Capillary electrophoretic study on UDP-sugars in cells

Glucose is an important regulator of cell growth and metabolism. Uridine diphosphate sugars (UDP-sugars), as the intermediate products of metabolism, play pivotal roles as precursors in the synthesis of complex carbohydrates and glycolipids as well as lectose. It is very important to study their metabolism in cells in clinical biochemistry. A capillary electrophoretic method has been developed for the analysis of UDP-sugars and nucleotides, By using an uncoated capillary (70cm x 50 mu m) and 20 mmol/L borax buffer (pH 9), 4 important UDP-sugars can be analyzed in 15 min at 22 kV with satisfactory precision and sensitivity. The developed method has been applied to analyze UDP-sugars concentrations in lymphocytes, fibroblasts and mesangial cells, and the results show it not only is much better than HPLC method, but also can be used to measure the energy charge of cells.

Sugars-grafted aliphatic biodegradable poly(L-lactide-co-carbonate)s by click reaction and their specific interaction with lectin molecules

A novel biodegradable aliphatic poly(L-lactide-co-carbonate) bearing pendant acetylene groups was successfully prepared by ring-opening copolymerization of L-lactide (LA) with 5-methyl-5-propargyloxycarbonyl-1,3-dioxan-2-one (PC) in the presence of benzyl alcohol as initiator with ZnEt2 as catalyst in bulk at 100 degrees C and subsequently used for grafting 2-azidoethyl beta-D-glucopyranoside and 2-azidoethyl beta-lactoside by the typical "click reaction," that is Cu(I)-catalyzed cycloaddition of azide and alkyne. The density of acetylene groups in the copolymer can be tailored by the molar ratio of PC to LA during the copolymerization. The aliphatic copolymers grafted with sugars showed low cytotoxicity to L929 cells, improved hydrophilic properties and specific recognition and binding ability with lectins, that is Concanavalin A (Con A) and Ricinus communis agglutinin (RCA). Therefore, this kind of sugar-grafted copolymer could be a good candidate in variety of biomedical applications.

葡萄浆果成熟与延迟采收期间糖酸变化特点的研究

连续2年对“京秀”ד亚历山大”（编号97-2）杂交后代10个单株和“京秀”ד香妃”（编号97-3）杂交后代10个单株成熟期和延迟采收期的糖酸组分含量进行测定，以了解葡萄浆果延迟采收内在品质的变化差异。第二年 对可长期挂果的亲本“京秀”、“亚历山大”和杂交后代单株“97-2-23”、“97-2-24”成熟期和延迟采收期糖酸组分动态变化进行测定，以了解葡萄浆果糖酸品质延迟采收期的动态变化规律。同时对以上四个单株的浆果中蔗糖代谢相关酶活性和浆果呼吸强度进行测定，以对不同蔗糖表现型的产生原因进行探讨。研究表明： 葡萄延迟采收，浆果内可溶性糖变化可分为积累型和稳定型两种类型。延迟采收3周，糖积累型的8个单株浆果 可溶性糖显著积累，而稳定型的12个单株的糖与成熟时不存在显著差异。浆果主要积累果糖和葡萄糖，这两种糖的 动态变化趋势基本一致，但果糖变化幅度较大。两种类型的有机酸含量在延迟采收期间变化不大，没有显著差异。 糖积累型的“京秀”和“97-2-23”延迟采收期间各蔗糖分解酶（酸性转化酶、中性转化酶、蔗糖合成酶分解方 向）活性持续升高，但糖稳定型的“亚历山大”和“97-2-24”延迟采收第3周时急剧降低。此外，果实成熟和延迟 采收期间两种糖表现型浆果的呼吸速率也存在差异。浆果的蔗糖分解酶活性和呼吸速率存在的差异可能是两种类型 延迟采收期间糖含量存在差异的重要原因。

Effect of sugar type on the survival of frozen-thawed rhesus monkey (Macaca mulatta) sperm

