Background Mice are well suited for modeling human congenital heart defects (CHD), given their four-chamber cardiac anatomy. However, mice with CHD invariably die prenatally/neonatally, causing CHD phenotypes to be missed. Therefore, we investigated the efficacy of noninvasive micro-computed tomography (micro-CT) to screen for CHD in stillborn/fetal mice. These studies were carried out using chemically mutagenized mice expected to be enriched for birth defects including CHD. Methods and Results Stillborn/fetal mice obtained from the breeding of N-ethyl-N-nitrosourea (ENU) mutagenized mice were formalin-fixed and stained with iodine, then micro-CT scanned. Those diagnosed with CHD and some CHD-negative pups were necropsied. A subset of these were further analyzed by histopathology to confirm the CHD/no-CHD diagnosis. Micro-CT scanning of 2105 fetal/newborn mice revealed an abundance of ventricular septal defects (VSD) (n=307). Overall, we observed an accuracy of 89.8% for VSD diagnosis. Outflow tract anomalies identified by micro-CT included double outlet right ventricle (n=36), transposition of the great arteries (n=14), and persistent truncus arteriosus (n=3). These were diagnosed with a 97.4% accuracy. Aortic arch anomalies also were readily detected with an overall 99.6% accuracy. This included right aortic arch (n=28) and coarctation/interrupted aortic arch (n=12). Also detected by micro-CT were atrioventricular septal defects (n=22), tricuspid hypoplasia/atresia (n=13), and coronary artery fistulas (n=16). They yielded accuracies of 98.9%, 100%, and 97.8% respectively. Conclusions Contrast enhanced micro-CT imaging in neonatal/fetal mice can reliably detect a wide spectrum of CHD. We conclude micro-CT imaging can be used for routine rapid assessments of structural heart defects in fetal/newborn mice.
Sequential optimization strategy was used to enhance the production of extracellular polygalacturonase by a newly isolated Aspergillus fumigatus R6 using rice bran as a substrate in solid state conditions. Three significant variables influencing the polygalacturonase production were identified as initial moisture level, temperature and incubation time (P < 0.0001). The model established by face-centered central composite design was significant (P < 0.05) with high R2 (0.98). The model validity was verified and the optimum conditions were at an initial moisture level of 49.6%, 33 °C and 129 h of incubation time with the maximum polygalacturonase activity of 565 U/g, resulted in 2.65 fold increase in polygalacturonase activity compare to the unoptimized conditions. Kenaf stem treated with A. fumigatus R6 polygalacturonase enzyme at 72 h produced high strength of kenaf bast fibers (287 MPa) with high Young's modulus (10404 MPa) and the color is in satisfactory.
Enzyme retting can be a viable alternative to water retting, which is the currently utilised method for extracting fibres from kenaf. The advantages of enzyme retting are its greater environmental friendliness, shorter retting time, and more controllable fibre quality. The objective of this study was to determine the efficacy of pectinase produced from locally isolated Aspergillus fumigatus R6 in kenaf retting. A. fumigatus R6 pectinase effectively separated the fibres from non-fibre components. Scanning electron micrographs showed that the surface of pectinase-treated kenaf bast fibres appeared to be smoother and finer. The degree of retting increased with incubation time. A retting time of 32 h produced goodquality kenaf bast fibres with high tensile strength (459 MPa). No significant differences were found between the tensile properties of kenaf bast fibres treated with A. fumigatus R6 pectinase-containing culture filtrate and other sources of commercial pectinase enzyme. Hence, it was concluded that A. fumigatus R6 pectinase was capable of retting kenaf effectively.
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