Comparative analysis of protein profiles of aqueous extracts from marine sponges and assessment of cytotoxicity on different mammalian cell types

Marine natural products extracted from sponges represent a new source for drug discovery. Here we describe a simple method for preparing aqueous extracts from 7 Mediterranean demosponges, which allowed the extraction of water-soluble compounds, such as proteins by homogenization of sponge tissue in phosphate buffered saline (PBS).The comparative analysis by SDS-PAGE showed differences in number of bands, bandwidth and intensity among the sponges analyzed. The PAS/silver staining revealed a substantial and different glycoprotein assortment among the demosponges studied.To further study the biological activities present in the sponge extracts, we determined the non-cytotoxic doses on four different mammalian cell types demonstrating that the optimal non-cytotoxic doses were cell type- and extract-dependent.In conclusion, the extraction method described in this paper represents a fast and efficient procedure for the extraction of water-soluble proteins from marine sponges. Furthermore, the cell viability data suggest the feasibility of this method for the direct in vitro cell-based assays.     

Induction and treatment of anergy in murine leprosy

Leprosy is a disease consisting of a spectrum of clinical, bacteriological, histopathological and immunological manifestations. Tuberculoid leprosy is frequently recognized as the benign polar form of the disease, while lepromatous leprosy is regarded as the malignant form. The different forms of leprosy depend on the genetic and immunological characteristics of the patient and on the characteristics of the leprosy bacillus. The malignant manifestations of lepromatous leprosy result from the mycobacterial‐specific anergy that develops in this form of the disease. Using murine leprosy as a model of anergy in this study, we first induced the development of anergy to Mycobacterium lepraemurium (MLM) in mice and then attempted to reverse it by the administration of dialysable leucocyte extracts (DLE) prepared from healthy (HLT), BCG‐inoculated and MLM‐inoculated mice. Mice inoculated with either MLM or BCG developed a robust cell‐mediated immune response (CMI) that was temporary in the MLM‐inoculated group and long‐lasting in the BCG‐inoculated group. DLE were prepared from the spleens of MLM‐ and BCG‐inoculated mice at the peak of CMI. Independent MLM intradermally‐inoculated groups were treated every other day with HLT‐DLE, BCG‐DLE or MLM‐DLE, and the effect was documented for 98 days. DLE administered at a dose of 1.0 U (1 × 10⁶ splenocytes) did not affect the evolution of leprosy, while DLE given at a dose of 0.1 U showed beneficial effects regardless of the DLE source. The dose but not the specificity of DLE was the determining factor for reversing anergy.     

厚朴乳膏处方筛选及制备工艺的研究与开发

目的厚朴酚性成分具有杀菌、消炎、燥湿、止痒等作用,通过制备乳膏对其提取物进行开发利用。方法根据厚朴酚、和厚朴酚的特点及乳膏基质的特点,筛选处方组成,并对制备工艺进行研究,通过对乳膏的外观性状、透皮、稠度和稳定性等指标进行考察,筛选最佳的处方组成,并制备厚朴乳膏。结果由该比例的组成制备的厚朴乳膏,重现性好,乳膏色泽较纯正,均匀细腻,稠度适宜,易涂布,稳定性好、透皮吸收等均较好,可以作为最佳配比来制备厚朴乳膏。结论厚朴乳膏可以作为一种新制剂进行开发利用。     

Ovicidal and Oviposition Deterrent Activities of Medicinal Plant Extracts Against Aedes aegypti L. and Culex quinquefasciatus Say Mosquitoes (Diptera: Culicidae)

Objectives

To evaluate the ovicidal and oviposition deterrent activities of five medicinal plant extracts namely Aegle marmelos (Linn.), Limonia acidissima (Linn.), Sphaeranthus indicus (Linn.), Sphaeranthus amaranthoides (burm.f), and Chromolaena odorata (Linn.) against Culex quinquefasciatus and Aedes aegypti mosquitoes. Three solvents, namely hexane, ethyl acetate, and methanol, were used for the preparation of extracts from each plant.

Methods

Four different concentrations—62.5 parts per million (ppm), 125 ppm, 250 ppm, and 500 ppm—were prepared using acetone and tested for ovicidal and oviposition deterrent activities. One-way analysis of variance (ANOVA) was used to determine the significance of the treatments and means were separated by Tukey''s test of comparison.

Results

Among the different extracts of the five plants screened, the hexane extract of L. acidissima recorded the highest ovicidal activity of 79.2% and 60% at 500 ppm concentration against the eggs of Cx. quinquefasciatus and Ae. aegypti, respectively. Similarly, the same hexane extract of L. acidissima showed 100% oviposition deterrent activity at all the tested concentrations against Cx. quinquefasciatus and Ae. aegypti adult females.

