Evaluation of In Vitro Cytotoxic Property Against Cholangiocarcinoma Cell Line and GC/MS Analysis from Leaf of Erythrophleum succirubrum Gagnep

Objective: Plants are valuable sources of new pharmaceuticals. Secondary metabolites of the genus Erythrophleum exhibit cytotoxicity and may have therapeutic value. The cytotoxic activity of ethanolic leaf extract of Erythrophleum succirubrum Gagnep. against a human cholangiocarcinoma cell line was assessed. Methods: Crude extract of E. succirubrum was prepared by ethanol extraction. The ethanolic leaf extract of E. succirubrum was evaluated for cytotoxicity against the human cholangiocarcinoma cell line KKU-M213 using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assays. The chemical composition of E. succirubrum leaf extract was analyzed using GC/MS. Result: The ethanolic leaf extract of E. succirubrum reduced the viability of KKU-M213 cells in a dose- and time-dependent manner. It showed high cytotoxicity, with IC50 values of 65.22 ± 1.18 µg/mL and 1.19 ± 1.38 µg/mL at exposure times of 24 and 96 h, respectively. GC/MS analysis of the ethanolic leaf extract of E. succirubrum identified 22 components. The main constituents identified were Cyclohexanone, 2-[2-nitro-1-(2-naphthyl)ethyl]-(14.79%) followed by allomycin (14.65%), mome inositol (14.30%), campesterol (11.80%) and ethyl linolenate (10.83%), respectively. Conclusion: Five major groups of compounds were found, with lipids dominating, followed by carbohydrates, benzenoids, phenylpropanoids, polyketides and organoheterocyclic compounds. Many of the bioactive components discovered in the ethanolic leaf extract of E. succirubrum might be responsible for its cytotoxic properties.     

Phytochemical studies on Stemona aphylla: isolation of a new stemofoline alkaloid and six new stemofurans

A new stemofoline alkaloid, (2'S)-hydroxy-(11S,12R)-dihydrostemofoline (3), new stemofurans M-R (8-13), and known compounds stemofoline (1), (2'S)-hydroxystemofoline (2), stemofuran E (4), stemofuran F (5), stemofuran J (6), and stilbostemin F (7) have been isolated from the root extracts of Stemona aphylla. The structures and relative configurations of these new compounds have been determined by spectroscopic data interpretation and from semisynthetic studies. These natural and semisynthetic alkaloids were tested for acetylcholinesterase inhibitory activities and were found to be 10-20 times less active than 1',2'-didehydrostemofoline itself. Stemofurans 4, 6, 8, 11, and 13 were tested for their antibacterial and antifungal activities. Three of these showed antibacterial activities against MRSA with MIC values of 15.6 μg/mL.     

Acute effects of SO2 and particles from a power plant on respiratory symptoms of children, Thailand

Epidemiological studies to evaluate the acute effects of ambient SO2 on the respiratory health of children provide inconclusive results. A panel study to examine the association of short-term exposure to ambient SO2 and respiratory symptoms of 196 children for a period of 107 days was conducted in Thailand. Generalized Estimating Equations were used to examine the association of daily variation of air pollution with daily respiratory symptoms. During the study period, SO2 was not associated with respiratory symptoms in either asthmatics or non-asthmatics, whereas a 10 microg/m3 increase in PM10 was modestly associated with increases of lower respiratory symptom incidence (OR=1.03, 95%CI=0.98, 1.09) and cough (OR=1.04, 95% CI=1.00, 1.08) in asthmatics. At the low ambient air pollution concentrations observed, particulate matter rather than SO2 was associated on a microg/m3 basis with acute daily respiratory symptoms.     

