长牡蛎Wnt5b基因的鉴定及在黑色素生成中的作用

Wnt基因家族编码一类保守的分泌型糖蛋白,广泛参与细胞分化与增殖、细胞运动、凋亡、免疫调控以及色素生成等生物学过程。本研究鉴定了一个长牡蛎Wnt家族基因——CgWnt5b,比较了黑、白壳色长牡蛎中CgWnt5b转录水平的表达情况,并对其在调控黑色素生成方面的功能进行初步研究。研究结果显示,CgWnt5b编码序列全长1116 bp,编码371个氨基酸。生物信息学分析预测CgWnt5b蛋白相对分子量为42 KDa,等电点为9.28。对CgWnt5b与哺乳动物Wnt基因家族19个成员进行系统发育分析,确定了CgWnt5b为哺乳动物Wnt5a和Wnt5b的同源基因。荧光实时定量PCR检测长牡蛎外套膜、鳃、消化腺、闭壳肌、性腺、唇瓣以及血淋巴组织中CgWnt5b的mRNA分布,结果显示,CgWnt5b在外套膜、闭壳肌和唇瓣组织中的相对表达水平较高,在消化腺、鳃、血淋巴和性腺组织中相对表达量较低。黑壳长牡蛎中CgWnt5b mRNA表达量显著低于白壳长牡蛎,且siRNA敲降CgWnt5b的表达后,多个黑色素生成相关酪氨酸酶基因的表达显著上调。综上,本研究在长牡蛎中鉴定了一个Wnt家族成员CgWnt5b,可能通过抑制黑色素生成关键基因TYR的表达抑制长牡蛎中黑色素的生成。     

Nickel isotope fractionation during laterite Ni ore smelting and refining: Implications for tracing the sources of Ni in smelter-affected soils

Nickel isotope ratios were measured in ores, fly ash, slags and FeNi samples from two metallurgical plants located in the Goiás State, Brazil (Barro Alto, Niquelândia). This allowed investigating the mass-dependent fractionation of Ni isotopes during the Ni-laterite ore smelting and refining. Feeding material exhibits a large range of δ⁶⁰Ni values (from 0.02 ± 0.10‰ to 0.20 ± 0.05‰, n = 7), explained by the diversity of Ni-bearing phases, and the average of δ⁶⁰Ni_{feeding materials} was found equal to 0.08 ± 0.08‰ (2SD, n = 7). Both δ⁶⁰Ni values of fly ash (δ⁶⁰Ni = 0.07 ± 0.07‰, n = 10) and final FeNi produced (0.05 ± 0.02‰, n = 2) were not significantly different from the feeding materials ones. These values are consistent with the very high production yield of the factories. However, smelting slags present the heaviest δ⁶⁰Ni values of all the smelter samples, with δ⁶⁰Ni ranging from 0.11 ± 0.05‰ to 0.27 ± 0.05‰ (n = 8). Soils were also collected near and far from the Niquelândia metallurgical plant, to evaluate the potential of Ni isotopes for tracing the natural vs anthropogenic Ni in soils. The Ni isotopic composition of the non-impacted topsoils developed on ultramafic rocks ranges from −0.26 ± 0.09‰ to −0.04 ± 0.05‰ (n = 20). On the contrary, the Ni isotopic composition of the non-ultramafic topsoils, collected close to the plant, exhibit a large variation of δ⁶⁰Ni, ranging from −0.19 ± 0.13‰ up to 0.10 ± 0.05‰ (n = 4). This slight but significant enrichment in heavy isotopes highlights the potential impact of smelting activity in the surrounding area, as well as the potential of Ni isotopes for discerning anthropogenic samples (heavier δ⁶⁰Ni values) from natural ones (lighter δ⁶⁰Ni values). However, given the global range of published δ⁶⁰Ni values (from −1.03 to 2.5‰) and more particularly those associated to natural weathering of ultramafic rocks (from −0.61 to 0.32‰), the use of Ni isotopes for tracing environmental contamination from smelters will remain challenging.     

不同精制方法对鱿鱼（Ommastrephes bartrami）墨黑色素的微观形态、光谱特征及金属元素的影响

采用扫描电镜、光谱分析和元素分析法,进行了胃蛋白酶水解法、高速离心法和酸水解法精制的鱿鱼墨黑色素颗粒形态、紫外可见光谱、红外光谱及金属元素含量的研究。结果表明,酸水解法破坏了鱿鱼墨黑色素的颗粒结构;三种不同精制方法所得的鱿鱼墨黑色素在220nm附近均有强吸收。胃蛋白酶水解和酸水解法有效切除黑色素的氨基基团,但酸水解法下...     

