Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin

Defensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.

Identification of a novel cathelicidin gene in the rainbow trout, Oncorhynchus mykiss

We report the cloning of a novel antimicrobial peptide gene, termed rtCATH_1, found in the rainbow trout, Oncorhynchus mykiss. The predicted 216-residue rtCATH_1 prepropeptide consists of three domains: a 22-residue signal peptide, a 128-residue cathelin-like region containing two identifiable cathelicidin family signatures, and a predicted 66-residue C-terminal cationic antimicrobial peptide. This predicted mature peptide was unique in possessing features of different known (mammalian) cathelicidin subgroups, such as the cysteine-bridged family and the specific amino-acid-rich family. The rtCATH_1 gene comprises four exons, as seen in all known mammalian cathelicidin genes, and several transcription factor binding sites known to be of relevance to host defenses were identified in the 5' flanking region. By Northern blot analysis, the expression of rtCATH_1 was detected in gill, head kidney, and spleen of bacterially challenged fish. Primary cultures of head kidney leukocytes from rainbow trout stimulated with lipopolysaccharide or poly(I (.) C) also expressed riCATH_1. A 36-residue peptide corresponding to the core part of the fish cathelicidin was chemically synthesized and shown to exhibit potent antimicrobial activity and a low hemolytic effect. Thus, rtCATH_1 represents a novel antimicrobial peptide gene belonging to the cathelicidin family and may play an important role in the innate immunity of rainbow trout.

Water solubility enhancement of phthalates by cetyltrimethylammonium bromide and beta-cyclodextrin

Water solubility enhancements of six phthalates (five aliphatic phthalates and one phenyl phthalate) by cetyltrimethylammonium bromide (CTAB) and beta-cyclodextrin (beta-CD) were studied at 25 degreesC. The solubilities of these plithalates are remarkably enhanced by CTAB solutions above the critical micelle concentration (cmc). Only marginal enhancement of phthalate solubility was observed in solutions containing CTAB below its cmc and beta-CD at low concentrations (less than 5 mM). The solubility enhancements of the plithalates are proportional to the added amount of CTAB and beta-CD. Partition coefficients of the plithalates between monomeric CTAB surfactant and water (K-MN) and between CTAB micelle and water K-MC) were estimated from the experimental data. The mechanisms of solubility enhancements by CTAB and beta-CD were discussed. A log-linear equation was proposed and evaluated for the solubilization by CTAB below cmc, while the previously proposed linear partitioning model was questioned. The structures of the complexes formed between plithalates and beta-CD were proposed, and the formation constants were estimated. The values of log K-MC, log K-MN, and log Kbeta-CD of the plithalates were found to correlate linearly with the log K-OW of plithalates, with the exception of the solid phenyl phthalate.

Polymerization of ionic liquid-based microemulsions: A versatile method for the synthesis of polymer electrolytes

电子邮箱fyan@suda.edu.cn

气体水合物生成特性实验研究

实验研究了添加剂对甲烷气体水合物形成过程的影响。发现微量的表面活性剂降低了甲烷气体水合物在静止反应器中形成的诱导时间，并使水合物快速形成和生长，提高了水合物形成过程中的填充密度。阴离子表面活性剂（十二烷基硫酸钠）对水合物生长的促进作用比非离子表面活性剂（烷基多糖苷）强。液态烃环戊烷降低了水合物形成的诱导时间，但环戊烷不能提高水合物的填充密度。

Study on methane hydration process in a semi-continuous stirred tank reactor

The methane hydration process is investigated in a semi-continuous stirred tank reactor. Liquid temperatures and reaction rates without stirrer are compared with those occurring with stirrer, while at the same time better stirring conditions of the methane hydration process are given by the experiments. Some basic data of fluid mechanics, for example, stirring Reynolds number, Froucle number and stirrer power, are calculated during the methane hydration process, which can be applied to evaluate stirrer capacity and provide some basic data for a scaled up reactor. Based on experiment and calculations in this work, some conclusions are drawn. First, the stirrer has great influence on the methane hydration process. Batch stirring is helpful to improve the mass transfer and heat transfer performances of the methane hydration process. Second, induction time can be shortened effectively by use of the stirrer. Third, in this paper, the appropriate stirring velocity and stirring time were 320 rpm and 30 min, respectively, at 5.0 MPa, for which the storage capacity and reaction time were 159.1 V/V and 370 min, respectively. Under the condition of the on-flow state, the initial stirring Reynolds number of the fluid and the stirring power were 12,150 and 0.54 W, respectively. Fourth, some suggestions, for example, the use of another type of stirrer or some baffles, are proposed to accelerate the methane hydration process. Comparing with literature data, higher storage capacity and hydration rate are achieved in this work. Moreover, some fluid mechanics parameters are calculated, which can provide some references to engineering application.

