THE CHARACTERIZATION OF 2 COPPER PHTHALOCYANINE LANGMUIR-BLODGETT-FILMS AND THEIR GAS-SENSITIVE PROPERTIES

The monolayer and deposition behaviour of a symmetrically substituted copper tetra-4-(2, 4-di-t-amylphenoxy) phthalocyanine (tapCuPc) and an asymmetrically substituted copper [tri-4-(2, 4-di-t-amylphenoxy)-mono-4-(-2-methoxyethoxy)]phthalocyanine (AsyCuPc) were investigated. The results on monolayer behaviour and spectroscopic characterization of the LB films show that both CuPc molecules in a monolayer at the air-water interface and the LB films are stacked and inclined. The gas-sensitive properties show that the responding speed of AsyCuPc LB film is faster than that of tapCuPc LB film.

A new naphthoquinoneimine derivative from the marine algal-derived endophytic fungus Aspergillus niger EN-13

Cultivation of an endophytic fungus Aspergillus niger EN-13 that was isolated from the inner tissue of the marine brown alga Colpomenia sinuosa resulted in the characterization of a new naphthoquinoneimine derivative, namely, 5,7-dihydroxy-2-[1-(4methoxy-6-oxo-6H-pyran-2-yl)-2-phenylethylaniino]-[1,4]naphthoquinone. The structure of the new compound was established on the basis of various NMR spectroscopic analyses including 2D NMR techniques, EI-MS, and HR-ESI-MS. This compound displayed moderate antifungal activity. (c) 2007 Bin Gui Wang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

四株海洋放线菌次级代谢产物的研究

本课题组自1999年以来，将培养条件优化、生物活性跟踪及化学跟踪技术应用到胶州湾海洋放线菌的次级代谢产物研究中，发现了一批具有生物活性的化合物，包括新型骨架等新颖结构的化合物。 本研究从海州湾分离出海洋放线菌172株，对其中70株菌的次级代谢产物进行了生物活性筛选和化学筛选，获得了它们对八种病原微生物的抑制活性数据。发现海州湾海洋放线菌对至少一种受试微生物具有拮抗能力的比例约为30%。从海洲湾分离到的海洋放线菌中筛选得到三株L083、L078和L158用于次级代谢产物的研究，同时本人又从合作实验室获得另一株海洋放线菌B7651，从这四株海洋细菌的发酵粗提物中共分离纯化得到26 个化合物， 其中10 个为新颖结构化合物。具体是，3-Hydroxy-6-[(Z)-3´-hydroxy-2´,4´-dimethyl-hept-4´-enoylamino]-2,4-dimethyl-5-oxo-hexanoic acid (4)，2-[5-(2-oxopropyl)tetrahydrofuran-2-yl]propanoic acid (5)，2-oxatricyclo-octane (10)，Huaiomycin (15)， 5-(6-hydroxy-6-methylheptyl)dihydrofuran-2(3H)-one (17), 6-Hydroxy-6-hydroxymethyl-6H-pyran-3-one (18), 1,6-dihydroxy-hex-3-ene-2,5—dione (19) (1’R, 2R, 4R)-2-(1-hydroxy-8-methylnonyl)-4-hydroxymethyl-butanolide (20) ， Bremeromycin A (22) ，Bremeromycin B (26)。生物活性实验结果表明Bremeromycin A (22)具有选择性的抗枯草杆菌(Bacillus subtilis ATCC 6051)活性和抗微藻Chlorella vulgaris活性。

