Effects of irradiation on neuropeptide expression in rat salivary gland and spinal cord

Summary It is well-known that a large number of factors can influence the expression of neuropeptides in the nervous system. In the present study, the effects of unilateral and bilateral irradiation to the rat head and neck on the expression of neuropeptides in the innervation of the submandibular gland and in the ganglionic cells of the submandibular ganglion was examined ten days and six months after treatment. Antisera directed against enkephalin and bombesin and immunohistochemical methods were used. The effects of bilateral irradiation on the staining pattern of various neuropeptides in the cervical spinal cord were also studied. In the submandibular gland and in the submandibular ganglionic cells, there was a markedly increased neuropeptide expression ten days after bilateral treatment, as seen after staining with both antisera used, while no changes occurred after unilateral treatment. Six months after treatment, the pattern of neuropeptide expression in the submandibular gland/ganglion corresponded to that seen in controls. Irradiation did not lead to any changes in the staining pattern of neuropeptides in the spinal cord. The observations show that there is a great complexity in the susceptibility of nervous tissues to radiotherapy with respect to influences on the expression of neuropeptides.     

ISOLATION AND CHARACTERIZATION OF PHASE-SPECIFIC COMPLEMENTARY DNAs FROM SPOROPHYTES AND GAMETOPHYTES OF PORPHYRA PURPUREA (RHODOPHYTA) USING SUBTRACTED COMPLEMENTARY DNA LIBRARIES1

The red alga Porphyra purpurea (Roth) C. Agardh has a life cycle that alternates between shell-boring, filamentous sporophytes and free-living, foliose gametophytes. The significant morphological differences between these two phases suggest that many genes should be developmentally regulated and expressed in a phase-specific manner. In this study, we prepared and screened subtracted complementary DNA (cDNA) libraries specific for the sporophyte and gametophyte of P. purpurea. This involved the construction of cDNA libraries from each phase, followed by the removal of common clones through subtractive hybridization. Sampling of the subtracted libraries indicated that 8–10% of the recombinant colonies in each library were specific for the appropriate phase. Of 20 putative phase-specific cDNAs selected from each subtracted library, eight unique clones were obtained for the sporophyte and seven for the gametophyte. After confirming their phase-specificities by hybridization to gametophyte and sporophyte messenger RNA, these 15 phase-specific cDNAs were sequenced, and the deduced amino acid sequences were used to search protein databanks. Two proteins encoded by the sporophyte-specific cDNAs and two by the gametophyte-specific cDNAs were identified by their similarity to databank entries.     

Protein synthesis is not required for the inhibitory effect of selenite on cell colony formation and RNA synthesis

Selenite has been shown to undergo intracellular metabolism that results in its conversion to other low molecular weight Secontaining species and also to its incorporation into a selenocysteine residue in selenoprotein. In order to investigate whether the incorporation into protein is required for the cytotoxic effects of selenite, we have examined whether inhibition of protein synthesis prevents the inhibitory effect of selenite on the ability of cells to form colonies or to synthesize RNA. We have found that treatment of HeLa cells with cycloheximide inhibited protein synthesis by >90% but had no effect on the inhibitory effect of selenite on cell colony formation or RNA synthesis. Since protein synthesis is not necessary for these cytotoxic effects of selenite they are unlikely to result from an increase in the synthesis of selenoproteins.     

The last 140 ka in the Afro-Asian arid/semi-arid transitional zone

During the last ten years, a great number of geological or other proxy records provided radiocarbon, U/Th or TL dated information on the past climatic oscillations in the arid/semi-arid zones extending between the Atlantic and the Pacific. Comparisons and a synthetic view of these results can now be attempted for the last 140 ka, and compared with global changes, as registered in oceanic or ice cores, and with palaeomonsson models: for this purpose, wide spatial and time scales have to be used. On the whole, arid/humid alternations roughly fit global changes, cold phases corresponding to an extension of the arid areas to the South, warm phases corresponding to the shrinking of the same to the North.

The last interglacial is associated with an increase of precipitation throughout the area considered. Isotopic stage 4 brings no evidence whatsoever of humid conditions. Two wetter episodes are registrated during stage 3. A major rainfall decrease is everywhere associated with the Last Glacial Maximum (21-15 ka in most regions), the arid or semi-arid zones extending several hundred kilometers southwards, relative to the present-day pattern. The two abrupt deglaciation steps and the Younger Dryas are recorded in all of the most sensitive regions, at the margins of the present-day monsoonal range. During the Holocene, the precipitation increases everywhere (by 100–400 mn, relative to the present-day values), the optimum being at 8.5-6.5 ka. A climatic deterioration follows with an irregular pattern of dry/wet episodes, according to the different geographic conditions. The humid phase terminates at 3.5-3 ka in the whole transitional zone.

The major southward shift of the monsoonal precipitation range since its optimum, some 8000 years ago, fits the astronomical neoglacial trend and may possibly be correlated with past stage 5d, although its rapidity and spatial importance may just reflect one of the sharp successive cold/warm variations registered by GRIP during the whole stage 5.     

