Deoxyribonuclease I from rat urine: affinity purification, characterization, and immunochemical studies

Deoxyribonuclease I (DNase I) from rat urine was purified about 3,000-fold to apparent homogeneity with a 14% yield by affinity chromatography utilizing polyguanylic acid-agarose and DNA-cellulose. The purified enzyme preparation was found to contain no other detectable nucleases. Isoelectric focusing electrophoresis revealed that all six isoelectric forms of the enzyme had been purified, and the resulting bands all contained DNase I activity. Quantitative amino acid analysis and N-terminal amino acid sequencing were performed on the purified DNase I. The N-terminal sequence up to the 15th residue of the enzyme was identical to that of rat parotid DNase I. The enzyme was found to be a glycoprotein, containing 1 fucose, 10 galactose, 17 mannose, 12 glucosamine, and at least 3 sialic acid residues per molecule. The isoelectric multiplicity of the enzyme was partly due to differences in the sialic acid content of the isoforms. Gel filtration on Superose 12 and electrophoresis on sodium dodecyl sulfate polyacrylamide gels indicated an approximate molecular mass for DNase I of 32 kDa. The enzyme had an optimum pH of 6.5 and required divalent cations such as Ca2+ for its activity. Its activity was inhibited by 1 mM EDTA and EGTA, but not G-actin. An antibody against the purified enzyme was found to be monospecific against rat urine and the pure antigen, and completely blocked the activity of the purified enzyme.     

Capillary gas-liquid chromatographic separation of oxo and oxo-hydroxy bile acid isomers of theallo and normal series

Capillary gas-liquid chromatographic separation was studied for 60 oxo and oxo-hydroxy bile acids of theallo (5α-H, A/B-trans) and normal (5β-H, A/B-cis) series, which differ from one another in the number, position, and configuration of oxo and hydroxy groups at positions C−3, C−4, C−6, C−7, and/or C−12. Good separation of the isomers was achieved by the use of an aluminum-clad fused-silica capillary column coated with bonded and cross-linked methyl polysiloxane.     

Mechanical and biological properties of bioactive bone cement containing silica glass powder

Silica glass powder (SG-P) made by a fusing-quenching method was added as a second filler to a bioactive bone cement consisting of MgO-CaO-SiO2-P2O5-CaF2 apatite and wollastonite containing glass-ceramic powder (AW-P) and bisphenol-a-glycidyl methacrylate (Bis-GMA)-based resin, to achieve a higher mechanical strength and better handling properties in use. Five types of cement were used, containing different weight ratios of AW-P/SG-P (Group 1 = 100/0; Group 2 = 75/25; Group 3 = 50/50; Group 4 = 25/75; and Group 5 = 0/100) as filler, to evaluate the effect of SG-P content on the biological, mechanical, and handling properties. The total proportion of filler added to the cements was 85% w/w. The compressive, bending, and tensile strengths and fracture toughness of the cements increased with SG-P content. The viscosity of cements also increased with SG-P content, and every cement could be handled manually. The cements were evaluated in vivo by packing the intramedullary canals of rat tibiae. An affinity index was calculated for each cement; this was the length of bone directly apposed to cement expressed as a percentage of the total length of the cement surface. Histological examination of implanted tibiae for up to 26 weeks showed that the affinity indices decreased with SG-P content and that those of all the cement groups increased with time. At 26 weeks, Groups 1 and 2 had almost identical affnity indices (79% and 75%; no significant difference) but those of the other groups remained at <50%. Group 2 had better mechanical and handling properties than Group 1, and an SG-P content in the filler of no more than 25% w/w did not interfere strongly with the bioactivity of the cement.     