迟缓爱德华氏菌(Edwardsiella tarda) LuxS/AI-2群体感应系统研究

从患病牙鲆中分离出迟缓爱德华氏菌株TX1，经报告菌株检测发现TX1有AI-2活性。用梯度PCR和Genome walking的方法克隆了TX1 luxS基因，将luxS基因在大肠杆菌DH5α中表达，证明其具有功能活性。在TX1中，luxS的表达与AI-2的活性基本是一致的，二者均受生长时期和生长条件的调节，即在glucose存在的条件下luxS表达和AI-2活性升高，而在高温条件下luxS表达和AI-2活性降低。glucose对AI-2活性以及luxS表达的影响经过荧光定量PCR，启动子活性检测，AI-2活性检测以及凝胶滞缓等一系列的实验证实是由cAMP-CRP复合物介导的，该复合物可以通过与luxS启动子相互作用而抑制luxS的表达。RNA干扰表明，TX1中luxS表达被干扰以后，对细菌产生了多方面的影响，包括：(1) 降低AI-2水平；(2) 降低细菌的生长能力；(3) 降低Ⅲ型分泌系统相关基因的表达水平以及生物膜的形成能力；(4) 减弱细菌毒力。外源AI-2的添加可以回复Ⅲ型分泌系统相关基因的表达水平以及生物膜的形成，但是并不能修复生长状况，表明LuxS在TX1中具有双重功能，即参与细胞代谢以及群体感应信号传导。基于LuxS/AI-2群体感应系统对细菌毒力的重要性，设计并筛选了一个该系统的阻遏因子5411。Pull-down实验证明5411可以和LuxS特异性结合。研究表明5411在TX1中表达导致细菌毒力显著下降。将5411克隆至牙鲆共生菌FP3中，发现5411可以被分泌到胞外并能被TX1吸收。将表达5411的共生菌导入牙鲆，发现其能够有效阻遏TX1对牙鲆的侵染。 这些结果表明：(1) TX1中AI-2的活性受控于LuxS，而后者则受生长时期和生长条件的调控；(2) luxS的正常表达对于细菌的正常生长和侵染是必需的；(3) LuxS/AI-2群体感应系统调控Ⅲ型分泌系统相关毒力因子的表达；(4) 通过阻遏LuxS/AI-2群体感应系统来抑制病原菌侵染是一种具有潜力的新型病害防控方法。

4-6岁儿童对情绪表达规则的认知发展研究

情绪表达规则是指一个人应当在恰当的情境中表达恰当的情绪。对情绪表达规则的认知发展是个体社会能力发展的一个重要方面，因此这一主题引起了国内外众多研究者的重视。但目前对童年早期儿童的研究中，大多数只考查了儿童对情绪的外显表达与内隐真实体验的区分，没有同时考查儿童对需要使用情绪表达规则的情境的认知，不能全面真实地揭示儿童对情绪表达规则的认知发展。在本研究中，以个别访谈法考查了4－6岁儿童同一个体在这两个指标上对情绪表达规则认知的发展，以及发展中的个别差异和个体内部差异，并进一步了解情境因素以及心理理论能力、执行抑制能力的发展对它们的影响。主要结果如下： （1）4－6岁儿童对情绪表达规则的认知能力随年龄增长而提高：4岁尚处于发展的萌芽水平，5岁和6岁都处于发展的过渡水平。对于同一年龄的儿童，对情绪表达规则的认知发展存在较大的个体之间的差异。 （2）4－6岁儿童对情绪表达规则的认知也存在较大的个体内部差异：对表情伪装的认知滞后于对需要使用情绪表达规则的情境的认知，对言语伪装的认知优于对表情伪装的认知；在对情绪表达规则的使用中，自我定向的占多数，其次是他人定向，规则定向所占的比例最少。随年龄的增长，自我定向的比例减少，他人定向的比例增多。 （3）情境变量影响4－6岁儿童对情绪表达规则的认知：与同伴交往情境中对需要使用情绪表达规则的情境的认知、对言语伪装的认知优于与长辈交往情境中的有关认知；对需要掩藏消极情绪情境中对表情伪装的认知要优于对需要掩藏积极情绪情境中表情伪装的认知。 （4）4－6岁儿童心理理论能力的发展和执行抑制能力的发展与其对情绪表达规则的认知发展有关。 （5）有关情绪表达规则的提示能够促进儿童对情绪隐私的认知，促进的效果随年龄的增长而提高，6岁的效果最好。

