Unusual peroxidase activity of polynitroxylated pegylated hemoglobin: Elimination of H2O2 coupled with intramolecular oxidation of nitroxides

Polynitroxylated hemoglobin (Hb(AcTPO)₁₂) has been developed as a hemoglobin-based oxygen carrier. While Hb(AcTPO)₁₂ has been shown to exert beneficial effects in a number of models of oxidative injury, its peroxidase activity has not been characterized thus far. In the blood stream, Hb(AcTPO)₁₂ undergoes reduction by ascorbate to its hydroxylamine form Hb(AcTPOH)₁₂. Here we report that Hb(AcTPOH)₁₂ exhibits peroxidase activity where H₂O₂ is utilized for intramolecular oxidation of its TPOH residues to TPO. This represents an unusual redox-catalytic mechanism whereby reduction of H₂O₂ is achieved at the expense of reducing equivalents of ascorbate converted into those of Hb(AcTPOH)₁₂, a new propensity that cannot be directly associated with ascorbate.     

Metal-dependent dimensionality in coordination polymers of a semi-rigid dicarboxylate ligand with additional amide groups: Syntheses, structures and luminescent properties

Three new coordination polymers, [CdL(H₂O)₃·H₂O]_n (1), [MnL(H₂O)₂]_n (2) and [ZnL]_n (3) (L = 2,2′-[1,4-bis(-benzamido)]diacetate) have been hydrothermally synthesized. Complex 1 and complex 2 are 1-D infinite zigzag chain and 2-D rectangular grid networks, respectively, in which are further stabilized by hydrogen bonds, thus affording two 3-D supramolecular frameworks. Complex 3 exhibits a (4, 4)-connected PtS topology. Moreover, an unusual phenomenon has been observed that the dimension of these coordination polymers gradually increases with the decrease of coordination numbers of metal ions. In addition, complex 1 exhibits a strong blue luminescence in the solid state at room temperature and may be potential candidate for luminescent materials.     

Ultrastructure of long-range transport carriers moving from the trans Golgi network to peripheral endosomes

The delivery of mannose 6-phosphate receptors carrying lysosomal hydrolases from the trans-Golgi network (TGN) to the endosomal system is mediated by selective incorporation of the receptor-hydrolase complexes into vesicular transport carriers (TCs) that are coated with clathrin and the adaptor proteins, GGA and AP-1. Previous electron microscopy (EM) and biochemical studies have shown that these TCs consist of spherical coated vesicles with a diameter of 60-100 nm. The use of fluorescent live cell imaging, however, has revealed that at least some of this transport relies on a subset of apparently larger and highly pleiomorphic carriers that detach from the TGN and translocate toward the peripheral cytoplasm until they meet with distally located endosomes. The ultrastructure of such long-range TCs has remained obscure because of the inability to examine by conventional EM the morphological details of rapidly moving organelles. The recent development of correlative light-EM has now allowed us to obtain ultrastructural 'snapshots' of these TCs immediately after their formation from the TGN in live cells. This approach has revealed that such carriers range from typical 60- to 100-nm clathrin-coated vesicles to larger, convoluted tubular-vesicular structures displaying several coated buds. We propose that this subset of TCs serve as vehicles for long-range distribution of biosynthetic or recycling cargo from the TGN to the peripheral endosomes.     

Effects of G-protein probes on interactions between auxin efflux carriers and phytotropin receptors in zucchini (Cucurbita pepo L.) microsomal membranes

Microsomal vesicles prepared from etiolated hypocotyl tissue of zucchini (Cucurbita pepo L. cv. All Green Bush) exhibited saturable N-1-naphthylphthalamic acid ([³H]NPA) binding, NPA-stimulated association of indol-3yl-acetic acid ([³H]IAA), and saturable binding of guanosine 5-O-[3-thiotriphosphate] (GTP--[³⁵S]). These vesicles were used to test the possibility that NPA receptors might interact with IAA-anion efflux carriers by coupling through a GTP-binding protein (G-protein). Unlabelled GTP--S or guanosine 5-O-[2-thiodiphosphate] (GDP--S) had no effect on saturable NPA binding or on the NPA-stimulated association of IAA with microsomes. NPA did not affect saturable binding of GTP--[³⁵S] to microsomes, either in the presence or absence of saturating concentrations of unlabelled GTP--S or GDP. It is concluded that the occupancy of phytotropin receptors is not transduced to auxin efflux carriers by a GTP-binding protein.     

芦苇叶中Rubisco对干旱生境的响应

从分布于甘肃省河西走廓的沼泽芦苇和沙丘芦苇叶中提纯的Ｒｕｂｉｓｃｏ,大、小、小亚基分子量无明显变化,但沙丘芦苇与沼泽芦苇与沼泽芦苇相比较,Ｒｕｂｉｓｃｏ的氨基酸组成中,亲水性氨基酸相对含量增加,疏水性氨基酸相对含量减少,酶分子的疏水性降低,且酶分子中可被ＰＣＭＢ滴定的ＳＨ基数仅为沼泽芦苇的５０％。这说明,芦苇在长期适应沙丘干旱生境过程中,其Ｒｕｂｉｓｃｏ无论在结构与功能上,均发生有适应性的改变。     