Phytochemical investigations of Stemona curtisii and synthetic studies on stemocurtisine alkaloids

The isolation of two new Stemona alkaloids, 1-hydroxyprotostemonine and stemocurtisine N-oxide, and a new benzofuran, stemofuran L, from the root extracts of Stemona curtisii is reported. The major known alkaloids from this plant extract, stemocurtisine, stemocurtisinol, and oxyprotostemonine, were also isolated along with oxystemokerrine N-oxide. The nonalkaloid components of this extract included a new benzofuran derivative, stemofuran L, the known stemofurans F, J, and K, dihydro-γ-tocopherol, and stigmasterol. Stemocurtisine and stemocurtisinol were converted to their respective N-oxides by oxidation. Stemocurtisine was converted to a tetracyclic derivative by oxidative cleavage of the γ-butyrolactone ring, while stemocurtisinol gave a novel lactam derivative by oxidative cleavage of the C-4 side chain under basic conditions. The acetylcholinesterase inhibitory activities of some known and new alkaloids and their derivatives are also reported. All were 10-20 times less active as acetylcholinesterase inhibitors than the pyrrolo[1,2-a]azepine Stemona alkaloids stemofoline and 1',2'-didehydrostemofoline. None of the stemofuran compounds showed significant antibacterial or antifungal activities.     

New method to produce equine antirabies immunoglobulin F(ab′)2 fragments from crude plasma in high quality and yield

Rabies is still a major cause of human deaths in several developing countries. According to the World Health Organization, administration of antirabies serum or antirabies immunoglobulin is recommended for patients who have experienced a category-III exposure to rabies. Improvement of antirabies immunoglobulin production is required to enhance safety and efficacy of the products. In this paper, a new method to produce equine antirabies immunoglobulin F(ab′)₂ fragments from crude plasma is proposed. First, protein G affinity chromatography was used to purify IgG from equine plasma. Moreover, purification of IgG was shown to facilitate its digestion by pepsin. Compared to the direct digestion of crude plasma, a lower amount of pepsin and a shorter digestion time were required to completely digest the purified IgG to F(ab′)₂. Complete digestion of purified IgG to F(ab′)₂ was achieved at a pepsin/IgG (w/w) ratio of 5:45 with preservation of structure and potency. Finally, purification of F(ab′)₂ was accomplished by a combination of protein A affinity chromatography and ultrafiltration with a 50-kDa nominal molecular weight cut-off membrane. The new process resulted in 68.9 ± 0.6 (%) total recovery of F(ab′)₂ and a F(ab′)₂ product of high potency.     

Ureterovaginal fistula: A complication of a vaginal foreign body

Objective

To know the diagnostic tools and proper management of ureterovaginal fistula following neglected vaginal foreign body in order to achieve optimal outcome.

Generation of human interferon gamma and tumor Necrosis factor alpha chimeric TNT-3 fusion proteins

Studies have shown that cytokines can effectively treat solid tumors by a direct cytotoxic effect as well as by immunomodulation. Both human interferon gamma (IFNgamma) and tumor necrosis factor alpha (TNFalpha) have been used to treat a variety of colon carcinoma cell lines and tumors in patients. These cytokines, however, are dose limited by their toxicity and fast clearance rates when given intravenously. To improve their therapeutic value, we now report on the generation of two new fusion proteins consisting of human IFNgamma and TNFalpha genetically linked to the C-terminal portion of chTNT-3, a monoclonal antibody (MAb), which targets human solid tumors by binding to intracellular antigens exposed in degenerating cells associated with tumor necrosis. In vitro characterization studies demonstrate that both the IFNgamma and TNFalpha fusion proteins are able to maintain their binding affinity to antigen as well as their direct cytotoxic effect and immunomodulatory functions. When both fusion proteins are combined at optimal doses, they demonstrate a 30% direct cellular cytotoxicity of human colon carcinoma cells of which approximately 14% can be attributed to apoptosis. In vivo, these agents were studied for their pharmakocinetic clearance rates and their ability to target human colon carcinomas heterotransplanted in nude mice. The results of these studies show that, compared with chTNT-3 parental antibody, both fusion proteins have a substantially shorter whole body half-life, yet are able to target tumor in a similar manner. As each of these fusion proteins are cleared from the circulation and normal tissues, tumor-to-normal-tissues ratios rise demonstrating the retention of these reagents in tumor. The generation of long-acting and targeted human IFNgamma and TNFalpha antibody fusion proteins will enable investigators to study the role of these potent immunostimulatory cytokines in the treatment of human solid tumors.     

A complicated urethrovaginal fistula following an inappropriate urethral diverticulum management

Objective

Urethral diverticulum is uncommon, therefore appropriate evaluation, preoperative planning and counseling must be done in order to make correct diagnosis and prevent complications.