study describes three dinucleotide and one tetranucleotide microsatellite loci for which the primers produced clear and polymorphic amplification patterns with between two and nine alleles and moderate levels of variability. Two additional dinucleotide markers which were monomorphic in the Carnaby's cockatoo were able to amplify and were polymorphic in two other species of black cockatoo, greatly increasing the utility of these markers.     

Influence of red pepper and capsaicin on body composition and lipogenesis in rats

Inclusion of red pepper or its active principle ‘capsaicin’ in the diet led to a lowering of total lipids, particularly triglycerides in the liver. The total body fat was lowered in animals fed red pepper or capsaicin but not in animals fed paprika powder which had negligible capsaicin content. Hyperlipogensis and hypertriglyceridemia caused by fructose feeding were significantly were decreased in capsaicin-fed animals. Activities of the key lipogenic enzymes were reduced as reflected by decreased lipogenesis.     

Circadian rhythm of serum and tissue lipids in fed and fasted rats

The oscillations of the free fatty acid concentration in the serum and white (epididymal) adipose tissue, of triglycerides in the serum and liver, of total serum, liver and adrenal cholesterol and of serum phospholipids were studied at 3-hour intervals for a period of 24 hours in fed male Wistar rats and in animals fasted for 24 hours (both adapted to an illumination regimen of 12 hours' light and 12 hours' darkness. The rhythm--studied by means of the cosinor analysis--was present in most of the given parameters; it was not recorded in the liver triglycerides and serum phospholipids of fasted rats and in the adrenal cholesterol of fed animals. Apart from the circadian rhythm, many parameters distinctly displayed an ultradian rhythm, mainly an approximately 12-hour period. In general, one day's starvation did not significantly affect the course of the circadian oscillations of the given indicators of rat lipid metabolism.     

Mutation of residues 423 (Met/Ile), 444 (Thr/Met), and 506 (Asn/Ser) confer cholesteryl esterase activity on rat lung carboxylesterase. Ser-506 is required for activation by cAMP-dependent protein kinase.

Site-directed mutagenesis is used to identify amino acid residues that dictate reported differences in substrate specificity between rat hepatic neutral cytosolic cholesteryl ester hydrolase (hncCEH) and rat lung carboxylesterase (LCE), proteins differing by only 4 residues in their primary sequences. Beginning with LCE, the substitution Met(423) --> Ile(423) alone or in combination with other mutations increased activity with p-nitrophenylcaprylate (PNPC) relative to more hydrophilic p-nitrophenylacetate (PNPA), typical of hncCEH. The substitution Thr(444) --> Met(444) was necessary but not sufficient for expression of cholesteryl esterase activity in COS-7 cells. The substitution Asn(506) --> Ser(506), creating a potential phosphorylation site, uniformly increased activity with both PNPA and PNPC, was necessary but not sufficient for expression of cholesteryl esterase activity and conferred susceptibility to activation by cAMP-dependent protein kinase, a property of hncCEH. The 3 mutations in combination were necessary and sufficient for expression of cholesteryl esterase activity by the mutated LCE. The substitution Gln(186) --> Arg(186) selectively reduced esterase activity with PNPA and PNPC but was not required for cholesteryl esterase activity. Homology modeling from x-ray structures of acetylcholinesterases is used to propose three-dimensional models for hncCEH and LCE that provide insight into the effects of these mutations on substrate specificity.     

Application of avidin-ferritin and peroxidase as contrasting electron-dense markers for simultaneous electron microscopic immunocytochemical labelling of glutamic acid decarboxylase and tyrosine hydroxylase in the rat arcuate nucleus

Summary A pre-embedding immunostaining procedure was developed using ferritin and peroxidase to enable simultaneous electron microscopic localization of two antigens in the same tissue section. This method was used to study the anatomic relationship between glutamic acid decarboxylase (GAD) immunoreactive axons and tyrosine hydroxylase (TH) — containing neurons of the rat arcuate nucleus. The findings provide ultrastructural evidence that GAD-immunoreactive terminals establish symmetric (Gray II) synapses on TH-reactive neurons.     

Stimulation of tissue plasminogen activator production from epithelial cell lines

The aim of this study was to investigate the possibility of enhancing the yield of tissue plasminogen activator (tPA) from two epithelial cell lines of normal (non-malignant) derivation grown in tissue culture. The three agents used in this investigation were chosen because of their proven enhancing effect on analogous cells or products. The anabolic hormone stanozolol was found to have no significant stimulatory effect on these cell lines. A phorbol acetate (12-O-tetradecanoylphorbol 13-acetate) caused a twofold enhancement in tPA yield but the most significant results were obtained with 5-azacytidine. This agent increased the yield by up to fourfold in small stationary cultures and threefold in large-scale microcarrier cultures. A combination of azacytidine and phorbol acetate did not have an additive effect on total yield but did alter the kinetics of tPA expression with time. Indications were that the maximum yield with these types of potentiating agents was achieved as it could not be increased by using a combination of two different agents.     

Energy from vascular plant wastewater treatment systems

Water hyacinth (Eichhornia crassipes), duckweed (Spirodela sp. andLemna sp.), water pennywort (Hydrocotyle ranunculoides), and kudzu (Pueraria lobata) were anaerobically fermented using an anaerobic filter technique that reduced the total digestion time from 90 d to an average of 23 d and produced 0.14-0.22 m³ CH₄/kg (dry weight) (2.3-3.6 ft³/lb) from mature filters for the 3 aquatic species. Kudzu required an average digestion time of 33 d and produced an average of 0.21 m³ CH₄/kg (dry weight) (3.4 ft³/lb). The anaerobic filter provided a large surface area for the anaerobic bacteria to establish and maintain an optimal balance of facultative, acid-forming, and methane-producing bacteria. Consequently the efficiency of the process was greatly improved over prior batch fermentations.