Surface bands on deepwater squalid dorsal-fin spines : an alternative method for ageing centroselachus crepidater

Bands on the external surface of the second dorsal-fin spine proved to be a novel method of estimating the age and growth of Centroselachus crepidater. Bands that followed the shape of the spine base were enhanced with an alizarin red derivative. Internal bands in spine cross sections were also examined. The number of both external and internal bands increased with animal size, although most spines had more external than internal bands. External bands were more reliable and were assumed to be annuli. The rate of band formation differed after five bands had been formed, and internal bands ceased forming after 30 years. Females to 54 years old and males to 34 years old were examined. Maturity occurred over a wide age range, with estimates of 20 years for females and 9 years for males. The youngest pregnant female was 27 years old. The Francis reparameterized von Bertalanffy growth model found similar growth for males and females, and the von Bertalanffy equations were L _t = 96.12^{(1 – e(–0.072(t+6.13))}) for females and L_t = 73.22(1 – e^{(–0.141(t+2.99))}) for males.

An improved method for the purification of rat liver-type fatty acid binding protein from Escherichia coli

We have developed expedient and reliable methods to isolate cyclosporin synthetase for in vitro biosynthesis of cyclosporins. We have examined enzyme purification strategies suited to large-scale processing and present a chromatographic sequence that serves as a pilot model for industrial scale preparation of cyclosporin synthetase from cyclosporin producing fungi. A chromatographic sequence consisting of ammonium sulfate precipitation → gel filtration → hydrophobic interaction chromatography → anion exchange chromatography, yielded an electrophoretically homogeneous cyclosporin synthetase preparation (Coomassie G-250 brilliant blue staining). Furthermore, a native polyacrylamide gel electrophoresis system was developed for the isolation of active cyclosporin synthetase enzyme from crude extracts of cyclosporin producing fungi. The environmental factors affecting enzyme stability and the continuity of the in vitro cyclosporin biosynthetic reaction-temperature, pH, and substrate depletion were assessed and manageable conditions have been defined for sustainable cyclosporin biosynthesis with enzyme isolates. Cyclosporin synthetase exhibited an optimal temperature range of 24–29 °C and a pH optimum of 7.6. The native enzyme displayed a pI of 5.7, as determined by isoelectric focusing. The industrial implementation of an in vitro biosynthetic approach could potentially prove useful for the production of important therapeutic cyclosporins which occur as only minor fermentation by-products.

The reliability of muscle biopsies taken from vastus lateralis

The purpose of this study was to examine whether a single biopsy sample of vastus lateralis could provide an accurate estimate of capillary density (CD) which is indicative of the entire muscle, or whether capillary density is distributed unevenly and varies with muscle depth. Whole muscle cross sections of vastus lateralis were excised post mortem (n=11) for analysis of capillary density at three muscle depths, (superficial, mid and deep regions). Muscle thickness varied widely (17–79mm) across subjects. The distribution of CD throughout the depth of the muscle was homogeneous in 8 subjects, but in 3 subjects it was heterogeneous (p<0.05). In 3 of these subjects there was a significant (p<0.05) effect of sample depth on CD. These data indicate that tissue from a single biopsy will not adequately represent the CD of the entire vastus lateralis in some individuals. Single biopsies from unspecified muscle depth, have routinely been used to estimate CD and fibre type in vastus lateralis. The present study indicates that a more reliable method of analysis would be to use the tissue from two needle biopsies taken at the superficial and deep portions of the muscle from a group of at least 10 subjects. Sampling theory analysis supported this conclusion.

A design-based method for estimating glomerular number in the developing kidney

Low glomerular (nephron) endowment has been associated with an increased risk of cardiovascular and renal disease in adulthood. Nephron endowment in humans is determined by 36 wk of gestation, while in rats and mice nephrogenesis ends several days after birth. Specific genes and environmental perturbations have been shown to regulate nephron endowment. Until now, design-based method for estimating nephron number in developing kidneys was unavailable. This was due in part to the difficulty associated with unambiguously identifying developing glomeruli in histological sections. Here, we describe a method that uses lectin histochemistry to identify developing glomeruli and the physical disector/fractionator principle to provide unbiased estimates of total glomerular number (Nglom). We have characterized Nglom throughout development in kidneys from 76 rats and model this development with a 5-parameter logistic equation to predict Nglom from embryonic day 17.25 to adulthood (r2 = 0.98). This approach represents the first design-based method with which to estimate Nglom in the developing kidney.

An analytical approach to predict web-warping and longitudinal strain in flexible roll formed sections of variable width

To reduce weight and improve passenger safety there is an increased need in the automotive industry to use Ultra High Strength Steel (UHSS) for structural and crash components. However, the application of UHSS is restricted by their limited formability and the difficulty of forming them in conventional stamping. An alternative method of manufacturing structural auto body parts from UHSS is the flexible roll forming process, which allows the manufacture of metal sheet with high strength and limited ductility into complex and weight-optimized components. One major problem in the flexible roll forming of UHSS is the web-warping defect, which is the deviation in height of the web area over the length of the profile. It has been shown that web-warping is strongly dependant to the permanent longitudinal strain formed in the flange of the part. Flexible roll forming is a continuous process with many roll stands, which makes numerical analysis extremely time intensive and computationally expensive. An analytical model of web-warping is therefore critical to improve design efficiency during the early process design stage before FEA is applied. This paper establishes for the first time an analytical model for the prediction of web-warping for the flexible roll forming of a section with variable width. The model is based on evaluating longitudinal edge strain in the flange of the part. This information is then used in combination with a simple geometrical model to investigate the relationship between web-warping and longitudinal strain with respect to process parameters.

Detecting invertebrate responses to fire depends on sampling method and taxonomic resolution

New knowledge about the responses of species to fire is needed to plan for biodiversity conservation in the face of changing fire regimes. However, the knowledge that is acquired may be influenced by the sampling method and the taxonomic resolution of a study. To investigate these potential sampling biases, we examined invertebrate responses to time since fire in mallee woodlands of southern Australia. Using a large-scale replicated study system, we sampled over 60000 invertebrates with large pitfall traps, wet pitfall traps and sweep nets, and undertook analyses at morphospecies and order level. Large pitfalls and sweep nets detected several strong fire effects, whereas wet pitfall traps detected few effects. Invertebrate abundance in sweep nets was highest shortly after fire because of grasshopper outbreaks. Several additional morphospecies showed strong preferences for different stages in the post-fire succession. In contrast with morphospecies effects, analyses at order level either failed to detect fire effects or were driven by the most abundant species. For fire research to produce credible results with the potential to guide management, it must use a range of sampling techniques and undertake analyses at (morpho)species level. Homogeneous fire management, such as fire suppression in fragmented landscapes or widespread frequent burning for asset protection, is likely to cause declines in fire-affected invertebrates.