Comparison of the Soluble and Membrane-Bound Forms of the Puromycin-Sensitive Enkephalin-Degrading Aminopeptidases from Rat

Enkephalin degradation in brain has been shown to be catalyzed, in part, by a membrane-bound puromycin-sensitive aminopeptidase. A cytosolic puromycin-sensitive aminopeptidase with similar properties also has been described. The relationship between the soluble and membrane forms of the rat brain enzyme is investigated here. Both of these aminopeptidase forms were purified from rat brain and an antiserum was generated to the soluble enzyme. Each of the aminopeptidases is composed of a single polypeptide of molecular mass 100 kilodaltons as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography. The antisoluble aminopeptidase antiserum reacts with both enzyme forms on immunoblots and inhibits both with nearly identical inhibition curves. The isoelectric points (pI = 5.0) of both forms were shown to be identical. N-terminal sequencing yielded a common sequence (P-E-K-R-P-F-E-R-L-P-T-E-V-S-P-I-N-Y) for both enzyme forms, and peptide mapping yielded 26 peptides that also appeared identical between the two enzyme forms. Studies on the nature of the association of the membrane enzyme form with the cell membrane suggest that this enzyme form does not represent the soluble form trapped during the enzyme preparation. It is suggested that the membrane form of the puromycin-sensitive aminopeptidase is identical to the soluble enzyme and that it associates with the membrane by interactions with other integral membrane proteins.     

Characterization of Membrane-Bound Aminopeptidases from Rat Brain: Identification of the Enkephalin-Degrading Aminopeptidase

Rat brain aminopeptidase activity was solubilized from membranes by incubation with thiols. This novel procedure resulted in the release of the same two aminopeptidases (MI and MII) previously shown to be solubilized by the nonionic detergent Triton X-100. The solubilized aminopeptidases MI and MII were resolved by ion-exchange chromatography and further purified by hydroxylapatite chromatography. Aminopeptidase MI was shown to hydrolyze only the beta-naphthylamides of arginine and lysine whereas aminopeptidase MII exhibited a broad specificity with respect to amino acid beta-naphthylamides. Only aminopeptidase MII hydrolyzed Leu-enkephalin at a significant rate, indicating that this enzyme can account for the membrane-bound enkephalin aminopeptidase activity. The enkephalin-degrading aminopeptidase is potently inhibited by opioid (alpha-neo-endorphin and dynorphin) as well as nonopioid (substance P, somatostatin, and angiotensin I) peptides in the range of 0.2-2.0 microM. The regional distribution of aminopeptidases MI and MII in rat brain are rather different, with aminopeptidase MII distribution more closely paralleling the distribution of opiate receptors.     

红豆杉组织培养中褐变问题的研究

本研究针对红豆杉组织培养中经常遇到的褐变问题,采取多种方法进行试验,力求最大限度地避免或减轻褐变。结果表明,选择合适的外植体、培养基和培养条件是至关重要的,此外,抗氧化剂、金属离子螫合剂、热激处理和活性碳处理等方法可在一定程度上减轻褐变的程度。综合考虑各种因素,经过筛选可以获得生长良好的不发生褐变的组织。     

杨树的组织培养及其基因工程研究

杨树是林木基因工程的模式植物,在其组织培养过程中,试管苗的再生不仅与植株的基因型,年龄及组织的来源,状态等有关,还受许多外界因素如盐浓度及激素的种类与配比的影响,在杨树的转化过程中,DNA直接转移法和农杆菌介导法都有应用,但后者较为常用,组织培养及转化技术的日趋成熟,为杨树基因工程奠定了良好基础,本文对杨树组织培养,DNA转化方法及其基因工程进行了较为系统的概述。     

生姜组织培养污染率控制研究

为解决生姜组织培养过程中污染率较高的问题,就降低组织培养污染率的措施进行了研究。结果表明,在初代培养时采用采用剥离至茎尖仅剩3～4叶原基进行表面消毒,然后接种于含25mg／l～50gm/l的硫酸链霉素和硫酸卡那霉素混合液效果较好,在继代培养过程中培养基中抗生素浓度达到25～50mg／l,污染的培养基基本表现全无菌状态。     

