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.     

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。     

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.     

四川市售白芍饮片质量调研

目的:调研四川市售白芍饮片质量。方法:随机采购四川市售白芍饮片16批,参照现行2010版药典的规定,测定饮片水分、总灰分、水溶性浸出物,并采用HPLC法测定芍药苷含量。结果:16批次白芍饮片中仅3批次芍药苷的量符合药典规定,9批次水分符合药典规定。结论:白芍饮片存在的质量问题主要是芍药苷含量和水分含量不合格。针对导致白芍药材或饮片质量问题的多种原因,药品监督管理部门应加强监督管理,规范饮片市场。     

Inhibition of the growth and development of mosquito larvae of Culex quinquefasciatus(Diptera:Culicidae) treated with extract from leaves of Pseudocalymma alliaceum(Bignonaceae)

Objective:To determine larvicidal activity of the essential oil,hydrolat and botanical extracts derived from leaves ol Pseudocalymma alliaceum an mosquito larvae of Culex quinquefasciatus.Methods:Groups of twenty lanae were used in the larvicidal assays.The mortality,relative growth rate,the larval and pupal duration and viability was estimated.The essential oil was analyzed by solid phase microextraction using gas chromatography coupled to mass spectrometry.Results:Essential oil at 800 ppm showed larvicidal activity at 24 h with lethal values of LC_(50) and LC_(90) of 267.33 and 493.63 ppm.The hvdrolat at 20% and 10% on 2nd stage larvae showed 100%effectiveness after 24 h.The aqueous extract at 10% had a relative growth index of 0.58.while the ethanolic and methanolic extract obtained values of 0.76 and 0.70 and control reached 0.99.Larvae treated with 10% of methanol,ethanol and aqueous extract showed a reduction in larval duration of 5.00,2.20 and 4.35 days;ethanol extract at 1% provoke decrease of 2.40 days in the development and exhibited an increment of 3.30 days when treated with 0.01%.Aqueous,ethanol and methanol extracts at 10%reduced in 6.15,3.42 and 5.57 days pupal development.The main compounds were diallyl disulfide(50.05%),diallyl sulfide(11.77%) and disulfide di-2-propenyl(10.37%).Conclusions:The study demonstrated for the first time,the larvicidal activity of the essential oil and hydrolat of Pseudocalymma alliaceum:aqueous,ethanol and methanol extracts inhibited the normal growth and development of mosquito larvae,prolonging and delaying larval and pupal duration.