Peroxynitrite modified DNA may be an antigenic trigger for antibodies in various cancers of gynecologic origin

This study aimed towards probing the role of peroxynitrite damaged human DNA (ONOO^–-DNA) in the induction of circulating antibodies in certain cancers of gynecologic origin. We have compared the binding specificity of DNA isolated from the lymphocytes of cancer patients with that of the experimentally modified DNA. Also, the induced anti-ONOO⁻-DNA antibodies have been used to probe oxidative damage in the DNA isolated from cancer patients. Human placental DNA was modified with peroxynitrite (ONOO⁻) and analyzed by ultraviolet (UV) and fluorescence spectroscopy, gel electrophoresis, thermal denaturation profile, etc. Antibodies against modified DNA were induced in experimental animals. Specific binding of the antibodies was evaluated by ELISA and band shift assay. 91 cancer patients were selected and grouped according to the type of cancer. Specific binding characteristics of circulating autoantibodies (IgG) were determined by competitive-inhibition ELISA, using different inhibitors. Maximum inhibition of antibody activity by ONOO⁻-DNA reflected specific recognition of modified epitopes by cancer IgG. This shows generation of neo-epitopes on DNA, upon modification with ONOO⁻, that are recognized by cancer IgG. Our results indicate epitope sharing between the DNA isolated from cancer patients and the in-vitro modified ONOO⁻-DNA. The possible role of nitrosative stress in the gynecologic oncology has been discussed.     

A superoxide anion generator, pyrogallol, inhibits the growth of HeLa cells via cell cycle arrest and apoptosis

We investigated the in vitro effects of pyrogallol on cell growth, cell cycle regulation, and apoptosis in HeLa cells. Pyrogallol inhibited the growth of HeLa cells with an IC(50) of approximately 45 microM. Pyrogallol induced arrest during all phases of the cell cycle and also very efficiently resulted in apoptosis in HeLa cells, as evidenced by flow cytometric detection of sub-G1 DNA content, annexin V binding assay, and DAPI staining. This apoptotic process was accompanied by the loss of mitochondrial transmembrane potential (DeltaPsi(m)), Bcl-2 decrease, caspase-3 activation, and PARP cleavage. Pan-caspase inhibitor (Z-VAD) could rescue some HeLa cells from pyrogallol-induced cell death, while caspase-8 and -9 inhibitors unexpectedly enhanced the apoptosis. When we examined the changes of the ROS, H(2)O(2) or O(2)(*-) in pyrogallol-treated cells, H(2)O(2) was slightly increased and O(2)(*-) significantly was increased. In addition, we detected a decreased GSH content in pyrogallol-treated cells. Only pan-caspase inhibitor showing recovery of GSH depletion and reduced intracellular O(2)(*-) level decreased PI staining in pyrogallol-treated HeLa cells, which indicates dead cells. In summary, we have demonstrated that pyrogallol as a generator of ROS, especially O(2) (*-), potently inhibited the growth of HeLa cells through arrests during all phases of the cell cycle and apoptosis.     

Bioactivity-Guided Isolation of Anticancer Agents from Bauhinia Kockiana Korth

Background

Flowers of Bauhinia kockiana were investigated for their anticancer properties.

Methods

Gallic acid (1), and methyl gallate (2), were isolated via bioassay-directed isolation, and they exhibited anticancer properties towards several cancer cell lines, examined using MTT cell viability assay. Pyrogallol (3) was examined against the same cancer cell lines to deduce the bioactive functional group of the phenolic compounds.

Results

The results showed that the phenolic compounds could exhibit moderate to weak cytotoxicity towards certain cell lines (GI₅₀ 30 – 86 µM), but were inactive towards DU145 prostate cancer cell (GI₅₀ > 100 µM).

Conclusion

It was observed that pyrogallol moiety was one of the essential functional structures of the phenolic compounds in exhibiting anticancer activity. Also, the carboxyl group of compound 1 was also important in anticancer activity. Examination of the PC-3 cells treated with compound 1 using fluorescence microscopy showed that PC-3 cells were killed by apoptosis.     

In Vitro Anti-Oxidative Activities of the Various Parts of Parkia Biglobosa and GC-MS Analysis of Extracts with High Activity

The anti-oxidative activities of sequentially extracted solvent fractions of different parts of P. biglobosa were evaluated in a series of in vitro assays. Our findings indicated that all extracts had electron donating and free radical scavenging activities. But the ethanol (EtOH) extracts from all the parts demonstrated more promising anti-oxidative effects in these experimental models. Apart from the aqueous extracts of the stem bark and leaves, all other extracts exhibited hydroxyl radical scavenging (HRS) activity but the ethyl acetate (EtOAc) extract of the stem bark and EtOH extracts of the root and leaves possessed more powerful HRS activity than other corresponding extracts in the parts. Further, nitric oxide (NO) inhibition activities were observed in all the extracts except the EtOAc extract of the stem bark which showed pro-oxidative activity. However, the EtOH extract of the stem bark and root as well as the EtOAc extract of the leaves displayed more potent anti-NO activity than other extracts in the parts. The GC-MS analysis of the EtOH extracts revealed that the most abundant phytochemicals are pyrogallol derivatives. Data from this study suggest that the EtOH extracts from different parts of P. biglobosa contained potent anti-oxidative agents and pyrogallol could be the main bioactive constituent.     

