Studies of a synthetic substrate in canine globoid cell leukodystrophy

Gal et al. ((1977) Clin. Chim. Acta 77, 53–59) reported the use of a new synthetic substrate, 2-hexadecanoylamino-4-nitrophenyl-β-D-galactopyranoside for the diagnosis of human globoid cell leukodystrophy. Assay of β-galactosidase in brain homogenates from normal, carrier, and globoid cell leukodystrophy-affected dogs utilizing this new substrate demonstrated overlapping activities. Instead of reflecting specific $\text{D}\text{-galactosyl}\text{-N-}\text{acylsphingosine}$ galactohydrolase (EC 3.2.1.46), the 2-hexadecanoylamino-4-nitrophenyl-β-D-galactopyranoside β-galactosidase activity in canine brain is highly correlated with nonspecific 4-methylumbelliferyl β-galactosidase. Optimization of the 2-hexadecanoylamino-4-nitrophenyl-β-D-galactopyranoside assay system for canine brain and the use of varying concentrations of taurocholate or taurodeoxycholate in the assay mixture did not alter the lack of specificity. These results indicate a significant difference in the nature of the underlying defect in galactosylceramide β-galactosidase in canine globoid cell leukodystrophy compared to human globoid cell leukodystrophy.     

The mechanism of the hyperglycaemic action of synthetic peptides related to the C-terminal sequence of human growth hormone

Synthetic part sequences of human pituitary growth hormone (hGH 176–191 and hGH 177–191) corresponding to residues 176–191 or 177–191 of the hormone have been tested for their effects on glycogen and pyruvate metabolism in the rat, both in vivo and in vitro. When injected, the peptides caused transient increases in blood glucose and lactate, while decreasing the activity ratio of glycogen synthase in muscle, adipose tissue and liver and of pyruvate dehydrogenase in muscle and adipose tissue, but not in liver. These decreases were associated with the conversion of the enzymes from their active to their inactive forms, since the peptides did not affect the total amount of either the synthase or the dehydrogenase. The time course of the effect on the enzymes was similar to that for the effect on blood metabolites, and responses for synthase were produced over the range 0.07–7 nmols hGH 177–191/kg body weight. Phosphorylase activity was not affected by the peptides, nor was the capacity to dispose of injected L-lactate. Experiments with adipocytes and hepatocytes showed that the peptides also affected glycogen synthase and pyruvate dehydrogenase activities in vitro. The peptides had no effect on the overall rate of gluconeogenesis from lactate by hepatocytes. However, at times corresponding to those at which glycogen synthase was inactivated, the peptides caused increased incorporation of lactate into free glucose and decreased incorporation into glycogen. It was concluded that the peptides acted directly on their target tissues, and that the observed hyperlactataemia was the result of the inactivation of pyruvate dehydrogenase. The addition lactate increased the flux through the gluconeogenic pathway, and appeared as glucose because the peptide also inactivated glycogen synthase. Thus, the hyperglycaemia produced by hGH 177–199 and related peptides is explicable in terms of a modified Cori Cycle.     

Biophysical studies of cholesterol effects on chromatin

Changes in chromatin structure regulate gene expression and genome maintenance. Molecules that bind to the nucleosome, the complex of DNA and histone proteins, are key modulators of chromatin structure. Previous work indicated that cholesterol, a ubiquitous cellular lipid, may bind to chromatin in vivo, suggesting a potential function for lipids in modulating chromatin architecture. However, the molecular mechanisms of cholesterol's action on chromatin structure have remained unclear. Here, we explored the biophysical impact of cholesterol on nucleosome and chromatin fibers reconstituted in vitro and characterized in silico the cholesterol binding to the nucleosome. Our findings support that cholesterol assists 10 and 30 nm chromatin formation and induces folding of long chromatin fibers as a result of direct interaction of the cholesterol to six nucleosomal binding sites.     

