The influence of pH on the EPR and redox properties of cytochrome c oxidase in detergent solution and in phospholipid vesicles

The EPR signals of oxidized and partially reduced cytochrome oxidase have been studied at pH 6.4, 7.4, and 8.4. Isolated cytochrome oxidase in both non-ionic detergent solution and in phospholipid vesicles has been used in reductive titrations with ferrocytochrome c.The g values of the low- and high-field parts of the low-spin heme signal in oxidized cytochrome oxidase are shown to be pH dependent. In reductive titrations, low-spin heme signals at g 2.6 as well as rhombic and nearly axial high-spin heme signals are found at pH 8.4, while the only heme signals appearing at pH 6.4 are two nearly axial g 6 signals. This pH dependence is shifted in the vesicles.The g 2.6 signals formed in titrations with ferrocytochrome c at pH 8.4 correspond maximally to 0.25–0.35 heme per functional unit (aa₃) of cytochrome oxidase in detergent solution and to 0.22 heme in vesicle oxidase. The total amount of high-spin heme signals at g 6 found in partially reduced enzyme is 0.45–0.6 at pH 6.4 and 0.1–0.2 at pH 8.4. In titrations of cytochrome oxidase in detergent solution the g 1.45 and g 2 signals disappear with fewer equivalents of ferrocytochrome c added at pH 8.4 compared to pH 6.4.The results indicate that the environment of the hemes varies with the pH. One change is interpreted as cytochrome a₃ being converted from a high-spin to a low-spin form when the pH is increased. Possibly this transition is related to a change of a liganded H₂O to OH^? with a concomitant decrease of the redox potential. Oxidase in phosphatidylcholine vesicles is found to behave as if it experiences a pH, one unit lower than that of the medium.     

Growth of Streptomyces bambergiensis and glucose isomerase biosynthesis

Summary Growth and glucose isomerase biosynthesis in Streptomyces bambergiensis ATCC 13879 have been studied under different conditions. Some data concerning correlation between cultivation conditions and elemental analysis of the cells are also presented.     

Molecular distribution of zinc in biliary and pancreatic secretions

Rat bile and pancreatic fluid were examined for the presence of low molecular weight zinc complexes. Fluids were collected separately by cannulation, and zinc distribution in collected samples was analyzed by gel filtration on Sephadex G-50. Most of the zinc in bile was associated with low molecular weight zinc complexes; only a small amount of zinc was present in the high molecular weight fraction. In contrast, pancreatic secretions did not contain low molecular weight zinc complexes, but there were considerable amounts of zinc bound to high molecular weight compounds. The addition of zinc to bile resulted in an increased amount of zinc in the low molecular weight fraction, while the addition of zinc to pancreatic fluid resulted primarily in an increase in zinc bound to the high molecular weight components. Like pancreatic fluid, homogenates of pancreatic tissue had no low molecular weight zinc complex. In rats whose bile and pancreatic fluid were removed and not returned into the intestine, the amount of zinc bound to low molecular weight complexes in intestinal homogenates was reduced. This alteration of the molecular distribution of zinc in intestinal homogenates by removal of bile and pancreatic fluid suggests the potential importance of low molecular weight zinc complexes for zinc homeostasis.     

Superoxide dismutase isoenzymes in bovine and human milk

The presence of superoxide dismutase in bovine and human milk was investigated by ultrafiltration, gel filtration, and isoelectric focusing. Conclusive evidence for the presence of this enzyme in both milks is presented. The molecular weight of the enzyme was estimated by gel filtration on Sephadex G-100 to be 30,000, which is consistent with reported values for the copper, zinc form of superoxide dismutase. In addition, enzyme activity was inhibited by cyanide, thus eliminating the possibility that the enzyme was present in the manganese form. Several isoenzymes were detected by isoelectric focusing in polyacrylamide gel, and the isoenzyme pattern in bovine milk was the same as that found for bovine plasma, suggesting that milk superoxide dismutase originates from plasma. It may be that the presence of copper, zinc superoxide dismutase in milk is important for the maintenance of its oxidative stability.     