Conclusion

It is concluded that the hexane extract of L. acidissima could be used in an integrated mosquito management program.     

皱瘤海鞘乙醇提取物对HSV-2基因复制的影响

目的：研究皱瘤海鞘乙醇提取物对HSV-2 DNA复制的影响;方法：以Vero细胞为模型,应用CPE法,MTT法、real-time PCR与反向PCR检测皱瘤海鞘乙醇提取物对HSV-2DNA合成的抑制作用。结果：皱瘤海鞘乙醇提取物显著抑制HSV-2所致的CPE;皱瘤海鞘乙醇提取物在HSV-2生物合成早期(-2,0,2小时)时干预,对HSV-2的生物合成抑制效果较好,病毒抑制率达65%以上;皱瘤海鞘乙醇提取物能够显著抑制HSV-2 DNA拷贝数,与病毒对照组比较具有显著差异(P＜0.01),并能下调HSV-2DNA聚合酶UL30基因的表达。结论：皱瘤海鞘对HSV-2DNA复制具有抑制作用,其作用靶点之一是HSV-2DNA聚合酶UL30基因。     

紫草萘醌色素微乳液抑制肝癌细胞增殖的实验研究

目的观察紫草萘醌醇提物对肝癌细胞体内外增殖的影响。方法紫草萘醌醇提取物制备成微乳液(LE);加入人肝癌细胞系、人胚肾细胞系和小鼠成纤维细胞系的体外培养,MTT法检测细胞增殖;小鼠皮下接种肝癌细胞制作荷瘤模型,分别应用LE或5-FU,观察其生活状态、生存时间以及瘤重等指标。结果体外共育48~72h,LE抑制人肝癌细胞系的增殖;这种作用具有浓度依赖特点,而且相同浓度下不影响正常传代细胞的生长。体内应用可明显抑制移植小鼠肝癌的生长,延长荷瘤小鼠生存期;有效剂量2.5~10mg·kg-1·d-1。应用LE还可提高受者小鼠的生活质量。结论无论在体外或体内,LE对肿瘤细胞的增殖都表现量-效依赖的抑制作用。     

植物基除臭剂的配方设计及净化效果的试验研究

目的通过研究植物基除臭剂的配方,探讨其净化效果。方法采用市售植物复配精油为基底,三甘醇为溶剂,加入一定比例的乳化剂、助剂和去离子水配制植物基除臭剂。乳化剂选用比较常见的10种表面活性剂进行冷储和热储对比试验,助剂选用β-环糊精,通过三因素三水平正交试验确定配方中植物精油、乳化剂和助剂含量。通过气相色谱质谱分析除臭剂的活性成分。净化试验采用静态染毒和自动喷雾系统测试除臭剂的净化效果,并和自然沉降、去离子水和3M异味抑制剂效果进行对比。结果植物基除臭剂检出活性有效成分33种,大部分为含氧化合物。精油质量浓度0.6%、β-环糊精质量浓度1.0%,吐温-80质量浓度3.0%为最佳配比,该植物基除臭剂的平均除臭效率高于去离子水和化学除臭剂,10 min后硫化氢和甲硫醇的净化效率达到100%,30 min后氨气净化效率达到95%。结论植物基除臭剂含有反应活性很高的功能团,能与恶臭物质进行各种化学反应,从而达到抑制恶臭气味的目的,该除臭剂对硫化氢和甲硫醇的净化效果强于对氨气的净化效果。     

Cytocompatibility study of NHLBI primary reference materials using human endothelial cells

Four different materials, low density polyethylene (LDPE), polydimethylsiloxane (PDMS), polyvinylcholoride (PVC) and cellulose, were selected by the Devices and Technology Branch of the National Heart Lung and Blood Institute (NHLBI) as primary reference materials for blood contacting. Among the wide variety of tests proposed to assess hemocompatibility of short-term blood contacting catheters, it was desirable to rule out whether these materials could release toxics for vascular cells of the physiological environment. Thus, the cytocompatibility of these materials have been checked towards human umbilical vein endothelial cells: the method used avoids direct contact between cells and materials but evaluates the effect of possible toxic substances leached from materials. These substances were obtained under defined conditions according to a standard. The results show that the extracts of cellulose and LDPE provoke an important cytotoxic effect on the endothelial cell cultures, while the extracts of PDMS and PVC allow the obtention of endothelial cell lining of the reference surface, with a correct global metabolic activity and the intracellular presence of von Willebrand factor.