产胞外黑色素海洋菌的鉴定及发酵条件优化

从山东近海海域沉积物的微生物筛选中获得了一株高产胞外黑色素的菌株QJNY82,通过对该菌株菌落形态特征观察、生理生化检验及16S rDNA序列测定,初步确定该菌株为发光杆菌属细菌.通过对菌株QJNY82的胞外产物进行红外光谱分析,得到该细菌的胞外产物为黑色素,纯化后的黑色素的红外光谱图与标准黑色素（SIGMA）的红外光谱图一致,且该细菌所产黑色素的理化性质与标准黑色素具有一致性.该海洋菌株具有产黑色素速度快,产量高且不需要L-酪氨酸诱导的特点,而且经L-酪氨酸诱导后,可显著提高菌株QJNY82胞外黑色素的产量,在L-酪氨酸的含量为0.75 g/dm3、盐度为30、pH值为8.0、温度为28℃的条件下,菌株黑色素产量是未加L-酪氨酸的3.5倍,其产量达1.325g/dm3.鉴于菌株QJNY82具有产黑色素的特性,将成为发光杆菌属的一个新的菌种资源.     

白刺参，浅青刺参，深青刺参和紫刺参色素组成及黑色素含量的比较分析

Sea cucumber, Apostichopus japonicus(Selenka), is a commercially important marine species in China. Among the differently colored varieties sold in China, white and purple sea cucumbers have the greatest appeal to consumers. Identification of the pigments that may contribute to the formation of different color morphs of sea cucumbers will provide a scientific basis for improving the cultivability of desirable color morphs. In this study,sea cucumbers were divided into four categories according to their body color: white, light green, dark green, and purple. The pigment composition and contents in the four groups were analyzed by high performance liquid chromatography(HPLC). The results show that the pigment contents differed significantly among the white, lightgreen, dark-green, and purple sea cucumbers, and there were fewer types of pigments in white sea cucumber than in the other color morphs. The only pigments detected in white sea cucumbers were guanine and pteroic acid.Guanine and pteroic acid are structural colors, and they were also detected in light-green, dark-green, and purple sea cucumbers. Every pigment detected, except for pteroic acid, was present at a higher concentration in purple morphs than in the other color morphs. The biological color pigments melanin, astaxanthin, β-carotene, and lutein were detected in light-green, dark-green, and purple sea cucumbers. While progesterone and lycopene,which are also biological color pigments, were not detected in any of the color morphs. Melanin was the major pigment contributing to body color, and its concentration increased with deepening color of the sea cucumber body. Transmission electron microscopy analyses revealed that white sea cucumbers had the fewest epidermal melanocytes in the body wall, and their melanocytes contained fewer melanosomes as well as non-pigmented pre-melanosomes. Sea cucumbers with deeper body colors contained more melanin granules. In the body wall of dark-green and purple sea cucumbers, melanin granules were secreted out of the cell. The results of this study provide evidence for the main factors responsible for differences in coloration among white, light-green, darkgreen, and purple sea cucumbers, and also provide the foundation for further research on the formation of body color in sea cucumber, A. japonicus.     

The Decline of Competition: The emergence of a duopoly in the Australian sugar refinery sector, 1841–1915

The late nineteenth and early twentieth centuries saw the emergence of large single-product organisations whose expansion was stimulated by a desire to control a substantial part of a particular product market. An Australian example is the Colonial Sugar Refining Company Ltd (CSR). By 1915, the company accounted for 90 per cent of refined sugar production in Australia. CSR had achieved this control over the Australian refined sugar market by using underselling, strategies that denied its competitors outlets for their refined sugars and amalgamations with rivals. After 1890, however, CSR was interested in sharing the Australian market for refined sugar with its rivals by forming spheres of influence in marketing areas or adopting production quotas, although the Queensland National Bank was the only organisation to successfully negotiate annual market share agreements with CSR in the 1900s. These agreements ensured the survival of its Millaquin Refinery. Other competitors eventually exited the industry. In 1915, the Commonwealth government institutionalised this duopoly in the sugar refining sector by agreements with CSR and the Queensland government, and allowed it to continue until deregulation of the Australian sugar refining sector in the early 1990s.     