Distribution of tripeptidyl-peptidase II in the central nervous system of rat

Tripeptidyl-peptidase II (TPP II) is a high molecular weight serine peptidase which removes tripeptides from a free N-terminus of longer peptides. Since it had previously been demonstrated that the enzyme can inactivate enkephalins and dynorphins in vitro by removing the N-terminal Tyr-Gly-Gly peptide, we wanted to see whether TPP II could be involved in this process also in vivo. Therefore, the localization of TPP II in different cerebral regions of rat was investigated by immunoblot analysis and activity measurements. It could be shown that TPP II is relatively evenly distributed in the central nervous system of rat. This indicates that the physiological role of the enzyme is probably not a specific degradation of enkephalins, but rather pertains to the general turnover of proteins.     

αγ-Enolase in the Rat: Ontogeny and Tissue Distribution

Abstract: The rat brain enolases are dimers composed of α and γ subunits. At pH 8.6 αγ-enolase seemed to be stable, and no evidence was found for the possible formation of αγ-enolase from αα-enolase and γγ-enolase in the course of rat brain homogenization. During ontogeny of the rat forebrain, αγ-enolase was formed before γγ-enolase. The half-maximal specific concentrations were reached at postnatal days 14 and 23, respectively. The distribution of αγ- and γγ-enolase in various rat brain areas was also investigated. In all areas both forms were present. In neuroendocrine tissues αγ-enolase was present at a much higher concentration than γγ-enolase. The ratio between γγ-enolase and αγ-enolase may be indicative of the degree of neuronal maturation, a conclusion further substantiated by the high ratio observed in cerebellum and the low ratio observed in olfactory bulbs, both compared with the ratio in forebrain.     

Tissue specification and intracellular distribution of actin isoforms inVicia faba L.

Summary Two dimensional gel electrophoresis of total cell protein extracts from not expanded, and primary leaves, petioles, and roots ofVicia faba resulted in four actin isoforms at 43 kDa with pI values from 5.9 to 6.05. In contrast to root extracts, in all leaf extracts an additional immunoreactive polypeptide with a molecular mass of 51 kDa and pI 5.75 was detected. This polypeptide was present in high amounts in protein extracts of purified chloroplasts, whereas no actin isoform at 43 kDa could be demonstrated. Compared to the tissue extracts, two actin isoforms at 43 kDa with pI values of 5.9 and 6.0 were enriched, when purified plasma membranes and the membranous fraction of vacuoles were analysed. In contrast, the soluble protein fraction of the plasma membrane preparation contained only two isoactins with pI values of 5.95 and 6.05 and a molecular mass of 43 kDa. These results indicate, that the four actin isoforms at 43 kDa detected in all examined tissues ofV. faba fulfill different functions at specific intracellular compartments, for example, the anchorage of actin microfilaments to membranes.Abbreviations BSA bovine serum albumin - BCIP 5-bromo-4-chloro-3-indolyl phosphate - DDM n-decyl -D-maltopyranoside - EDTA ethylenediamine-tetraacetic acid - HG n-hexyl -D-glucopyranoside - IEF isoelectrical focusing - MES morpholinoethanesulfonic acid - 2-ME 2 mercaptoethanol - NBT nitro blue tetrazolium - pCMB p-chloromercuribenzoic acid - PVP polyvinylpyrrolidone - Tris tris (hydroxymethyl) aminomethane     

金线莲组织培养中若干因素的研究

研究了影响金钱莲组培芽的增殖和营养器官分化的几个因素。结果表明:植物细胞分裂素及生长素的种类、浓度和配比对瓶苗茎发育有很大影响,在增殖期使用浓度为1.0mg/L BA、0.5mg/L IBA,器官分化生长期使用浓度为增殖期激素量的三分之一;瓶移植量10～15株;光照量显著影响植株生长期叶片生长,500lx较合适;活性炭明显提高诱导根的生长质量,添加浓度0.5％效果最好。     