Differential behavior of cytotoxic effector cells against HLA antigens in strong genetic linkage disequilibrium

Five sets of cytotoxic effector cells were generated, using haploidentical, first degree relatives in five different families, against the HLA-A3; B7 serological determinants combined with different DR antigens. When tested against a panel of cells bearing combinations of the HLA-A, -B and -DR antigens it was shown that the HLA-B7 antigen was as strong a CML target determinant alone as it was in the presence of HLA-A3. The strength of the HLA-A3 antigen as target determinant varied. With effector cells primed to the HLA-A3; B7; DR2 haplotype, the A3 antigen alone behaved as a weak target determinant. When the same target cells were tested with the effector cells generated against HLA-A3; B7 without DR2, the A3 antigen behaved as a strong target determinant. A number of target cells lacking the serologically detectable HLA determinants present on the sensitizing HLA haplotype were identified as being killed by specific effector cells. These data suggest either a number of new CML target determinants controlled by different loci or the presence of a single, new locus with multiple alleles controlling CML targets.     

Influence of ashing techniques on the analysis of trace elements in biological samples

Dry ashing and wet ashing are two commonly used methods for the preparation of biological materials for trace element analysis by atomic absorption spectrophotometry. In this paper, National Bureau of Standards (NBS) bovine liver was dry ashed at 450°C for 24 h in silica glass (Vycor) or procelain crucibles; the resulting ash was dissolved in either concentrated nitric or hydrochloric acid. Dry ashing efficiency was evaluated by comparing iron, copper, zinc, and manganese concentrations of the samples with the values certified by NBS. Highest recoveries were obtained by dry ashing in silica glass (Vycor) crucibles. Dissolving the resultant ash in either hydrochloric or nitric acids did not significantly alter the results. A comparison between dry and wet ashing shows the latter method to be superior for the preparation of biological tissues for analysis of iron, copper, zinc, and manganese.     

Visna Virus Meningoencephalomyelitis in Goats

A progressive paresis was encountered in herds of Swedish goats. The symptoms developed during a period of weeks or months, and were initially often seen as a weakness of the hind limbs before the animals became paralytic. The development and the histopathological lesions of the disease in the GNS and the lungs were similar to those of visna in sheep. In vitro grown choroid plexus cells, prepared from affected goats, showed foci of polykaryocytes. Electron microscopy revealed the presence of particles morphologically similar to those of sheep visna virus (SVV). Goats experimentally infected with the goat visna virus (GVV) developed GNS lesions similar to those of visna in sheep and became seropositive to SVV. The results of complement fixation tests, carried out on sera from 11 goat herds, showed a coincidence between seropositiveness and the occurrence of disease in one and the same herd. Using the ELISA method, an average of 80 % of the goats in 5 herds were found to be seropositive to GVV.     

The primary structure of Escherichia coli K12 2-deoxyribose 5-phosphate aldolase. Nucleotide sequence of the deoC gene and the amino acid sequence of the enzyme

The sequence of the deoC gene of Escherichia coli K12 and the amino acid sequence of the corresponding protein, deoxyriboaldolase, has been established. The protein consists of 259 amino acids with a molecular weight of 27 737. The purified enzyme may exist both as a monomer and as a dimer. On the basis of amino acid composition, molecular weight and catalytic properties, the enzymes from E. coli and Salmonella typhimurium seem to be almost similar. They belong to the class I aldolases, which form Schiff base intermediates. Using data for the S. typhimurium enzyme, the lysine residue involved in the active site in the E. coli enzyme was tentatively identified.     

扑虱灵对稻飞虱及其捕食性天敌种群的影响

扑虱灵对稻飞虱的防治效果,及对捕食性天敌的毒性,进行了田间试验,并与叶蝉散,杀虫双的使用结果作比较．试验结果表明,扑虱灵对稻飞虱的防治效果达93％,与叶蝉散,杀虫双相比较。防治效果提高了2．39～32．85％．对黑肩绿盲蝽和蜘蛛种群具有不同程度的选择性,维护较低的稻飞虱与天敌的比值,有利于天敌作用的发挥,维持对稻飞虱较长的残效期,基本上可以控制稻田后期飞虱的增长．     

Nuclear magnetic tomography in the diagnosis of spinal cord cysts]

Magnetic resonance tomography (MRT) was used in examinations of 172 patients with spinal cord abnormalities. Thirty-eight cysts were detected: 25 in syringomyelia, 5 in intramedullary tumors, 4 posttraumatic and 4 postoperational ones. Based on the MRT patterns, two types of syringomyelia were distinguished, the occlusive and idiopathic ones. The size and site of cyst, as shown by the MRT, did not conform to its neurologic symptoms. A characteristic feature of tumorous cysts was a highly intensive signal originating from their contents. Traumatic cysts were detected against the background of diffuse atrophy of the cord. Computer-aided tomography did not give full-value information in such cases and therefore could not be recommended in cases with suspected intramedullary cysts.