旋毛虫肌幼虫ES抗原的基因克隆及高效表达

作者对编码旋毛虫肌幼虫ES抗原的部分结构基因进行了克隆、鉴定和表达。用RNA PCR技术直接从旋毛虫肌幼虫总RNA中反转录并扩增出0．7kh的靶DNA,酶切分析后将其克隆到融合表达载体pEx3lC中。SDS—PAGE电泳表明,含重组子的大肠杆菌能够表达出一分子量为37kDa的融合蛋白(P37),后者占菌体总蛋白的22%以上,并以包含体形式存在于菌体中。经对纯化后表达蛋白的ELlSA检测,证明它能被猪旋毛虫病阳性血清和抗旋毛虫单克隆抗体识别。研究结果揭示,重组蛋白P37对于研制旋毛虫病诊断抗原和免疫抗原具有潜在的应用价值。     

双价外壳蛋白基因植物表达载体的构建及马铃薯转基因植株的鉴定

在克隆了马铃薯Ｘ病毒（ＰＶＸ）、马铃薯Ｙ 病毒（ＰＶＹ）和马铃薯卷叶病毒（ＰＬＲＶ）的外壳蛋白基因的基础上,构建同时包含ＰＶＸ和ＰＶＹ 与ＰＶＹ 和ＰＬＲＶ 两个外壳蛋白基因植物表达框架的表达载体,通过农杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ）介导转化烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）和生产上常用的几个马铃薯（Ｓｏｌａｎｕｍ ｔｕｂｅｒｏｓｕｍ ）优良品种：“Ｆａｖｏｒｉｔａ”、“虎头”、“克４”。经ＰＣＲ检测证明外源基因已整合到植物的染色体上,得到批量转基因植株。在转ＰＶＸ＋ＰＶＹ 外壳蛋白基因的烟草上接种ＰＶＸ （５ μｇ／ｍ Ｌ）、ＰＶＹ（２０ μｇ／ｍ Ｌ）病毒,得到有一定抗性的植株     

花粉高尔基囊泡与牛脑微管的体外结合

把从榛木（Ｃｏｒｙｌｕｓａｖｅｌｌａｎａ Ｌ．）花粉中分离得到的高尔基囊泡与经高度纯化并聚合好的牛脑微管进行体外组合,然后于１．５ ｍ ｏｌ／Ｌ的蔗糖层上进行超离心,对其沉淀物进行ＳＤＳ－聚丙烯酰胺凝胶电泳和电镜负染。结果表明,花粉高尔基囊泡可以结合到牛脑微管上,证明植物花粉的高尔基囊泡与动物细胞的某些细胞器一样,也与细胞骨架的主要组成之一——微管具有结构上的紧密联系。花粉高尔基囊泡与牛脑微管的体外结合能力,受１０ ｍ ｍ ｏｌ／ＬＡＴＰ和０．５ ｍ ｏｌ／ＬＫＣｌ的影响,但不受５ ｍ ｍ ｏｌ／Ｌ ＡＭＰ－ＰＮＰ的影响,说明两者结合可能是通过高尔基囊泡表面与ＡＴＰ有关的某种外周膜蛋白来完成的。     

几种濒危植物及其近缘类群总DNA的提取与鉴定

用低ｐＨ 介质,高盐沉淀蛋白质方法成功地从银杉（Ｃａｔｈａｙａ ａｒｇｙｒｏｐｈｙｌｌａ Ｃｈｕｎ ｅｔＫｕａｎｇ）、矮牡丹（Ｐａｅｏｎｉａ ｓｕｆｆｒｕｔｉｃｏｓａ ｖａｒ． ｓｐｏｎｔａｎｅａ）、南川升麻（Ｃｉｍ ｉｃｉｆｕｇａ ｎａｎｃｈｕａｎｅｎｓｉｓＨｓｉａｏ）、裂叶沙参（Ａｄｅｎｏｐｈｏｒａ ｌｏｂｏｐｈｙｌｌａ）的同属种泡沙参（Ａ． ｐｏｔａｎｉｎｉｉ）等植物中提取和部分纯化了细胞总ＤＮＡ,并对其产率、质量和纯度作了鉴定。此方法的关键是用了一个低ｐＨ提取介质,它能有效防止组织破碎及沉淀大量材料时的电离化作用及酚化合物的进一步氧化。所得ＤＮＡ 不需经氯化铯梯度离心或柱层析,直接可用于限制性片断长度多态性（ＲＦＬＰ）及随机扩增的ＤＮＡ多态性（ＲＡＰＤ）等分子水平的遗传标记。为检测濒危植物的遗传多样性提供了一套迅速、简便和可靠的技术方案     

Weak antioxidant defenses make the heart a target for damage in copper-deficient rats

Copper deficiency causes more salient pathologic changes in the heart than in the liver of rats. Although oxidative stress has been implicated in copper deficiency-induced pathogenesis, little is known about the selective toxicity to the heart. Therefore, we examined the relationship between the severity of copper deficiency-induced oxidative damage and the capacity of antioxidant defense in heart and liver to investigate a possible mechanism for the selective cardiotoxicity. Weanling rats were fed a purified diet deficient in copper (0.4 μg/g diet) or one containing adequate copper (6.0 μg/g diet) for 4 weeks. Copper deficiency induced a 2-fold increase in lipid peroxidation in the heart (thiobarbituric assay) but did not alter peroxidation in the liver. The antioxidant enzymatic activities of superoxide dismutase, catalase, and glutathione peroxidase were, respectively, 3-, 50- and 1.5-fold lower in the heart than in the liver, although these enzymatic activities were depressed in both organs by copper deficiency. In addition, the activity of glutathione reductase was 4 times lower in the heart than in the liver. The data suggest that a weak antioxidant defense system in the heart is responsible for the relatively high degree of oxidative damage in copper-deficient hearts.