油菜和玉米花粉粒中元素分布的研究

以油菜和玉米花粉为材料 ,应用扫描电镜及能谱仪技术对花粉中内外元素分布进行了初步研究。结果表明 :花粉粒萌发沟区与非萌发沟区 ,花粉壁与原生质内外的不同部位 ,Fe、Zn、Si、Al、Ca、P、S、Mg、K、Cl、Cu、Mn、Co及Ni1 4种元素组成有很大差异 ;两种花粉的萌发沟区都未检测出Fe元素 ,而非萌发沟区的Fe元素相对含量为 1 .1 2 %～ 3 68% ;花粉壁中比花粉原生质中更富含Si,Ca,K ,Cl等元素。本项研究为蜜源植物花粉的开发和利用提供了有价值的资料。     

Tissue distribution evaluation of stealth pH-sensitive liposomal cisplatin versus free cisplatin in Ehrlich tumor-bearing mice

Multidrug resistance and drug toxicity represent major obstacles to cancer chemotherapy. Drug delivery systems, such as liposomes, offer improved chemical stability of encapsulated drugs, enhanced accumulation in tumors and decreased toxicity. The aim of this study was to evaluate the tissue distribution of stealth pH-sensitive liposomes containing cisplatin (SpHL-CDDP), compared with free cisplatin (CDDP), in solid Ehrlich tumor-bearing mice. After administering a 6 mg/kg single intravenous bolus injection of either free radiolabeled cisplatin or SpHL containing radiolabeled cisplatin, blood and tissues were analyzed for cisplatin content by determining radioactivity using an automatic scintillation apparatus. The area under the CDDP concentration-time curve (AUC) obtained for blood after SpHL-CDDP administration was 2.1 fold larger when compared with free CDDP treatment. The longer circulation of SpHL-CDDP led to a higher tumor AUC, and the determination of the ratio between AUC in each tissue and that in blood (Kp) showed a higher accumulation of CDDP in SpHL-CDDP administrated tumors. The SpHL-CDDP was also significantly uptaken by the liver and spleen. The distribution of SpHL-CDDP in these organs was extensive, revealing a high extravasation of CDDP to the tissues. The SpHL-CDDP kidney uptake was also greater than that of free CDDP; however, the Kp value found was lower. This indicates that the SpHL-CDDP led to a reduction of CDDP retention by renal tissue. Thus, these results indicate that the SpHL-CDDP may indeed be useful in alleviating renal damage induced by CDDP and thus represents a promising delivery system for cancer treatment through CDDP.     

固定化细胞分布对动力学影响的模型化研究

通过对固定化Gluconobacter oxydans和Bacillus cereus的活细胞系统的研究,提出了固定化细胞不均匀分布模型,将此类分布与均匀分布(最劣分布模型)进行比较,分析了它们对固定化细胞内部的基质浓度分布、有效速率因子和选择率的影响,指出不均匀分布和均匀分布对于该固定化活细胞系统的动力学行为无显著影响,这一模型分析结果在实验中得以验证。通过无因次化分析,建立了适用于固定化生物催化剂动力学研究的模型方法。     

Preliminary Studies on the Geographic Distribution of Fagus in China

There are 11 species in Fagus, 5 of them are endemic to China. They are the major trees of deciduous broadleaved forest in Northern temperate region. This paper mainly deals with the relationship between the distribution of 5 species and 5 climatic indices. The results showed that the principal factor controlling the distribution of Fagus was the precipitation in growing season. The climatic change which resulted from Himalayan movement and the changes during pogt-glacial period and human disturbance also affect the contraction of the range of distribution.     

