High resolution two-dimensional gel electrophoresis of the proteins and glycoproteins of human blood platelets and platelet membranes

The proteins and glycoproteins of human blood platelets and platelet membranes in both the reduced and the unreduced states have been analysed by isoelectric focusing and sodium dodecyl sulphate-discontinuous polyacrylamide gel electrophoresis in a two-dimensional technique. Gels which had been stained with periodic acid-Schiff's reagent could be counter-stained with Coomassie Brilliant Blue, simplifying the recognition of components which stain with both reagents. The major glycoproteins and some of the proteins have been identified and the characteristics of the membrane and of the whole platelet components established in this system.     

A 13c-n.m.r. study of 1,6:2,3- and 1,6:3,4-dianhydro-β-D-hexo-pyranoses and their O-acetyl and deoxy derivatives

¹³C-N.m.r. spectra of all possible 1,6:2,3- and 1,6:3,4-dianhydro-β-D-hexo-pyranoses and their O-acetyl and deoxy derivatives are presented. Relations between chemical shifts of certain carbon atoms and the structure of the dianhydrides are outlined, and their application in structural analysis is discussed. Inversion of configuration of the oxirane ring from the endo to the exo position is associated with typical upfield-shifts for oxirane-ring carbon atoms C-2 or C-4, respectively. Possible inter-relationships between ¹³C-chemical shifts and steric and polar interactions in the dianhydro derivatives are discussed.     

Uptake of immune RNA by normal mouse spleen cells

Summary Normal mouse spleen cells take up in vitro radioactively labeled immune RNA. RNA taken up is present in nuclei, polysomes, membranes and cytoplasm. About 20–40% of immune RNA is nonspecifically associated with cell surface. 45% of RNA taken up is degraded and reutilized inside the cells within 2 hours.This work was supported by the Polish Academy of Sciences within the project 09.7.4.1.1.     

Selective lesions of acinar pancreatic cells in rats poisoned with abrin

Rats poisoned with abrin (2.5 micrograms/100 g body weight) died within 36 h with severe necrosis of acinar pancreatic cells. Incorporation in vivo of labelled amino acids into pancreatic protein was greatly impaired 6 h after poisoning. Microsomes isolated from the pancreas of poisoned rats at 6 h had a reduced capacity for protein synthesis in vitro. Incorporation in vivo of orotic acid into pancreatic RNA was decreased 12 h after poisoning.     

Mortality in Grey seal pups: incidence and causes

Pup production and mortality of beach breeding Grey seals was studied at two contrasting types of rookery, the cliff-bound island of Ramsey, Dyfed, and the low, grassy island of Auskerry, Orkney. Thirty-five per cent of pups died on Ramsey compared with 14 % on Auskerry. Almost half of the Ramsey mortality was accounted for by animals being lost from the beaches so that they were not available for analysis. The main proximate causes of death revealed by post-mortem examination were starvation and infections. The other conditions which accounted for a small proportion of deaths were drowning, trauma, non-viability, stillbirths, atelectasis, dystokia and heart abnormality. Certain pathogens associated with disease were common to both sites, while others were specific to Ramsey or Auskerry.
The most important ultimate cause of pup deaths appeared to be failure of the mother/pup bond. The differences in levels of mortality between the sites is thought to be due to differences in the topography of the beaches. Pup survival on the narrow cliff-bound beaches, or in caves, on Ramsey was reduced either directly through pups being washed off, or indirectly by overcrowding of animals at high tide.     

Solubilization of microsomal phosphoethanolaminetransferase by octyl glucoside

Phosphoethanolaminetransferase of high specific activity was solubilized from rat liver microsomes with the non-ionic detergent octyl glucoside. The solubilization method is fast and simple, allowing for processing of large amounts of material. The solubilized enzyme is stable. It contains virtually no phosphocholinetransferase activity. A preliminary characterization of the enzyme, with both diacyl- and alkylacyl-glycerol as substrate, is given. For the reaction, the lipid substrates were incorporated into artificial phospholipid bilayers (liposomes).     

Investigations on the turnover of adrenocortical mitochondria

Summary The effects of chronic administration of ACTH (up to 36 consecutive days) on the mitochondria of the zona reticularis of the rat adrenal cortex were investigated by stereologic techniques. It was found that ACTH induces two phases of hypertrophy of mitochondria alternating with two proliferative stages, which are associated with a significant decrease in the average volume of the organelles. It is suggested that, as in the zona fasciculata, ACTH controls the processes of growth and division of mitochondria in the zona reticularis. The mechanism underlying this action of ACTH as well as the differences between the responses to ACTH of the mitochondrial population of the two adrenal zones are discussed in the light of evidence indicating that mitochondria contain a complete genetic apparatus largely independent of nuclear control.The authors wish to thank Miss A. Coi and Mr. G. Gottardo for their excellent technical assistance. This work was partly supported by a contract with the CNR (C.T. 73.00663.04)     

Linkage between the B-cell specific receptor for monkey red blood cells and HL-A antigens in man-mouse hybrids

Immunogenetics - The established human lymphoid cell lines 6410 and WI-L2 exhibit the recently discovered receptor for monkey red blood cells (MRBC). This receptor is specific for B cells. These...     

A toolbox for class I HDACs reveals isoform specific roles in gene regulation and protein acetylation

The class I histone deacetylases are essential regulators of cell fate decisions in health and disease. While pan- and class-specific HDAC inhibitors are available, these drugs do not allow a comprehensive understanding of individual HDAC function, or the therapeutic potential of isoform-specific targeting. To systematically compare the impact of individual catalytic functions of HDAC1, HDAC2 and HDAC3, we generated human HAP1 cell lines expressing catalytically inactive HDAC enzymes. Using this genetic toolbox we compare the effect of individual HDAC inhibition with the effects of class I specific inhibitors on cell viability, protein acetylation and gene expression. Individual inactivation of HDAC1 or HDAC2 has only mild effects on cell viability, while HDAC3 inactivation or loss results in DNA damage and apoptosis. Inactivation of HDAC1/HDAC2 led to increased acetylation of components of the COREST co-repressor complex, reduced deacetylase activity associated with this complex and derepression of neuronal genes. HDAC3 controls the acetylation of nuclear hormone receptor associated proteins and the expression of nuclear hormone receptor regulated genes. Acetylation of specific histone acetyltransferases and HDACs is sensitive to inactivation of HDAC1/HDAC2. Over a wide range of assays, we determined that in particular HDAC1 or HDAC2 catalytic inactivation mimics class I specific HDAC inhibitors. Importantly, we further demonstrate that catalytic inactivation of HDAC1 or HDAC2 sensitizes cells to specific cancer drugs. In summary, our systematic study revealed isoform-specific roles of HDAC1/2/3 catalytic functions. We suggest that targeted genetic inactivation of particular isoforms effectively mimics pharmacological HDAC inhibition allowing the identification of relevant HDACs as targets for therapeutic intervention.