深海细菌Pseudomonas sp.bIp-2黑色素生成条件研究及其hppD基因的克隆

从大西洋深海沉积物中分离得到1株产黑色素细菌b Ip-2,16S rRNA基因序列分析表明该菌隶属于假单胞菌属（Pseudomonas）,与施氏假单胞菌（P.stutzeri）有最高的16S rRNA基因序列相似性（99%）.进一步对该菌的生长及产黑色素条件进行了初步研究,结果表明b Ip-2在稳定生长期后期才开始大量积累黑色素;b Ip-2可在4～37℃的温度范围下生长,最适生长温度为37℃,在28℃具有最高的黑色素产量;b Ip-2生长的pH范围偏碱性,在p H=7的最适生长条件下黑色素产量最高,同时碱性环境有利于黑色素的快速形成;b Ip-2可以在1%～9%的NaCl浓度范围中生长,在5%的NaCl浓度下具有最高的黑色素产量;低浓度的Fe^2＋（0.05 mmol/dm^3）可以提高黑色素的产量,Fe^2＋浓度≥0.20 mmol/dm^3时明显抑制黑色素的生成,而Fe^2＋浓度为1.00 mmol/dm^3时可完全抑制菌株的生长;低浓度的Cu^2＋（0.1 mmol/dm^3）对黑色素的产量无明显影响,Cu^2＋浓度≥0.5mmol/dm^3时可抑制黑色素的生成,而Cu^2＋浓度超过1.0 mmol/dm^3时可完全抑制菌株的生长.通过PCR扩增获得菌株b Ip-2的对羟苯基丙酮酸双加氧酶（hydroxyphenylpyruvic acid dioxygenase,HPPD）全长基因（1087 bp）,分析发现其编码的HPPD蛋白序列（含361氨基酸）与来自菌株Pseudomonas stutzeri RCH2的HPPD蛋白序列具有最高的同源性（序列相似性为97%）.这些结果为深入研究该细菌所产黑色素的合成,应用及生态作用奠定了基础.     

酸碱粗提配合超声破碎法制取可溶性长牡蛎(Crassostrea gigas)黑色素的研究

虽然牡蛎黑色素已经提取鉴定成功,但因为大部分实验需要用到黑色素的水溶液,而天然黑色素不溶于水,致使天然长牡蛎(Crassostrea gigas)黑色素的功效验证工作难以开展。本课题组探究总结出一套有效方法,对可溶性长牡蛎黑色素进行制取。首先选用酸碱法,从黑色长牡蛎中提取出天然长牡蛎黑色素,将其溶于氢氧化钠溶液中,利用超声细胞破碎仪进行处理,用盐酸中和至中性,离心取上清并干燥后得到一种黑色可溶性固体;再将该黑色固体分别用激光粒度测试仪、红外光谱仪和紫外光谱扫描仪进行检验后,发现天然长牡蛎黑色素经过超声降解处理后,颗粒粒度大幅下降,其溶解性大幅提高。红外光谱则显示出1630nm左右有明显的峰值,说明超声降解处理并未破坏真黑色素吲哚环等官能团;紫外吸光图谱的比较则显示天然长牡蛎黑色素破碎后,吸收峰仍然出现在210nm左右,但吸收值明显升高,说明超声降解法可以将难溶于水的天然长牡蛎黑色素降解为可溶于水的小颗粒可溶性黑色素,从而提高了紫外吸收值。可溶性牡蛎黑色素的制备成功对推动牡蛎黑色素的功能研究具有重要意义。     

Anwendung von biologischen Testverfahren an Abwässern der Textilindustrie

Testing Effluents of the Textile Refining Industry with Biological Methods The environmental problems caused by the manufacture of finished textiles involve a long chain of individual processes. This “textile chain” includes very diverse enterprises of varied size and structure. The textile refiners occupy a key position in the “textile chain”. On the one hand, this is due to their use of an obscurely large number of chemicals which can end up in the wastewater as well as in the textile products. On the other hand, this key role of the textile refining industry is based on their central position between the preproduction stage and the consumers. This study dealt with the textile refining industry's wastewater. As measured by volume and contents of its wastewater, this industry can be counted among the major industrial plants which discharge into municipal wastewater treatment plants. German wastewater legislation includes the provision that substances which are toxic, persistent, capable of accumulating, carcinogenic, fetotoxic or mutagenic be kept out of natural waters as well as technically possible (Wasserhaushaltsgesetz WHG). Several biotest methods for examining the effect of the substances contained in the wastewater were incorporated into the appendix of the German wastewater regulation (Rahmenabwasser-Verwaltungsvorschrift based on § 7a WHG). The aim of this study was to show, with the aid of biotest methods, how strongly the wastewater of textile refining companies is polluted as compared to other known industrial branches and to what degree the pollution of these wastewaters is eliminated by the treatment in wastewater treatment plants. Finally, we experimented to find out which biotest methods were suited for the examination of these wastewaters. The study's results show that the ecotoxicity of the textile refining industry's wastewater was only extraordinary high in isolated cases as compared to other examined branches of industry. The textile wastewaters exhibit values of G_L = 3 to G_L = 96 in the luminescent bacteria test, G_D = 1 to G_D = 192 (with one exception of G_D > 30000) in the daphnia test and G_F < = 2 to G_F = 32 in the fish test. It turned out though, that a large number of the samples from the textile refining companies (27%) reacted mutagenically in the Ames test in their native state. Consecutive tests for chromosomal aberrations (V79 hamster cell test) also showed mutagenic potential in five out of nine native samples. The employed testing methods with fish, daphnia and luminescent bacteria demonstrate a higher sensitivity of the luminescent bacteria and/or the daphnia as opposed to the fish in most cases. As the fish test is controversial anyway on the grounds of animal protection, a replacement of the fish test by these other tests should be aimed at: on account of the different end points of the luminescent bacteria and the daphnia test, a combination of these tests appears most sensible.