Sperm-freezing extenders supplemented with sugar or a combination of different sugars are widely used for the cryopreservation of nonhuman primate spermatozoa. Understanding which sugar or combination of sugars offers the highest level of cryoprotection w

Cytochemical changes in the developmental process of Nostoc sphaeroides (cyanobacterium)

There are several apparent developmental stages in the life cycle of Nostoc sphaeroides Kutzing, an edible cyanobacterium found mainly in paddy fields in central China. The cytochemical changes in developmental stages such as hormogonia, aseriate stage, filamentous stage and colony in N. sphaeroides were examined using fluorescent staining and colorimetric methods. The staining of acidic and sulfated polysaccharides increased with development when hormogonia were used as the starting point. Acidic polysaccharides (AP) were most abundant at the aseriate stage and then decreased, while the sulfated polysaccharides (SP) were highest at the colony stage. Quantitatively, along the developmental process from hormogonia to colony, total carbohydrates first increased, then became stable, and then reached their highest level at the colony stage, while reducing sugars were highest at the hormogonia stage and then decreased sharply once development began. SP were not detectable in the hot water soluble polysaccharides (HWSP), and hormogonia had the lowest content of AP, while old colonies had the highest. The AP content of the aseriate stage, filamentous stage and young colony stage were very similar. The evolutionary relationships reflected in the developmental stages of N. sphaeroides are discussed.

UV-B-induced Oxidative Damage and Protective Role of Exopolysaccharides in Desert Cyanobacterium Microcoleus vaginatus

UV-B-induced oxidative damage and the protective effect of exopolysaccharides (EPS) in Microcoleus vaginatus, a cyanobacterium isolated from desert crust, were investigated. After being irradiated with UV-B radiation, photosynthetic activity (Fv/Fm), cellular total carbohydrates, EPS and sucrose production of irradiated cells decreased, while reducing sugars, reactive oxygen species (ROS) generation, malondialdehyde (MDA) production and DNA strand breaks increased significantly. However, when pretreated with 100 mg/L exogenous EPS, EPS production in the culture medium of UV-B stressed cells decreased significantly; Fv/Fm, cellular total carbohydrates, reducing sugars and sucrose synthase (SS) activity of irradiated cells increased significantly, while ROS generation, MDA production and DNA strand breaks of irradiated cells decreased significantly. The results suggested that EPS exhibited a significant protective effect on DNA strand breaks and lipid peroxidation by effectively eliminating ROS induced by UV-B radiation in M. vaginatus.

Effects of salt stress on carbohydrate metabolism in desert soil alga Microcoleus vaginatus Gom.

The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.

Photoregulated or energy dependent process of hormogonia differentiation in Nostoc sphaeroides Kutzing (Cyanobacterium)

Hormogonium, which was thought to play an important role in the dispersal and survival of these microorganisms in their natural habitats, is a distinguishable developmental stage of heterocystous cyanobacteria. The present study examined the effects of different light conditions and sugars on the differentiation of Nostoc sphaeroides Kutzing to the hormogonia stage. Results showed that differentiation of hormogonia was light dependent in the absence of sugar, but that close to 100% of cyanobacteria differentiated to hormogonia in the presence of glucose or sucrose, irrespective of the light conditions. This differentiation was inhibited, even in the presence of sugars, upon application of an inhibitor of respiration. Following the testing of different sugars, the effects of different lights were examined. It was found that 5 10 μ mol.m(-2)• s(-1) photon flux density was optimal for hormogonia differentiation. One hundred percent differentiation was obtained with white light irradiation, in contrast with irradiation with green light (80% differentiation) and red light (0-10% differentiation). Although they showed different efficiencies in inducing hormogonia differentiation in N. sphaeroides, the green and red radiation did not display antagonistic effects. When the additional aspect of time dependence was investigated through the application of different light radiations and an inhibitor of protein synthesis, it was found that the initial 6 h of the differentiation process was crucial for hormogonia differentiation. Taken together, these results show that hormogonia differentiation in N. sphaeroides is either a photoregulated or an energy dependent process.