兰州市婴幼儿艰难梭菌带菌状况

在兰州市随机采集的２１６名健康婴幼儿粪标本经分离并对其培养特性、菌落菌体形态特征、生化反应特性以及毒素原性等进行一系列检查,检出了３７人的粪便含有艰难梭菌（１０４─１０８／ｇ）,检出总阳性率为１７．１％,新生儿、婴儿及幼儿各年龄组的检出阳性率分别为１３．５％（１９／４１）、３３．３％（１３／３９）、１３．９％（５／３６）。３７株分离菌中毒素原性阳性者仅有８株（２１．６％）,均分布于婴儿与幼儿两组。新生儿１４１人中２７人的粪标本为胎粪,只有１人（３．７％）含艰难梭菌（２×１０６／ｇ）,但不产毒素。所有婴幼儿中有２２人因各种原因曾用过抗菌药物,但仅有２人的粪便含艰难梭菌,而且均为非产毒株,表明艰难梭菌检出率与药物服用之间似无显著的相关性。     

Effects of Doubled CO2 on Contents of Photosynthetic and on Kinetic Parameters of Fluorescence Induction in Different Genotypes of Soybean

Effects of doubled C02 on photosynthetic characteristics of soybean ( Glycine max L. ) Bragg (wild type) and its different monogene mutation strains--Nts 382 (supemodulation mutant) and Nod 49 (non-nodulation mutant) were studied. The experimental results showed that the contents of chlorophyll and carotenoid of Bragg, Nts 382 and Nod 49 were increased under doubled CO2, respectively, although to different extent. The determination results of fluorescence induction kinetic parameters showed that PS Ⅱ activity, the efficiency of primary conversion of light energy of PS Ⅱ and the efficiency of potential photosynthetic quantum conversion of a leaf from Bragg and its mutants were raised in doubled CO2. Fluorescence photochemical quenching coefficient and the overall photochemical quantum yield of PS Ⅱ were raised and non-photochemical quenching coefficient was reduced with CO2 enrichment; such changes were bigger in Nts 382 than those in Bragg and Ned 49. It might be that atmospheric N2 was more effectively utilized by Nts 382 than by Bragg and Ned 49.     

芦苇耐脱水能力的生理生态学分析

甘肃省河西走廊分布有不同生态环境的芦苇,包括沼泽芦苇、盐化草甸芦苇和沙丘芦苇。盐化草甸芦苇和沙丘芦苇表观出很强的耐脱水能力。 测定与植物耐脱水能力相关的SOD、CAT和POD活力,表现为SOD活力以沙丘芦苇最高,盐化草甸芦苇次之,沼泽芦苇最低,但CAT和POD则以沼泽芦苇高于其它生境芦苇。用聚丙烯酰胺垂直板电泳方法分析这3种酶同工酶谱,结果表明,3种不同生境芦苇的SOD同工酶谱基本相同,均有9条酶带,但沙丘芦苇和盐化草甸芦苇的CAT和POD同工酶带数比沼泽芦苇分别增多3条和2条。这些结果证明,芦苇在自然选择压力下形成的耐脱水能力与基因调控的酶多型性相关。     

Spatial development of the cultivation of a bone marrow stromal cell line in porous carriers

The spatial development of the cultivation of a bone marrow stromal cell line (SR-4987) in porous carriers was investigated in order to construct a three-dimensional hematopoietic culture system. Low-rate continuous agitation, 20 rpm, was an appropriate method to achieve initial adhesion of cells onto a cellulose porous beads (CPB, 100 μm pore diameter) in a spinner bottle, compared with other methods such as centrifugation and intermittent agitation. Cell growth with continuous agitation at 70 rpm after initial cell adhesion was not inferior to that at 20 rpm. A 2- and 10-fold increase in the inoculum cell concentration for CPB and another type of porous cellulose beads (Micro-cube (MC), 500 μm pore diameter) resulted in a 1.2- and 2-fold increase in final cell concentrationm, respectively. Cells attached to the MC beads and a polyester nonwoven dic (Fibra-cell (FC)) could grow and spread well on the carriers and a fibroblast-like shape was observed under scanning electron microscopy while the cells on CPB were globular. The flatness and inner surface area of these carriers may be the reason for the differences in cell morphology. This revised version was published online in July 2006 with corrections to the Cover Date.     

Involvement of mitochondrial inner membrane anion carriers in the uncoupling effect of fatty acids

This paper considers stages of the search (initiated by V. P. Skulachev) for a receptor protein for fatty acids that is involved in their uncoupling effect. Based on these studies, mechanism of the ADP/ATP antiporter involvement in the uncoupling induced by fatty acids was proposed (Skulachev, V. P. (1991) FEBS Lett., 294, 158– 162). New data (suppression by carboxyatractylate of the SDS-induced uncoupling, pH-dependence of the ADP/ATP and the glutamate/aspartate antiporter contributions to the uncoupling, etc.) led to modification of this hypothesis. During discussion of the uncoupling effect of fatty acids caused by opening of the Ca²⁺-dependent pore, special attention is given to the effects of carboxyatractylate added in the presence of ADP. The functioning of the uncoupling protein UCP2 in kidney mitochondria is considered, as well as the diversity observed by us in effects of 200 µM GDP on decrease in under the influence of oleic acid added after H₂O₂ (in the presence of succinate, oligomycin, malonate). A speculative explanation of the findings is as follows: 1) products of lipid and/or fatty acid peroxidation (PPO)modify the ADP/ATP antiporter in such a way that its involvement in the fatty acid-induced uncoupling is suppressed by GDP; 2) GDP increases the PPO concentration in the matrix by suppression of efflux of fatty acid hydroperoxide anions through the UCP (Goglia, F., and Skulachev, V. P. (2003) FASEB, 17, 1585–1591)and/or of efflux of PPO anions with involvement of the GDP-sensitive ADP/ATP antiporter; 3) PPO can potentiate the oleate-induced decrease in due to inhibition of succinate oxidation.Translated from Biokhimiya, Vol. 70, No. 2, 2005, pp. 197–202.Original Russian Text Copyright © 2005 by Mokhova, Khailova.This revised version was published online in April 2005 with corrections to the post codes.