Increased membrane-associated phorbol-12,13 dibutyrate (PDBu) receptor function in sickle red cells

A four-fold increase in the binding of 3H-PDBu by red cell membrane ghosts isolated from sickle red cells compared to that from normal controls is presented. Phosphorylation studies with gamma-32P-ATP indicate a similar (two to three-fold) increase in the radiolabelling of the acid-precipitable membrane proteins in sickle red cells. When red cells were loaded with Ca2+ using Ionophore A23187, both normal and sickle red cells enhanced their phosphorylation and sickle red cells to a greater extent than normal red cells. Polyacrylamide slab gel electrophoretic separation of the phosphoproteins and autoradiography also reveal phosphorylation, predominantly of protein bands 3, 4.1 and 4.9 which are known in the red cells as specific substrates for the PDBu receptor, protein kinase C. These results indicate that membrane association of protein kinase C in sickle red cells is increased, possibly as a consequence of the pathological change in their ability to accumulate intracellular calcium.     

Effect of substrate stiffness on early human embryonic stem cell differentiation

There is growing demand for robust DNA assembly strategies to quickly and accurately fabricate genetic circuits for synthetic biology. One application of this technology is reconstitution of multi-gene assemblies. Here, we integrate a new software tool chain with 2ab assembly and show that it is robust enough to generate 528 distinct composite parts with an error-free success rate of 96%. Finally, we discuss our findings in the context of its implications for biosafety and biosecurity.     

Synthesis and desulfurization-decarboxylation of cyclic thionocarbonates of glycerol 1-ethers

A method is described for the synthesis of cyclic thionocarbonates of 1-O-alkyl glycerols in quantitative yield. These derivatives of glycerol ethers can be quantitated by UV absorbance, analyzed by gas-liquid chromatography, or quantitatively converted to allyl alkyl ethers for gas-chromatographic analysis.