Analysis of phenols by high-performance liquid chromatography with pre-column derivatization by 2-(9-carbazole)-ethyl-chloroformate

2-(9-Carbazole)-ethyl-chloroformate (CEOC), a novel pre-column fluorescence labeling reagent, has been synthesized and applied for the derivatization of phenols. Taken phenol, p-chlorophenol, 2,5-dimethylphenol, 2,4-dichlorophenol and 1,4-dihydroxybenzene as testing standards, the effects of derivatization conditions, such as pH of borate buffer, reaction time and fluorescent tagging reagent concentration, have been systematically studied. Under the optimized conditions, CEOC reacts readily with the phenols to form stable derivatives with excitation and emission wavelengths, respectively, at 293 and 360 nm. The single step derivatization reaction could be finished within 20 min even at room temperature. Such a method has been successfully applied to the analysis of phenols in printing ink by high-performance liquid chromatography. (c) 2005 Elsevier B.V. All rights reserved.

Sleep Patterns in Elementary School Children (Grades 2-5) and Adolescents (Grade 10)

Background: To date, there is limited research examining sleep patterns in elementary school children. Previous researchers focused on parental responses rather than student responses to determine factors that affect sleep. The presented study surveyed sleep patterns and examined external factors affecting total sleep time among elementary school children and adolescents. Methods: Students in grades 2-5 (n=885) and grade 10 (n=190) enrolled in a public school system in the Northeast, completed a district administered survey that included questions on sleep duration and hygiene. Results. Average reported sleep duration decreased with increasing grade level. Children in grades 2-5 woke up earlier (31.7-72.4%) and on their own in comparison to adolescents in grade 10 (6.8%). Significantly shorter sleep durations were associated with having a television (grades 2, 4, 5, p< 0.01) or a cell phone in the room (grades 3, 4; p < 0.05), playing on the computer or video games (grades 3, 4, p<.001) before going to bed. In contrast, students in grade 2, 3, & 4 who reported reading a book before going to bed slept on average 21 minutes more per night (p=.029, .007, .009, respectively). For tenth graders, only consumption of energy drinks led to significant reduction in sleep duration (p<.0001). Conclusion. Sleep is a fundamental aspect in maintaining a healthy and adequate life style. Understanding sleep patterns will assist parents, health care providers, and educators in promoting quality sleep hygiene in school-aged children.

Physiological levels of PTEN control the size of the cellular Ins(1,3,4,5,6)P5 pool

To understand how a signaling molecule's activities are regulated, we need insight into the processes controlling the dynamic balance between its synthesis and degradation. For the Ins(1,3,4,5,6)P5 signal, this information is woefully inadequate. For example, the only known cytosolic enzyme with the capacity to degrade Ins(1,3,4,5,6)P5 is the tumour-suppressor PTEN [J.J. Caffrey, T. Darden, M.R. Wenk, S.B. Shears, FEBS Lett. 499 (2001) 6 ], but the biological relevance has been questioned by others [E.A. Orchiston, D. Bennett, N.R. Leslie, R.G. Clarke, L. Winward, C.P. Downes, S.T. Safrany, J. Biol. Chem. 279 (2004) 1116 ]. The current study emphasizes the role of physiological levels of PTEN in Ins(1,3,4,5,6)P5 homeostasis. We employed two cell models. First, we used a human U87MG glioblastoma PTEN-null cell line that hosts an ecdysone-inducible PTEN expression system. Second, the human H1299 bronchial cell line, in which PTEN is hypomorphic due to promoter methylation, has been stably transfected with physiologically relevant levels of PTEN. In both models, a novel consequence of PTEN expression was to increase Ins(1,3,4,5,6)P5 pool size by 30-40% (p<0.01); this response was wortmannin-insensitive and, therefore, independent of the PtdIns 3-kinase pathway. In U87MG cells, induction of the G129R catalytically inactive PTEN mutant did not affect Ins(1,3,4,5,6)P(5) levels. PTEN induction did not alter the expression of enzymes participating in Ins(1,3,4,5,6)P5 synthesis. Another effect of PTEN expression in U87MG cells was to decrease InsP6 levels by 13% (p<0.02). The InsP6-phosphatase, MIPP, may be responsible for the latter effect; we show that recombinant human MIPP dephosphorylates InsP6 to D/L-Ins(1,2,4,5,6)P5, levels of which increased 60% (p<0.05) following PTEN expression in U87MG cells. Overall, our data add higher inositol phosphates to the list of important cellular regulators [Y. Huang, R.P. Wernyj, D.D. Norton, P. Precht, M.C. Seminario, R.L. Wange, Oncogene, 24 (2005) 3819 ] the levels of which are modulated by expression of the highly pleiotropic PTEN protein.