Studies on polysaccharides from three edible species of Nostoc (Cyanobacteria) with different colony morphologies: Comparison of monosaccharide compositions and viscosities of polysaccharides from field colonies and suspension cultures

Hot water-soluble polysaccharides woe extracted from field colonies and suspension cultures of Nostoc commune Vaucher, Nostoc flagelliforme Berkeley et Curtis, and Nostoc sphaeroides Kutzing. Excreted extracellular polymeric substances (EPS) were isolated from the media in which the suspension cultures were grown. The main monosaccharides of the field colony polysaccharides from the three species were glucose, xylose, and galactose, with an approximate ratio of 2:1:1. Mannose was also present, but the levels varied among the species, and arabinose appeared only in N. flagelliforme. The compositions of the cellular polysaccharides and EPS from suspension cultures were more complicated than those of the field samples and varied among the different species. The polysaccharides from the cultures of N. flagelliforme had a relatively simple composition consisting of mannose, galactose, glucose, and glucuronic acid, but no xylose, as was found in the field colony polysaccharides. The polysaccharides from cultures of N. sphaeroides contained glucose (the major component), rhamnose, fucose, xylose, mannose, and galactose. These same sugars were present in the polysaccharides from cultures of N. commune, with xylose as the major component. Combined nitrogen in the media had no qualitative influence on the compositions of the cellular polysaccharides but affected those of the EPS of N. commune and N. flagelliforme. The EPS of N. sphaeroides had a very low fetal carbohydrate content and thus was not considered to be polysaccharide in nature. The field colony polysaccharides could be separated by anion exchange chromatography into neutral and acidic fractions having similar sugar compositions. Preliminary linkage analysis showed that 1) xylose, glucose, and galactose were 1-->4 linked, 2) mannose, galactose, and xylose occurred as terminal residues, and 3) branch points occurred in glucose as 1-->3,4 and 1-->3,6 linkages and in xylose as a 1-->3,4 linkage. The polymer preparations from field colonies had higher kinematic viscosities than those from corresponding suspension cultures. The high viscosities of the polymers suggested that they might DE suitable for industrial uses.

Kinetic study of hydrolysis of xylan and agricultural wastes with hot liquid water

We investigated the kinetics of hot liquid water (HLW) hydrolysis over a 60-min period using a self-designed setup. The reaction was performed within the range 160-220 °C, under reaction conditions of 4.0 MPa, a 1:20 solid:liquid ratio (g/mL), at 500 rpm stirring speed. Xylan was chosen as a model compound for hemicelluloses, and two kinds of agricultural wastes-rice straw and palm shell-were used as typical feedstocks representative of herbaceous and woody biomasses, respectively. The hydrolysis reactions for the three kinds of materials followed a first-order sequential kinetic model, and the hydrolysis activation energies were 65.58 kJ/mol for xylan, 68.76 kJ/mol for rice straw, and 95.19 kJ/mol for palm shell. The activation energies of sugar degradation were 147.21 kJ/mol for xylan, 47.08 kJ/mol for rice straw and 79.74 kJ/mol for palm shell. These differences may be due to differences in the composition and construction of the three kinds of materials. In order to reduce the decomposition of sugars, the hydrolysis time of biomasses such as rice straw and palm shell should be strictly controlled.

