PubMed Health⌕ Search

Biomedical subjects

T Kakefuda

Publications and source records attributed to T Kakefuda.

50 records · Page 3Linked to original sources

Linear, single-stranded deoxyribonucleic acid isolated from Kilham rat virus.

Kilham rat virus (KRV) was grown in a rat nephroma cell line and was purified by two isopycnic centrifugations in cesium chloride. The virus contains single-stranded deoxyribonucleic acid (DNA) with a molecular weight of approximately 1.6 x 10(6). The DNA was extracted from the virion by both phenol extraction and by 2% sodium dodecyl sulfate at 50 C. KRV DNA, extracted by both procedures, was observed in an electron microscope by using a cytochrome c or diethylaminoethyldextran monolayer. The DNA was also exposed to exonuclease I, an enzyme which hydrolyzes specifically linear, single-stranded DNA. Hydrolysis of 70 to 80% of the DNA was observed. Both the enzymatic and the electron microscope studies support the conclusion that extracted KRV DNA is a single-stranded, linear molecule. The length of the DNA was measured in the electron microscope and determined to be 1.505 +/- 0.206 mum.

Animals↗

Electron microscopic observations on the ribonucleic acid of murine leukemia virus.

The ribonucleic acid (RNA) of murine leukemia virus (MLV) Rauscher strain was observed by the aid of electron microscopy with the use of the protein monolayer technique. RNA was observed directly after release from virus particles or after isolation by sedimentation in sucrose density gradients. Molecules were found in an extended linear form. Many of the RNA filaments released by detergent treatment contained curled regions, suggesting the linear filaments were originally coiled within the virus particle. The relationship of the curled areas to the containment of the RNA within the virus particle is discussed, and a mechanism for the inclusion of RNA in the budding virion is proposed. Treatment of the extended MLV-RNA with dimethyl sulfoxide resulted in the collapse of the molecule forming a tangled complex. Treatment with urea or heating at 50 C in 3 mm NaCl also produced this effect. Also under the conditions in which MLV-RNA was linear, RNA from Rous sarcoma virus also was linear, but Newcastle disease virus RNA and ribosomal RNA of rat liver had collapsed structures. The results indicated that the RNA of MLV, and perhaps other RNA-containing tumor viruses, has a specific unique conformation dependent upon hydrogen bonds.

Avian Sarcoma Viruses↗

Ultrastructure of the membrane system in Lactobacillus plantarum.

Electron microscopic study of Lactobacillus plantarum revealed mesosomes in different stages of maturation and structural relation with other cell organelles. Small, immature mesosomes were bounded by a prominent electron-dense layer with another extremely faint layer on the outside. This corresponds to the appearance of the cytoplasmic membrane. Large mature mesosomes were surrounded by a triple-layered unit membrane having electron-opaque layers of approximately equal density, suggesting that the composition of the boundary membrane alters during development of this structure. Three-dimensional observations derived from serial sections indicated that mesosomes always maintain a connection between the cytoplasmic membrane and the comparable layers of their boundary. The cytoplasmic membrane also consisted of a triple-layered unit membrane, the innermost layer of which was less electron-opaque and was usually hidden by the relatively dense background of the cytoplasm. The innermost layer of the cytoplasmic membrane was most clearly seen in plasmolyzed cells. Only mature mesosomes made distinct contacts with, or were partially immersed in, the nucleoplasm. The boundary of such mesosomes frequently seemed to be discontinuous, suggesting that the mesosome interior was in direct contact with the nucleoplasm. Mesosomes involved in cross-wall formation at a division plane increased in size and passed through a sequence of positions which led ultimately to an association with the nucleoplasms of the daughter cells. The inner surface of the cell wall was lined by a thin, electron-dense layer whose composition and function are unknown. Under the cultural conditions used, this organism regularly contained a polyphosphate granule.

Cell Membrane↗

Examination of 7-ethoxycoumarin deethylation and ammonia removal activities in 31 hepatocyte cell lines.

Aiming the selection of possible candidates for the reactor cells in a hybrid artificial liver, we investigated the activities of 7-ethoxycoumarin deethylation and ammonia removal in 31 hepatocytes cell lines established from the human, rat, mouse, and chicken, compared with freshly isolated rat hepatocytes. Rat liver homogenate and hepatocytes showed 7-ethoxycoumarin deethylation activity of 0.925 and 1.840 nmol/mg protein/min, respectively. Two human hepatoblastoma cell lines, Hep G2 and HuH 6, showed the activity of 0.190 and 0.027 nmol/mg protein/min, respectively. While the rat hepatocytes efficiently removed added ammonia form the culture medium, no ammonia removal activity was detectable in any cell line with the assay conditions in this study. Ammonia removal activity, possibly catalyzed by the urea cycle, is likely to be harder to restore than the drug metabolizing system in the course of cell immortalization.

Ammonia↗