插层剂在聚合物熔融插层蒙脱土中的作用

近年来，由于有机和无机成分在纳米尺寸上的结合而带来的优异性能使得聚合物／蒙脱土纳米复合材料得到广泛关注。在诸多方法中，因能够使用传统的聚合物熔融挤出、共混等加工设备而无需额外的设备投资，投资低、见效快而且对周围环境没有污染等优点，熔融插层法倍受青睐。然而，通常认为亲油性聚合物无法直接插层蒙脱土，而需要预先用插层剂处理蒙脱土。大多数聚合物是亲油性的，这将大大增加制备材料的成本和工艺的复杂性，并因引入了小分子插层剂而有损材料性能的提高，从而阻碍了该技术的应用和推广。本文从理论上研究了插层剂用量对有机化蒙脱土及聚合物／蒙脱土纳米复合材料微观结构的影响，探索一种无需改性蒙脱土或用很少插层剂改性蒙脱土的制备聚合物／蒙脱土纳米复合材料的方法。1．系统地改变插层剂用量，制备不同的有机化蒙脱土。通过广角X射线衍射、热失重分析、索氏抽提器抽提、差热分析和付立叶红外分析等表征手段，研究了插层剂插层机理以及插层剂在蒙脱土层间的排列方式，为进一步研究有机化蒙脱土在聚合物树脂中的分散奠定基础。结果表明，插层剂插层蒙脱土机理和在蒙脱土层间的排列方式随着插层剂用量的变化而不同。2．用不同的亲油性聚合物熔融插层有机化蒙脱土以制备相应的聚合物／蒙脱土纳米复合材料。用广角嘴射线衍射仪和透射电子显微镜分析研究了蒙脱土在聚合物／蒙脱土纳米复合材料中分散状态。结果表明，随着插层剂用量的减少，蒙脱土片层在聚合物树脂中的分散逐渐由插层型过渡到剥离型，即减少改性蒙脱土时插层剂用量有利于蒙脱土在纳米尺寸上分散于聚合物树脂中。3．用不同的亲油性聚合物直接熔融插层未改性的蒙脱土，广角X射线衍射仪和透射电子显微镜表征结果表明，与蒙脱土片层有相互作用的亲油性聚合物能够直接熔融插层未改性的蒙脱土，获得剥离或插层型聚合物／蒙脱土纳米复合材料，这对于蒙脱土在聚合物改性中的应用具有重要的实际意义和理论意义。4．用共同的母料制备聚丙烯／酸配化聚丙烯／蒙脱土纳米复合材料和尼龙12／酸醉化聚丙烯／蒙脱土纳米复合材料，用广角X射线衍射仪和透射电子显微镜对比分析蒙脱土在聚合物中分散的差异，从而了解聚丙烯与酸配化聚丙烯之间的增容作用对蒙脱土在聚丙烯树脂中分散的影响；同时，用尼龙11和酸配化聚丙烯共插层蒙脱土制备聚丙烯／蒙脱土纳米复合材料，并探讨其增容机理。

表面活性剂与菲的复合生态效应

利用表面活性剂去除土壤中难降解的多环芳烃，近年来得到了较多的应用.但对其去污机理及两者共存时的复合毒性，国内外研究甚少.该篇论文以六种不同离子类型、分子结构的表面活性剂和以菲为多环芳烃的代表，研究了表面活性剂对菲降解的影响，和与菲共存时的复合毒性，以及表面活性剂对菲在模拟生态系统中的迁移转化规律的影响.

三孢布拉氏霉发酵产β-胡萝卜素的研究

本论文研究了利用三孢布拉氏霉(Blakeslea trispora)发酵产β-胡萝卜素的培养条件。主要包括：发酵培养基的确定，发酵条件的优化。还考察了发酵菌丝体中β-胡萝卜素的提取方法及薄层层析等。 首先研究了培养基成分对三孢布拉氏霉发酵产β-胡萝卜素的影响。确立了玉米淀粉作为碳源，黄豆粉（热榨）作为氮源，棉籽油作为植物油的发酵培养基配方，其成分为：玉米淀粉 3％，黄豆粉（热榨） 2％，棉籽油 3％，KH2PO4 0.2%，MgSO4·7H2O 0.2%，维生素B1 0.002％，pH值6.0。 其次，通过比较不同的发酵影响因子，分别得到最适的条件：如三孢布拉氏霉正负菌接种比例为1.3：0.7，培养基pH值为7.0（灭菌后），发酵促进因子为Triton X-100。并采用正交试验法，确定其最佳发酵条件为正负菌接种比例1.3/0.7，发酵培养基pH为7.0，在培养基中添加表面活性基Triton X-100 0.08％。使该菌株产β-胡萝卜素的量达到0.73g/L，较初始发酵条件提高了3.3倍。 研究中还找到一个简便有效的对β-胡萝卜素的提取方法，选用盐酸-热处理法进行细胞破壁，并选用沸程为60～90℃的石油醚进行萃取。 用三孢布拉霉菌丝体内类胡萝卜索的石油醚提取液点样于硅胶G板，以丙酮：石油醚（5：95）为展开剂能将β-胡萝卜素与其它类胡萝卜索分离。该方法简便快速，并有一定实用价值。 The fermentative conditions of β-carotene by Blakeslea trispora have been investigated. These conditions include fermentation medium, the optimization of some fermentation factor. The extracting methods and the TLC of carotenoids were also researched. Firstly, the effects of composition of fermentation medium on the yield of β-carotene were studied. the results showed that the best fermentation medium was corn starch 3％，soybean power 2％，cottonseed oil 3％，KH2PO4 0.2%，MgSO4·7H2O 0.2%，vitamin B1 0.002％，pH value 6.0. Secondly, through compared some factors, such as different proportion of plus and minus strains, pH value, nonionic surfactants, respective best values have been obtained. The best proportion of plus and minus strains is 1.3:0.7, pH value of fermentation medium (sterilized) is 7.0, fermentation accelerant which acts as surfactants is Triton x-100. Farther on, the fermentative conditions were optimized through orthogonal experiment, the optimization showed that proportion of plus and minus strains is 1.3:0.7，pH value is 7.0, content of Triton x-100 is 0.08％. And the yield of β-carotene reached 0.73g/L, which was up to 3.3 times through the fermentation. In the extracting study, it has showed hydrochloric acid-heat treatment is a simple, convenient and effective extracting methods is which was used to destroy the cell wall, and the extracting organic solvent is petroleum ether whose boiling range is 60~90 ℃. In the TLC experiments, extracting contents in the petroleum ether were spotted in the silicagel plate, and the mixed liquor of acetone and petroleum ether (5：95) is developping agent, which can distinguish β-carotene from other carotenoids. It is a simple and quick technique.