On the control of ecdysone biosynthesis by the central nervous system of blowfly larvae

Ecdysone was found to be the major secreted steroid of ring glands dissected from blowfly larvae and incubated in vitro. Other secretory products such as 3-dehydroecdysone and 20-deoxy-makisterone A could not be detected when the glands were labelled with tritiated cholesterol. Ecdysone synthesis and secretion were found to be tightly coupled. The highest rate of secretion was observed a few hours before pupariation. In vitro, the rate of ecdysone secretion by ring glands was affected significantly by coincubation with the central nervous system (CNS). Modulating effects from the CNS to the gland were mediated both by culture medium and by nerve connections. Distinct parts of the CNS revealed multiple and partially opposite effects on ecdysone secretion, suggesting a more complex control than had been anticipated. Multiple neural control systems appear to be involved. Moreover, the observed effects changed with development during the second half of the third instar, reflecting a significant plasticity of neural control. © 1993 Wiley-Liss, Inc.     

The physical characteristics of the members during the International Biomedical Expedition to the Antarctic

Twelve male medical scientists formed the International Biomedical Expedition to the Antarctic (IBEA). Their physical characteristics and maximum oxygen uptakes (VO2max) were measured in association with three series of thermal tolerance tests in Sydney, twice before and once after going to the Antarctic. In the Antarctic they lived in tents and spent 15 days travelling by motor toboggan. Their body mass (BM) and skinfold thickness (SFT) were measured four times during the 69 days the expedition spent in the field. The characteristics of the group were (ranges): age 26-52 years, height 1680-1889 mm, BM 58.5-103.4 kg, fatness 16-34% BM and VO2max 33-49 ml X kg-1 X min-1. In the Antarctic 9 men lost between 0.7 and 5.5 kg (mean 2.7 kg) of BM with a decrease in SFT, whilst 2 men increased BM by 1.2 and 1.9 kg without change in SFT. One man retired early from the expedition. BM and SFT were regained and physical fitness lost during the return voyage to Australia. Consequently there was no difference in average SFT between the pre- and post-Antarctic laboratory tests, but BM was greater after the Antarctic implying gains in fat free mass. VO2max was lower in the final laboratory tests than in the tests before Antarctica.     

Determination of surface immunoglobulin density on frozen-thawed mononuclear cells analyzed by the fluorescence-activated cell sorter

Recent data have demonstrated that differences in sIg density on B lymphocytes distinguish functionally distinct subpopulations of these cells. Other reports suggest that cyropreservation may change the frequency of sIg-bearing lymphocytes. To determine if cryopreservation alters either the frequency of sIg cells or the distribution of sIg density, PBM from normals and patients with CLL and LCL were analyzed using the FACS. Aliquots of Ficoll-Hypaque-separated PBM were controlled-rate frozen (1 °C/min) in 7.5% Me₂SO in RPMI 1640 and thawed in a 37 °C water bath on the same day. Fresh and frozen-thawed PBM aliquots were labeled with fluorescein conjugates of F(ab′) fragments of affinity chromatography-purified anti-Fab or class-specific anti-μ, anti-δ, anti-γ, or anti-α. Histograms of relative cell fluorescence, reflecting sIg density, were prepared for each aliquot with the FACS. The frequency of sIg-bearing PBM labeled with each reagent was not significantly altered by freezing. Likewise, FACS profiles demonstrated that the distribution of sIg density on normal and CLL PBM was unchanged after freezing. However, the fluorescence peak produced by frozen-thawed unlabeled cells was occasionally slightly broader than that of fresh cells, suggesting increased autofluorescence induced by freezing. These data indicate that frozen cell preparations may be utilized for the study of B-lymphocyte subsets as determined by sIg density.     

7-Azaindole-3-acetic acid derivatives: Potent and selective CRTh2 receptor antagonists

High throughput screening identified a 7-azaindole-3-acetic acid scaffold as a novel CRTh2 receptor antagonist chemotype, which could be optimised to furnish a highly selective compound with good functional potency for inhibition of human eosinophil shape change in whole blood and oral bioavailability in the rat.     

IL-15 maintains T-cell survival via S-nitrosylation-mediated inhibition of caspase-3

Caspase activity is critical for both T-cell survival and death. However, little is known regarding what determines caspase activity in cycling T cells. Interleukin (IL)-2 and IL-15 confer very different susceptibilities to T-cell death. We therefore considered that IL-2 and IL-15 differentially regulate caspase activity to influence T-cell survival. We observed that IL-2-cultured primary murine effector T cells manifested elevated levels of caspase-3 activity compared with IL-15-cultured T cells. T cell receptor (TCR) restimulation further increased caspase activity and induced considerable cell death in IL-2-cultured T cells, but provoked only a minimal increase of caspase activity and cell death in IL-15-cultured T cells. IL-2 sensitization to cell death was caspase-3 mediated. Interestingly, increased active caspase-3 levels with IL-2 were independent of active initiator caspase-8 and caspase-9 that were similar with IL-2 and IL-15. Rather, caspase-3 activity was inhibited by posttranslational S-nitrosylation in IL-15-cultured T cells, but not in the presence of IL-2. This paralleled increased reactive nitrogen and oxygen species with IL-15 and reduced glycolysis. Taken together, these data suggest that the metabolic state conferred by IL-15 inhibits T-cell apoptosis in part by maintaining low levels of active caspase-3 via S-nitrosylation.     

Dna2 Is Involved in CA Strand Resection and Nascent Lagging Strand Completion at Native Yeast Telomeres

Post-replicational telomere end processing involves both extension by telomerase and resection to produce 3′-GT-overhangs that extend beyond the complementary 5′-CA-rich strand. Resection must be carefully controlled to maintain telomere length. At short de novo telomeres generated artificially by HO endonuclease in the G₂ phase, we show that dna2-defective strains are impaired in both telomere elongation and sequential 5′-CA resection. At native telomeres in dna2 mutants, GT-overhangs do clearly elongate during late S phase but are shorter than in wild type, suggesting a role for Dna2 in 5′-CA resection but also indicating significant redundancy with other nucleases. Surprisingly, elimination of Mre11 nuclease or Exo1, which are complementary to Dna2 in resection of internal double strand breaks, does not lead to further shortening of GT-overhangs in dna2 mutants. A second step in end processing involves filling in of the CA-strand to maintain appropriate telomere length. We show that Dna2 is required for normal telomeric CA-strand fill-in. Yeast dna2 mutants, like mutants in DNA ligase 1 (cdc9), accumulate low molecular weight, nascent lagging strand DNA replication intermediates at telomeres. Based on this and other results, we propose that FEN1 is not sufficient and that either Dna2 or Exo1 is required to supplement FEN1 in maturing lagging strands at telomeres. Telomeres may be among the subset of genomic locations where Dna2 helicase/nuclease is essential for the two-nuclease pathway of primer processing on lagging strands.     

Biggest challenges in bioinformatics

The third Heidelberg Unseminars in Bioinformatics (HUB) was held on 18th October 2012, at Heidelberg University, Germany. HUB brought together around 40 bioinformaticians from academia and industry to discuss the ‘Biggest Challenges in Bioinformatics’ in a ‘World Café’ style event.