Essential role of mitochondrially encoded large rRNA for germ-line formation in Drosophila embryos

In Drosophila, pole cells, the progenitors of the germ line, are induced by the factors localized in the posterior pole region of oocytes and cleavage embryos, or germ plasm. Polar granules in germ plasm are electron-dense structures and have been proposed to contain factors essential for pole cell formation. Mitochondrially encoded large ribosomal RNA (mtlrRNA) has been identified as a component of polar granules. We previously have shown that mtlrRNA is able to rescue embryos that fail to form pole cells as a result of UV irradiation. However, there is a possibility that the function of mtlrRNA is limited to UV-irradiated embryos, and the question of whether mtlrRNA is required for the normal pathway leading to pole cell formation remains unanswered. In this study, we report that the reduction of mtlrRNA in germ plasm by injecting anti-mtlrRNA ribozymes into cleavage embryos leads to their inability to form pole cells. Other components of germ plasm, namely oskar mRNA, germ cell-less mRNA, and Vasa and Tudor proteins appear to be unaffected in these ribozyme-injected embryos. These results support an essential role for mtlrRNA in pole cell formation. We propose that mitochondrially encoded molecules participate in a key event in early cell-type specification.     

The 24-hour rhythms in plasma growth hormone, prolactin and thyroid stimulating hormone: effect of sleep deprivation

Twenty-four hour secretory rhythms of growth hormone (GH), prolactin (PRL) and thyroid stimulating hormone (TSH) were investigated in 9 normal adult men by means of serial blood sampling at 30 min intervals. The profiles of pituitary hormones were compared in 6 subjects between in normal nocturnal sleep condition and in delayed sleep condition. Plasma GH was measured with use of highly sensitive enzyme immunoassay (EIA) recently developed. Plasma TSH was also evaluated by highly sensitive time-resolved fluorometric immunoassay (TR-FIA). Time series analysis of plasma GH and PRL was performed by auto- and cross- correlation and spectral analysis. The detection limit of EIA for GH was 0.3 pg/ml and all plasma GH levels were within the detectable range of this EIA. Cross-correlation and spectral analysis suggested the presence of approximately 2-3 h rhythmicity of plasma GH. Plasma PRL appeared to have some 24-hour rhythmicity besides its sleep-dependent component. Sleep deprivation caused marked elevation of plasma TSH during night time. It is suggested that there appears two mechanisms regulating GH secretion: one has a sleep-independent and ultradian rhythm and another has a sleep-dependent rhythm.     

Measurements of complex antenna factor by the near-field 3-antennamethod

The near-field 3-antenna method for measuring the complex antenna factor (CAF) is proposed in order to reduce the effect of the measurement site and background noise. The transmission coefficients in the far-field region are calculated from those measured in the near-field and theoretically obtained near-field correction factors (NCF). The CAF of monopole antennas are measured using the proposed method in a frequency range up to 6 GHz. From the measurement results, the dependency of CAF values on the antenna distance is small. These results indicate that our theory including the NCF is applicable for the measurements of the CAF of simple-structure antennas     

Clinical study on hepatocellular carcinoma with extrahepatic malignancies

In a consecutive series of 146 patients with hepatocellular carcinoma (HCC), 10 patients (6.8%) were found to have one or two extrahepatic malignancies (EHM). Of these, eight had double cancers and two, triple cancers. The associated malignancies included eight cases of gastric cancer and four cases of colon cancer. Among the 12 lesions, eight were in the early stage. All the 10 patients were hepatitis B surface antigen negative. The incidence of coexisting liver cirrhosis and the retention rate of indocyanin green in 15 minutes among HCCs with EHM were significantly lower than those among HCC alone. These results suggest that the etiology of HCC with EHM is different from the etiology of HCC alone in Japan.     

Frictional properties of structural steel JN2 at cryogenic temperatures in a vacuum

To assess the frictional properties at the wedge of a toroidal field coil of a tokamak fusion reactor, friction tests were conducted using structural steel, JN2, against various materials in a vacuum at low temperatures under a normal load of 10 N and a peak-to-peak sliding amplitude of 100 μm. The temperature was mainly ≈ 5 K and the ambient pressure ≈ 10⁻³ Pa. The relationship between the frictional characteristics and the number of cycles depends on both the material combination and temperature. Two typical patterns of behaviour were observed at 5 K depending on the combination; fairly constant friction and very high and fluctuating friction. The high friction was caused by severe adhesion between surfaces. Temperature dependence of friction was also observed for JN2-copper.