Tissue Distribution of Anidulafungin in Neonatal Rats

Anidulafungin, an echinocandin, is currently approved for treatment of fungal infections in adults. There is a high unmet medical need for treatment of fungal infections in neonatal patients, who may be at higher risk of infections involving bone, brain, and heart tissues. This in vivo preclinical study investigated anidulafungin distribution in plasma, bone, brain, and heart tissues in neonatal rats. Postnatal day (PND) 4 and PND 8 Fischer (F344/DuCrl) rats were dosed subcutaneously once with anidulafungin (10 mg/kg) or once daily for 5 days (PND 4–8). Plasma and tissue samples were collected and anidulafungin levels were measured by liquid chromatography‐tandem mass spectrometry. The mean plasma C_max and AUC_0‐24 values were consistent with single‐dose plasma pharmacokinetics (dose normalized) reported previously for adult rats. Observed bone concentrations were similar to plasma concentrations regardless of dosing duration, with bone‐to‐plasma concentration ratios of approximately 1.0. Heart concentrations were higher than plasma, with heart to plasma concentration ratios of 1.3‐ to 1.8‐fold. Brain concentrations were low after single dose, with brain‐to‐plasma concentration ratio of approximately 0.23, but increased to approximately 0.71 after 5 days of dosing. Tissue concentrations were nearly identical after single‐dose administration in both PND 4 and PND 8 animals, indicating that anidulafungin does not appear to differentially distribute in this period in neonatal rats. In conclusion, anidulafungin distributes to bone, brain, and heart tissues of neonatal rats; such results are supportive of further investigation of efficacy against infections involving bone, brain, and heart tissues.     

通过组织培养筛选小麦抗赤霉病突变体的研究

选用中抗赤霉病的春小麦品种和品系的花药进行离体培养,以小麦赤霉病29号菌株产生的致病培养滤液为选择剂,结合物理诱变处理,进行抗病鉴定,用赤霉病菌分生孢子直接接种在愈伤组织和再生植株筛选抗病突变体。在83块愈伤中有53块抗病。在11株的再生植株中有9株均比未经培养滤液处理的对照提高了抗病性,从中选出4株抗病性接近或超过“苏麦3号”品种。     

四川裂腹鱼脑型芳香化酶Cyp19b基因组织分布及温度对其表达水平的影响

为研究脑型芳香化酶基因Cyp19b在四川裂腹鱼(Schizothorax kozlovi)早期性别分化中的作用,采用RACE方法从四川裂腹鱼脑中扩增得到该基因c DNA全长序列,并应用荧光定量RT-PCR技术测定该基因m RNA的相对表达量,探讨该基因在不同规格四川裂腹鱼鳃、脑、心、肝、脾、肾、肌肉、精巢、卵巢组织中的表达差异,以及温度对其早期仔鱼阶段该基因表达的影响。四川裂腹鱼Cyp19b基因的c DNA全长序列共3 021 bp,共编码507个氨基酸,属于脑型芳香化酶基因;四川裂腹鱼Cyp19b基因编码的氨基酸序列与其他鱼类脑型芳香化酶基因编码的氨基酸序列同源性可达70%以上,而与性腺型芳香化酶基因编码的氨基酸序列同源性较低,为64%左右;四川裂腹鱼Cyp19b基因仅在脑组织中表达,具有比较高的组织特异性,且随着个体的增长,逐渐呈现显著的雌雄差异;对不同温度处理12日龄仔鱼6 d后,低温(10℃和14℃)能显著促进Cyp19b基因表达量的升高,但高温(26℃)却对其表达量无显著影响。由此推测,脑型芳香化酶基因Cyp19b可能在低温导致四川裂腹鱼雌性化过程中发挥着重要作用。     

组织工程面临的技术挑战与发展趋势

当前组织工程研究仍处于初级阶段,还仅仅是初步应用组织工程技术修复临床简单组织缺损,还有许多制约组织工程应用与发展的基本科学问题没有阐明.随着组织工程各个层面技术难题的逐个攻破,组织工程的内涵和外延将不断拓展,并有助于加快组织工程的产业化进程,促进临床应用.针对组织工程核心要素研究的不足,结合最新的组织工程研究进展,阐述...