Total Synthesis of (±)-Phomactin G, a Platelet Activating Factor Antagonist from the Marine Fungus Phoma sp.

A total synthesis of phomactin G (3), which is a central intermediate in the biosynthesis of phomactin A (5) in Phoma sp. is described. The synthesis is based on a Cr(II)/Ni(II) macrocyclisation from the aldehyde vinyl iodide 9, leading to 16, followed by sequential conversion of 16 into the -epoxide 21 and the ketone 25 which, on deprotection, led to (±)-phomactin G. Phomactin G (3) shares an interesting structural homology with phomactin D (2), the most potent PAF-antagonist metabolite in Phoma sp. It is most likely converted into phomactin A (5), by initial allylic oxidation to the transient -alcohol phomactin structure 4, known as Sch 49028, followed by spontaneous pyran ring formation.

New mustard-linked 2-aryl-bis-benzimidazoles with anti-proliferative activity

We describe new methodology for the synthesis of symmetric bis-benzimidazole carrying 2-aryl moieties, including 2-[4-3'-aminopropoxy)phenyl] and 2-[4-(3'-aminopropanamido)pheny] substituents, together with the synthesis of novel hybrid molecules comprising bis-benzimidazoles in ester and amide combination with the N-mustard chlorambucil. The in vitro activities of these compounds against five cancer cell lines are also provided.

Down-regulation of beta1,4-galactosyltransferase V is a critical part of etoposide-induced apoptotic process and could be mediated by decreasing Sp1 levels in human glioma cells.

beta1,4-Galactosyltransferase V (beta1,4GalT V; EC 2.4.1.38) is considered to be very important in glioma for expressing transformation-related highly branched N-glycans. Recently, we have characterized beta1,4GalT V as a positive growth regulator in several glioma cell lines. However, the role of beta1,4GalT V in glioma therapy has not been clearly reported. In this study, interfering with the expression of beta1,4GalT V by its antisense cDNA in SHG44 human glioma cells markedly promoted apoptosis induced by etoposide and the activation of caspases as well as processing of Bid and expression of Bax and Bak. Conversely, the ectopic expression of beta1,4GalT V attenuated the apoptotic effect of etoposide on SHG44 cells. In addition, both the beta1,4GalT V transcription and the binding of total or membrane glycoprotein with Ricinus communis agglutinin-I (RCA-I) were partially reduced in etoposide-treated SHG44 cells, correlated well with a decreased level of Sp1 that has been identified as an activator of beta1,4GalT V transcription. Collectively, our results suggest that the down-regulation of beta1,4GalT V expression plays an important role in etoposide-induced apoptosis and could be mediated by a decreasing level of Sp1 in SHG44 cells, indicating that inhibitors of beta1,4GalT V may enhance the therapeutic efficiency of etoposide for malignant glioma.