Biological Aspects of Medorippe lanata (Linnaeus, 1767) (Brachyura: Dorippidae) from the Eastern Ligurian Sea (Western Mediterranean)

The aim of the present study is to investigate the demographic structure and to identify some aspects of the biology of an exploited population of Medorippe lanata (Brachyura: Dorippidae) in the eastern Ligurian Sea, western Mediterranean. 1364 specimens (639 males and 725 females) of M. lanata were collected on a monthly basis from January to December 2001, in a wide area of the eastern Ligurian Sea usually exploited by the Viareggio ‘rapido’ trawl fleet. M. lanata represented an important fraction of the discard, both in weight and in number of individuals. Maximum abundance of this species occurred in late summer-early autumn (up to 3369 ind. km⁻² and 50.6 kg km⁻² in August). The overall females:males sex-ratio was 1.13:1, while the monthly sex-ratio did not differ statistically from 1:1 in all months, except in September and October, when females significantly outnumbered males. The sampled population was composed of two cohorts from November to April. Sizes ranged from 10 to 29 mm carapace length (CL) for females and from 9 to 29 mm CL for males. The von Bertalanffy growth curve, computed for both sexes, gave a higher growth rate in males than in females. Recently moulted males and females were observed throughout the year, except in summer, when the highest number of ovigerous females was present. Females with external eggs were collected from March to November, with peaks in August and September. The monthly evolution of the ovarian maturity stages showed no clear temporal trend. At 21 mm CL, 50% of females were ovigerous or showed macroscopically mature ovaries. According to the dimorphism in chelae size, the presence of adult males (post-puberty stage) was observed all year round, from 18 to 29 mm CL, without evident temporal trends.     

Meeting report: Second international meeting on quadruplex DNA

A two and a half day meeting on G-quadruplexes was held in Louisville, KY, USA (April 18–21, 2009). A specific goal of this conference was to promote discussion on the biology of G-quadruplexes. In practice this was represented in four main ways, namely in biophysics, bio/nanotechnology, therapeutics, and what might be termed “intrinsic biology”. Research into the basic biophysical and structural properties of G-quadruplexes continues to be important for understanding biology, and for optimizing aptamers for therapeutic and bio/technological purposes. The meeting comprised two Keynote lectures, twenty-three invited talks, and forty-two posters covering various aspects of these topics using a wide variety of technologies.     

Reproduction of the rare monocarpic species Saxifraga mutata L.

The aim of the study is to investigate the impact of reproduction and genetic variation on the persistence of populations of the prealpine, monocarpic Saxifraga mutata L. The species grows on erosion slopes or rocks, and its local populations are often small and isolated. Crossing experiments resulted in better seed-set than selfing, but both yielded viable seeds. Agamospermy did not occur. In an early-successional species like S. mutata , successful selfing is important in the colonization of new habitats. Flowers of S. mutata were visited by Syrphidae and unspecialized Hymenoptera. A germination rate of 40% was reached in cultivation after 20 weeks but germination continued until the end of the experiment after 92 weeks. Seeds stored dry for 30 months at room temperature mostly lost their germinability. In natural habitats, seedlings were found almost throughout the year with a peak in spring. Suitable safe sites were small patches of open soil, bare marl on erosion slopes, and rock crevices. AU individuals investigated were diploid with 2n = 26. Allozyme electrophoresis showed a lack of segregation within the populations. Intra- and interpopulation genetic variation was low. These results were in partial disagreement with theoretical expectations in a mixed mating species. It is concluded that demographic rather than genetic processes are the main cause of extinction of populations of S. mutata , at least in the short-term.     

Integrating biotechnology into a non-majors biology curriculum

A general education biology course entitled ‘Biotechnology Transforms Our World’ has been developed to illustrate biological concepts with advances from biotechnology. The contributions of molecular biology to understanding human genetics, evolution, plant and animal (including human) biology and ecology are illustrated with specific case studies. Journal of Industrial Microbiology & Biotechnology (2000) 24, 308–309. Received 02 April 1999/ Accepted in revised form 11 November 1999     

Heritable Epigenetic Changes Alter Transgenerational Waveforms Maintained by Cycling Stores of Information

Our view of heredity can potentially be distorted by the ease of introducing heritable changes in the replicating gene sequences but not in the cycling assembly of regulators around gene sequences. Here, key experiments that have informed the understanding of heredity are reinterpreted to highlight this distortion and the possible variety of heritable changes are considered. Unlike heritable genetic changes, which are always associated with mutations in gene sequence, heritable epigenetic changes can be associated with physical or chemical changes in molecules or only changes in the system. The transmission of cycling stores along the continuous lineage of cells that connects successive generations creates waves of activity and localization of the molecules that together form the cell code for development in each generation. As a result, heritable epigenetic changes can include any that can alter a wave such as changes in form, midline, frequency, amplitude, or phase. Testing this integrated view of all heritable information will require the concerted application of multiple experimental approaches across generations.