ABSTRACT
The objective was to develop a high-throughput method of identifying sex in both Coturnix chinensis and Gallus gallus, which would be useful for biomedical research and hatcheries. Because chromo-helicase-DNA binding protein (CHD)-based Griffiths P2/P8 primers do not produce polymerase chain reaction (PCR) products with distinguishable sex-specific curves in melting curve analysis (MCA), these primers are unsuitable for high throughput application in either species. Conserved regions were identified by basic local alignment search tool (BLAST) analyses of cloned CHD-Z and CHD-W genes of C. chinensis. Based on sequence alignment, a female-specific CHD-W primer (W-cot-F1) and a female/male (or CHD-W/CHD-Z)-common primer (ZW-cot-F1) were redesigned for use in combination with the Griffiths P2 primer for MCA-based PCR reaction. In C. chinensis and G. gallus, W-cot-F1/P2 and ZW-cot-F1/P2 had amplicon lengths of 315/318 and 114 base pairs and melting temperatures (Tm) of approximately 79.5 °C to 80 °C and approximately 78.5 °C to 79°C, respectively. Thus, MCA distinguished sex based on two distinct Tm peaks in females versus only one Tm peak in males. The MCA-based real-time PCR combined with the proposed primer redesign provided a high-throughput method of identifying sex in C. chinensis and G. gallus.
Subject(s)
Chickens/physiology , Quail/physiology , Sex Determination Analysis/veterinary , Animals , Avian Proteins/genetics , Avian Proteins/metabolism , Base Sequence , Cloning, Molecular , DNA Primers , DNA-Binding Proteins , Female , Gene Expression Regulation , Male , Nucleic Acid Denaturation , Sequence Analysis, DNA , Sex Determination Analysis/methodsABSTRACT
The objective of this study was to test the hypothesis that genders of Accipitridae species, with the same or similar sequences to our previously proposed Spilornis cheela hoya (S. c. hoya) chromo-helicase-DNA binding protein (CHD)-W-specific and CHD-ZW-common TaqMan probes, can be successfully determined. Eight species of Accipitridae with known genders were collected. After PCR, TA cloning, sequencing, and alignment analyses, sequence length differences of Griffiths P2/P8 PCR amplicons between CHD-Z and CHD-W genes ranged from 2 to 19 bp for these Accipitridae species, and they were unsolved in 3% agarose gel. Using our previous proposed S. c. hoya TaqMan probes, the genders of Circaetus gallicus, completely homologous to the sequences for these CHD probes, were successfully identified. With one nucleotide difference to S. c. hoya CHD-W-specific probe, gender identification of Accipiter gularis, Accipiter soloensis, Accipiter trivirgatus, Accipiter virgatus, and Butastur indicus were validated. With two nucleotide differences in the CHD-W-specific probe and one nucleotide difference in the CHD-ZW-common probe, Pernis ptilorhyncus also performed well for gender identification. In conclusion, the S. c. hoyaCHD probes, coupled with the Griffiths P2/P8 primers, were validated to provide accurate and high-throughput gender identification for many Accipitridae species.
Subject(s)
Falconiformes/genetics , Sex Determination Processes , Animals , Base Sequence , DNA Primers , Female , Male , Molecular Probes , Molecular Sequence Data , Polymerase Chain Reaction/methods , Sequence Alignment , Sequence Analysis, DNAABSTRACT
Extra-corporeal life support (ECLS) has been applied successfully to congenital respiratory defects but less optimally to acquired pulmonary failure. We extended this support to certain extreme complexities of patients with acute respiratory distress. From January 2003 to June 2005, 16 (nine men and seven women) patients refractory to ventilator support were treated with ECLS. Their median age was 32.4 years (1.5-70). The triggering events were pulmonary haemorrhage (n = 4), pneumonia (n = 7), aspiration (n = 2) and pancreatitis (n = 3). The indications for support were hypoxaemia in 13 and hypercapnia in three patients. Ten (63%) met the criteria of fast entry. Thirteen (81%) received veno-venous (V-V) mode support and the other three received veno-arterial mode support initially, but then converted to V-V mode after sufficient oxygenation stabilised haemodynamics. Initial pump flow was maximised to improve (mean 3250 +/- 1615 ml/min) to improve the oxygenation. Four patients with active pulmonary haemorrhage were heparin free in the first 12-24 h of support without complications. Excluding one prematurely terminated patient because of brain permanent damage, the duration of support was 162 +/- 95 h (67-363). Eleven (69%) weaned successfully from ECLS and 10 (63%) discharged and regained normal pulmonary performance in a median of 26.8 months follow-up. Pulmonary support using ECLS was feasible in selected patients with acute respiratory distress. Modification of guidelines for liberal use, early deployment before secondary organ damage and prevention of complications during support were the key to final success.
Subject(s)
Extracorporeal Membrane Oxygenation/methods , Respiratory Distress Syndrome/therapy , Adolescent , Adult , Aged , Algorithms , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Patient Selection , Respiratory Distress Syndrome/etiology , Treatment Outcome , Ventilator Weaning/methodsABSTRACT
Mycotic aneurysms are an important cause of morbidity and mortality in endocarditis despite advanced antibiotic therapy. Visceral artery aneurysms are uncommon and usually remain clinically silent until rupture. We now report a case of successful surgical treatment of a superior mesenteric mycotic aneurysm of the superior mesenteric artery, followed by a review of pertinent clinical information.
Subject(s)
Aneurysm, False/etiology , Aortic Valve , Endocarditis, Bacterial/complications , Heart Valve Diseases/complications , Mesenteric Artery, Superior , Adult , Aneurysm, False/surgery , Aneurysm, Infected/etiology , Aneurysm, Infected/surgery , Humans , Male , Mesenteric Artery, Superior/surgeryABSTRACT
An antifungal metabolite, bacereutin, was isolated from culture filtrate of Bacillus cereus CHU 130. The bacterium was isolated from soils collected in Changhwa County, Taiwan, and was grown in soybean meal-mannitol broth for production of the antibiotic metabolite. The antibiotic metabolite was isolated by adsorption column chromatography of Amberite XAD-2 and was purified by passing through the chromatographic columns packed with Dowex 50W-X8, Sephadex LH 20 and Biogel P-2. The antibiotic metabolite was soluble in water and 87% acetone, and was slightly soluble in methanol, but was not dissolved in n-propanol, n-butanol, acetone, benzene and ethyl acetate. The antibiotic metabolite was a heat-stable and ninhydrin-positive substance. The antibiotic activities of bacereutin were tested by means of the agar-diffusion plate method. The antibiotic metabolite inhibited the growth of Saccharomyces cerevisiae CHU 1, Paecilomyces variotii CHU 6, Rhizomucor miehei CHU 40 and Fusarium oxysportum CHU 98. Bacereutin was a ninhydrin-positive antifungal antibiotic.