Beta1,4-galactosyltransferase V functions as a positive growth regulator in glioma.

beta1,4-galactosyltransferase V (GalT V; EC 2.4.1.38) can effectively galactosylate the GlcNAcbeta1-->6Man arm of the highly branched N-glycans that are characteristic of glioma. Previously, we have reported that the expression of GalT V is increased in the process of glioma. However, currently little is known about the role of GalT V in this process. In this study, the ectopic expression of GalT V could promote the invasion and survival of glioma cells and transformed astrocytes. Furthermore, decreasing the expression of GalT V in glioma cells promoted apoptosis, inhibited the invasion and migration and the ability of tumor formation in vivo, and reduced the activation of AKT. In addition, the activity of GalT V promoter could be induced by epidermal growth factor, dominant active Ras, ERK1, JNK1, and constitutively active AKT. Taken together, our results suggest that GalT V functioned as a novel glioma growth activator and might represent a novel target in glioma therapy.

Elevated beta1,4-galactosyltransferase I in highly metastatic human lung cancer cells. Identification of E1AF as important transcription activator.

The elevated levels of beta1,4-galactosyltransferase I (GalT I; EC 2.4.1.38) are detected in highly metastatic lung cancer PGBE1 cells compared with its less metastatic partner PGLH7 cells. Decreasing the GalT I surface expression by small interfering RNA or interfering with the surface of GalT I function by mutation inhibited cell adhesion on laminin, the invasive potential in vitro, and tyrosine phosphorylation of focal adhesion kinase. The mechanism by which GalT I activity is up-regulated in highly metastatic cells remains unclear. To investigate the regulation of GalT I expression, we cloned the 5'-region flanking the transcription start point of the GalT I gene (-1653 to +52). Cotransfection of the GalT I promoter/luciferase reporter and the Ets family protein E1AF expression plasmid increased the luciferase reporter activity in a dose-dependent manner. By deletion and mutation analyses, we identified an Ets-binding site between nucleotides -205 and -200 in the GalT I promoter that was critical for responsiveness to E1AF. It was identified that E1AF could bind to and activate the GalT I promoter by electrophoretic mobility shift assay in PGLH7 cells and COS1 cells. A stronger affinity of E1AF for DNA has contributed to the elevated expression of GalT I in PGBE1 cells. Stable transfection of the E1AF expression plasmid resulted in increased GalT I expression in PGLH7 cells, and stable transfectants migrated faster than control cells. Meanwhile, the content of the beta1,4-Gal branch on the cell surface was increased in stably transfected PGLH7 cells. GalT I expression can also be induced by epidermal growth factor and dominant active Ras, JNK1, and ERK1. These data suggest an essential role for E1AF in the activation of the human GalT I gene in highly metastatic lung cancer cells.

Demonstration of increased concentrations of circulating glycated insulin in human Type 2 diabetes using a novel and specific radioimmunoassay.

Aims/hypothesis: Glycation of insulin, resulting in impaired bioactivity, has been shown within pancreatic beta cells. We have used a novel and specific radioimmunoassay to detect glycated insulin in plasma of Type 2 diabetic subjects.

Methods: Blood samples were collected from 102 Type 2 diabetic patients in three main categories: those with good glycaemic control with a HbA1c less than 7%, moderate glycaemic control (HbA1c 7–9%) and poor glycaemic control (HBA1c greater than 9%). We used 75 age- and sex-matched non-diabetic subjects as controls. Samples were analysed for HbA1c, glucose and plasma concentrations of glycated insulin and insulin.

Results: Glycated insulin was readily detected in control and Type 2 diabetic subjects. The mean circulating concentration of glycated insulin in control subjects was 12.6±0.9 pmol/l (n=75). Glycated insulin in the good, moderate and poorly controlled diabetic groups was increased 2.4-fold (p<0.001, n=44), 2.2- fold (p<0.001, n=41) and 1.1-fold (n=17) corresponding to 29.8±5.4, 27.3±5.7 and 13.5±2.9 pmol/l, respectively.

Conclusion/interpretation: Glycated insulin circulates at noticeably increased concentrations in Type 2 diabetic subjects. [Diabetologia (2003) 46:475–478]