土壤氨基酸和氨基糖聚合物的矿化过程研究

土壤中氨基酸和氨基糖是土壤有机氮的重要组成部分，对土壤氮素供给和土壤碳、氮循环过程有重要贡献。研究氨基酸和氨基糖聚合物的矿化，对于减少氮素损失，提高氮肥利用率能够提供一定的理论依据。 当向土壤中(本试验为黑土)同时添加葡萄糖和(15NH4)2SO4时，在土壤微生物的作用下，(15NH4)2SO4会被用以合成土壤15N-氨基酸和氨基糖聚合物。新合成的这部分氨基酸和氨基糖聚合物与土壤中原有氨基酸和氨基糖聚合物的性质是否不同并且是否受到外源底物的调控。为了解决上述问题，本试验将采用Stanford and Smith的间歇好气矿化淋洗培养法，结合高效液相色谱/质谱、气相色谱/质谱联机技术(HPLC/MS，GC/MS)跟踪测定土壤中15N-氨基酸和氨基糖的同位素富集比例及其含量的变化，探讨土壤中新合成15N-氨基聚合物的矿化特征，通过研究添加葡萄糖、玉米秸秆和无机氮肥对土壤中新合成15N-氨基聚合物矿化过程的影响以及对有机氮聚合物解聚的动力－酶活性的影响，从而阐明土壤氨基聚合物矿化的碳源营养调控机制和氮源反馈调节机制及其解聚机理。研究结果表明： 1. 土壤中新合成的氨基酸和氨基糖聚合物与土壤原有氨基酸和氨基糖聚合物相比具有较高的循环速率，并且不同种氨基酸和氨基糖也分别表现出不同的矿化特征。较高循环速率的存在，将为调控其矿化过程奠定基础，因为只有快速循环的氮素才能够被调控，而且调控新合成有机氮的矿化过程，可以不断地满足作物生长对氮素养分的需求。 2. 土壤中新合成氨基聚合物的矿化受到不同外源底物调控。其中碳源(葡萄糖和玉米秸秆)能够抑制氨基聚合物矿化，但是活性碳源葡萄糖的抑制程度高于活性较低的碳源玉米秸秆，表明氨基聚合物矿化受到不同碳源活性调控。不同浓度以及不同形式氮源也能够调控土壤氨基聚合物的矿化，并且适量氮肥的加入能够抑制土壤中新合成氨基聚合物的矿化，存在氮肥的反馈抑制机制。 3. 土壤中蛋白酶、芳基酰胺酶和几丁质酶活性受到不同碳源和氮源的影响。其中碳源表现为促进作用，而氮源则表现为抑制作用。氮源对土壤酶活性的反馈抑制作用是控制土壤氮素转化的关键，而碳源只是起到维持土壤酶活性的作用。三种酶活性对外源底物的敏感性，将对于调控土壤氮素循环奠定一定的理论依据。 4. 不同处理酶活性与有机氮矿化之间表现出不同的相关性，说明酶与氮矿化之间的关系受到多方面因素影响。总体来看，蛋白酶、芳基酰胺酶和几丁质酶在水解土壤氨基酸和氨基糖聚合物的过程中起到重要作用，是有机氮聚合物重要的解聚酶。

外源底物对土壤氨基糖生物转化的影响

研究土壤碳、氮微生物转化过程和去向对于理解土壤有机质和肥力特性有重要意义，土壤氨基糖主要由微生物合成，且具有相对稳定性和异源性，所以跟踪微生物残留物氨基糖的动态变化是理解这个转化过程的有效手段。本试验采用室内模拟培养法，研究大豆叶和玉米秸在不同土壤中的微生物转化过程，通过土壤氨基糖组成和含量的动态变化来揭示土壤营养物质的转化机制，同时应用氨基糖特性来评价该过程中微生物的作用，为土壤碳，氮循环和截获的研究提供理论基础。土壤氨基糖在有机物料分解初期以合成为优势，后期以分解为优势，外源底物的数量和营养结构是影响有机物料在土壤中分解进程的主要因素，但外源底物的分解进程可以通过土壤和外加营养进行补偿和调节，充足和协调的营养供应可加速有机物料的分解进程。在底物的分解过程中，土壤氨基糖占有机质的比例和培养时间符合抛物线模型，并且不同底物下，该比例随培养时间加长逐渐趋近，直至重合，表明土壤有机质富积氨基糖的能力是相对稳定的。由此提出了稳定库—易变过渡库理论假设，氨基糖的变化主要在过渡库中，底物越丰富氨基糖过渡库的容量越大，真菌来源的氨基糖相对于细菌来源氨基糖对土壤活性碳、氮库的贡献也越大。