碳糊电极吸附阳极溶出伏安法测定血清中铜

利用液体石蜡作粘合剂制成碳糊电极(CPE),根据铜与邻苯三酚红(PGR)生成的络合物在CPE上的吸附特性,建立了测定铜的吸附溶出伏安法。考察了最佳底液条件,共存离子的影响,测定了电活性络合物的组成,探讨了电极反应机理。二次导数溶出峰电流与Cu2+浓度在2.0×10-9molL～4.0×10-7molL范围内呈线性关系。检出限为1×10-9molL。方法应用于人血清样中铜的测定,取得了满意的结果。     

健康大学生红细胞超氧化物歧化酶活性

用改良的邻苯三酚自氧化法检测了117例健康大学生红细胞超氧化物歧化酶(SOD)活性。1个酶单位是指在25℃、pH8.2和325nm紫外光分光光度法测定时,每ml反应液中每分钟抑制邻苯三酚自氧化速率达50％的酶量。健康大学生红细胞SOD活性为533.48±83.81单位/ml红细胞,无显著的性别差异。本法批内及批间变异分别为3.14％和5.27％。作者认为以每ml红细胞中SOD活力单位表示较为简便、正确。     

Hepatoprotective effect of New Livfit, a polyherbal formulation, is mediated through its free radical scavenging activity

The effect of New Livfit, a polyherbal formulation, was studied on pyrogallol-induced hepatotoxicity in rats. Administration of pyrogallol 100 mg/kg, i.p. caused a significant increase in liver enzymes as well as a significant increase in lipid peroxidation. Simultaneous administration of oral New Livfit and pyrogallol prevented these changes in hepatic damage. The results of the study showed that New Liv fi t(R) exerted a hepatoprotective effect against pyrogallol induced liver toxicity, which was mediated through its free radical scavenging property.     

Development of a new method for measuring total urinary protein using acid violet 6B pigment

We have developed a new method for measuring total urinary protein using acid violet 6B (AV6B) pigment. Nine purified components of human urinary proteins and urine samples collected randomly from 123 diabetic outpatients were used. There were 62, 36, and 25 cases of prenephropathy, early nephropathy, and overt nephropathy, respectively. All samples were measured by Coomassie brilliant blue G 250 (CBB), pyrogallol red-molybdate (PRM), and AV6B methods using an optical photometer. In healthy subjects, the major components of urinary proteins, such as gamma-globulins, IgG, IgA Tamm-Horsfall protein, and transferrin, the reactivity values of the AV6B and PRM methods were similar. The CBB method was the least sensitive of the three methods. In the urine samples from diabetic patients, the urinary protein values measured by the AV6B method were higher than those measured by the CBB method in the prenephropathy stage. The values obtained by the AV6B method (y) correlated well with those from the CBB method (x) (y=1.243x+3.61, r=0.904). When the values from the AV6B method (y) were compared to those from the PRM method (x), correlation was low (y=1.406x-29.15, r=0.786). In conclusion, the AV6B method was more useful than the CBB and PRM methods for low levels of urinary protein.     

红岩草中三种成分的体外抑菌实验及组方优选研究

目的：通过红岩草中连苯三酚、氢醌和没食子酸甲酯等3种成分的体外抑菌作用,验证红岩草的抑菌活性,同时为其成分组方提供依据。方法：采用常量稀释法,测定连苯三酚、氢醌、没食子酸甲酯对导致奶牛乳房炎5种主要致病菌的体外抑菌效果;采用正交试验方法优选3种成分的最佳配比并对其进行验证。结果：连苯三酚对金黄色葡萄球菌、表皮葡萄球菌、大肠杆菌、无乳链球菌和停乳链球菌的最低抑菌浓度分别为160 mg.L-1、80 mg.L-1、1 280 mg.L-1、640 mg.L-1和640 mg.L-1,氢醌对5种细菌的最低抑菌浓度分别为640 mg.L-1、80 mg.L-1、〉1 280 mg.L-1、80 mg.L-1和80 mg.L-1,没食子酸甲酯对5种细菌的最低抑菌浓度分别为12 800 mg.L-1、〉1 280 mg.L-1、640 mg.L-1、〉1 280 mg.L-1和〉1 280 mg.L-1。正交试验结果直观分析显示连苯三酚、氢醌、没食子酸甲酯对于金黄色葡萄球菌、表皮葡萄球菌、大肠杆菌和无乳链球菌的抑菌效果为连苯三酚〉氢醌〉没食子酸甲酯;对于停乳链球菌的抑菌效果为氢醌〉没食子酸甲酯〉连苯三酚。方差分析显示,3种成分对金黄色葡萄球菌、表皮葡萄球菌和大肠杆菌,因素水平间存在显著性差异（P〈0.05）,而对于无乳链球菌和停乳链球菌,因素水平间无显著性差异。结论：氢醌对奶牛乳房炎主要致病菌有较好的体外抑菌作用,3种成分的最佳组方为A3B3C2。     

焦性没食子酸对3种不同水生生物的毒性作用

目的研究焦性没食子酸对水生生物的毒性作用。方法以蛋白核小球藻(Chlorella pyrenoidosaChick)、斜生栅藻(Scenedesmus obliquus)和多刺裸腹溞(Moina macrocopa)作为受试对象,探讨不同浓度的焦性没食子酸(pyrogallol)的急性毒性效应。结果随焦性没食子酸浓度的增加藻细胞密度和溞数量明显下降,呈现剂量-效应关系;焦性没食子酸对蛋白核小球藻和斜生栅藻半抑制效应浓度(EC50)分别为19.7mg/L和19.8mg/L,对多刺裸腹溞24h和48h半抑制效应浓度分别为18.25mg/L和13.96mg/L。结论焦性没食子酸对蛋白核小球藻、斜生栅藻和多刺裸腹溞均属于中毒。