Theoretical estimates of cathodoluminescence of giemsa stained chromosomes

The cathodoluminescence (CL) of Giemsa and the recorded cathodoluminescent images of Giemsa-stained chromosomes and chromatin in a scanning electron microscope have been reported. A theoretical estimate of the CL-yield of Giemsa-stained chromosomes is also given. This estimate supports the observed intense CL images. The dye signal has been estimated by taking into consideration the collection and detection efficiencies of a transmission light pipe-photomultiplier system, used in the CL experiments. The calculations have been extended to two other useful dye compounds, viz., fluorescein and acridine orange. Some possible developments of CL in a SEM with these compounds have been suggested.     

Kinetics and mechanism of oxidation of some aldoses by vanadium(v) in perchloric acid medium

The kinetics of oxidation of some aldoses by vanadium(V) in perchloric acid media have been investigated. Each reaction is first order with respect to both [Vanadium(V)] and [Aldose]. The reactions are catalysed by acid. The addition of sodium perchlorate accelerates the rate of reaction. Kinetic evidence for the formation of an intermediate compound between vanadium(V) and aldoses is insignificant, and a mechanism is suggested in which vanadium(V) reacts with the aldoses by a fast step to form a transition state, followed by the decomposition of the latter to give the products of reaction in a slow step. The formation of free-radical intermediates has been demonstrated, and one-electron reduction of vanadium(V) by aldoses seems to be the most plausible mechanism. The oxidation rates follow the order: xyloses arabinose galactose mannose. The activation parameters are reported.     

Release of low density lipoprotein from its cell surface receptor by sulfated glycosaminoglycans.

The sulfated glycosaminoglycan, heparin, was found to release 125I-labeled low density lipoprotein (125I-LDL) from its receptor site on the surface of normal human fibroblasts. Measurement of the amount of 125I-LDL released by heparin permitted the resolution of the total cellular uptake of 125I-LDL at 37 degrees C into two components: first, an initial rapid, high affinity binding of the lipoprotein to the surface receptor, from which the 125I-LDL could be released by heparin, and second, a slower process attributable to an endocytosis of the receptor-bound lipoprotein, which rendered it resistant to heparin release. At 4 degrees C the amount of heparin-releasable 125I-LDL was similar to that at 37 degrees C, but interiorization of the lipoprotein did not occur at the lower temperature. The physiologic importance of the cell surface LDL receptor was emphasized by the finding that mutant fibroblasts from a subject with homozygous Familial Hypercholesterolemia, which lack the ability to take up 125I-LDL at 37 degrees C, did not show cell surface binding of 125I-LDL, as measured by heparin release, at either 4 degrees C or 37 degrees C. Although heparin released 125I-LDL from its binding site, it did not release 3H-concanavalin A from its surface receptor, and conversely, alpha-methyl-D-mannopyranoside, which released 3H-concanavalin A, did not release surface-bound 125I-LDL. When added to the culture medium simultaneously with LDL, heparin prevented the binding of LDL to its receptor and hence prevented the LDL-mediated suppression of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity. The uptake of LDL by fibroblasts is proposed as a model of receptor-mediated adsorptive endocytosis of macromolecules in human cells.     

Preliminary x-ray investigation of an orthorhombic crystal form of human plasma albumin.

An orthorhombic form of single crystals of human plasma albumin, suitable for x-ray diffraction studies, has been grown with ammonium sulfate from protein solutions purified from fresh frozen single donor plasma as well as from a commercial sample of plasma albumin. The space group is P2(1)2(1)2 with 12 molecules in the unit cell. The cell dimensions are: a = 133.3 +/- 1.2 A, b = 274.8 +/- 3.3 A,, and c = 58.02 +/- 0.02 A.     

Studies on the binding of 1-anilino-8-naphthalene sulfonate to very low density and high density himan serum lipoproteins.

Very low density and high density lipoproteins have been isolated from human plasma and their interaction with 1-anilin0-8-naphthalene sulfonate has been studied under different conditions of pH and added salt. Intrinsic fluorescence of bound 1-anilino-8-naphthalene sulfonate was higher for high density lipoproteins then for very low density lipoproteins, but was unaffected by salt in both systems. Binding of 1-anilino-8-naphthalene sulfonate by both these lipoproteins was saturable and was higher in the presence of added NaCl or CaCl2, Ca2+ having a greater effect than Na+ in enhancing fluorescence. The binding data were analyzed by Scatchard plots; the number of binding sites and the affinity of 1-anilino-8-naphthalene sulfonate for the site increased with increasing salt concentration. Fluorescence pH curves were similar to those published for phospholipids. From these and previous observations it is suggested that the phospholipids probably represent the major binding sites for 1-anilino-8-naphthalene sulfonate.     

PURIFICATION AND PROPERTIES OF BRAIN N-ACETYL-β-HEXOSAMINIDASE A

—N-Acetyl-β-hexosaminidase A was purified to homogeneity from human and monkey brains by the conventional procedures followed by concanavalin A–Sepharose affinity chromatography. The optimal activity was observed at pH 4·5 for both enzyme preparations with both the aglycones N-acetylglucosamine and N-acetylgalactosamine derivatives. The K_m values for hexosaminidase A from monkey brain were 0·26 mm and 0·04 mm respectively for N-acetylglucosamine and N-acetylgalactosamine. K_m values obtained for glucosamine and galactosamine derivatives for the human brain hexosaminidase A were of the same order. The glycoprotein nature of the enzymes was established by the affinity towards concanavalin A as well as by the presence of sialic acid, galactose, glucose, mannose and hexosamines in the enzyme molecule from monkey brain.