Study of human erythrocyte membrane proteins by 2-dimensional electrophoresis

Fractionation of human erythrocyte membrane proteins was performed using a modification of two-dimensional gel electrophoresis described by P. O'Farrel with isoelectric point plotted against molecular mass. All major erythrocyte proteins, including high molecular weight proteins, such as spectrin and band 3 protein, identified by one-dimensional sodium dodecyl sulfate gel electrophoresis, were visualized by silver staining of two-dimensional gels. All in all about 50 polypeptides were distinguished on two-dimensional electrophoretic patterns. Preliminary protein map was developed.     

Reversing the detrimental effect of adriamycin on skin grafts in rats

We evaluated in a rat model the effects of a homologous fibrin glue in reversing the effects of Adriamycin on adherence and take of skin grafts. A total of 40 male Fisher rats were used in the study. During the first phase of the experiment, the animals were assigned to either group I (N = 10) receiving normal saline or group II (N = 10) receiving 6 mg/kg Adriamycin by tail vein injection 24 hours before surgery. Skin grafts with and without fibrin glue were placed over wounds in the backs of the animals and adherence was measured at 24 and 48 hours. In the second phase (N = 20), the experiment was repeated, this time evaluating the total area of skin graft take at 7 days. Fibrin glue was found to increase adherence and take of skin grafts in all Adriamycin-treated animals.     

Chemotactic assay for biological effects of silver nanoparticles

A method is proposed for assessing the biocidal efficacy of water-dispersed nanoparticles of silver. It is based on negative chemotaxis of the plasmodia of the slime mold Physarum polycephalum. Biocidal and repellent effects were compared for silver nanoparticles, Ag⁺ ions, and AOT in solution and in the agar gel. In such characteristics as increasing the period of auto-oscillations of contractile activity, decreasing the area of spreading on substrate, and substrate preference in spatial tests, silver nanoparticles proved to be substantially more effective than Ag⁺ and AOT. The lethal concentrations of the nanoparticles were close to those found earlier for bacteria and viruses. The chemotactic tests allow quantitative assessment of the biological reaction and monitoring its dynamics; in resolution, they are superior to the tests based on the lethal action of biocidal agents.     

RNA and DNA in developing retinae: comparison of a normal with the degenerating retinae of C3H mice

Abstract— The Nucleic acids were measured in developing retinae of normal (DBA) mice and those afflicted with an inherited degenerative disease (C3H). The content of RNA in normal and C3H retinae increased to a maximum at 5 days of postnatal age. Thereafter, that of C3H retinae declined to a value lower than the normal. The content of DNA in normal and C3H retinae was maximal at 10 and 5 days of postnatal age, respectively. By 20 days, it declined in both retinae to nearly adult values. The DNA/RNA ratio of normal adult retinae was about 3, while that of C3H adult retinae was nearly 1–5. It is proposed that the photoreceptor cells possess a smaller cytoplasmic volume and a larger DNA/RNA ratio than the cells of the inner retina. The loss of DNA in developing normal and C3H retinae appears to result from cellular death. It was calculated that approximately 1 million cells in normal and 6 million cells in C3H retinae disappear during development. Cellular death in C3H retinae may not be restricted to the photoreceptor population.     

Intestinal Coccidiosis in a Patient with Alpha-chain Disease

During an ultrastructural study of small-intestinal mucosa from a patient suffering from alpha-chain disease organisms were identified within the epithelial cytoplasm which showed the fine structural features of the coccidian group. Though coccidiosis is well recognized as causing a diarrhoeal and often lethal illness in animals it has been neglected as a cause of disease in man. Thus this finding may be significant and warrants further investigation into its possible role in the pathogenesis of alpha-chain disease.     

An analysis of the inactivation of the frog sperm nucleus by toluidine blue

Toluidine blue, applied to frog sperm under appropriate conditions, inactivates specifically the sperm nucleus, leaving the extranuclear parts of the cell undamaged. Thus, the dye-treated spermatozoa stimulate eggs to cleave normally, but contribute no chromosomes to the resulting embryos, which develop as typical gynogenetic haploids. The concentration of dye required to produce this inactivation varies with pH. Measurements made over the entire pH range which can be tolerated by sperm cells showed that in the lower part of the range (5 to 7) the effective dye concentration was about 5 x 10(-6)M; in the intermediate range (7 to 8.5) it was 1 x (-6) to 1 x (-7)M; and for the higher pH values (8.5 to 10.0) it was about 5 x (-8)M. Using sperm suspensions containing 1500 cells per c. mm. these concentrations of dye produced specific inactivation of the sperm nuclei within 7 to 60 minutes at 18 degrees C. Tests of the reversibility of the inactivation were made by transferring the sperm from the dye to a dilute Ringer's solution after a known degree of inactivation had been produced. Following removal of the dye the sperm cells were tested on eggs over a period of 2 hours. During this time there was no indication of a reversal of the inactivation. Microscopic observations of sperm treated with (-5)M or 5 X (-5)M dye show that the dye is taken up by the sperm nucleus, which is faintly but definitely stained. The dye appears to be uniformly distributed in the nucleus, while extranuclear structures remain unstained. Measurements of the amount of dye bound per sperm nucleus indicate that the minimal quantity required for complete inactivation is about 6.7 x (-18) mole, while the maximal amount which can be bound without injury to extranuclear structures is about 1.5 x (-16) mole. The value obtained for the minimal requirement (6.7 X (16) mole = 4 X (6) molecules) suggests that there are roughly 4 million binding sites in the nucleus which, when blocked by dye molecules, somehow prevent the sperm chromosomes from participating in the development of the egg.