Study of neonatal exposure to androgenic endocrine disruptors, testosterone and dihydrotestosterone by normal-phase HPLC

Neonatal exposure to androgen induces developmental abnormalities in the male reproductive system. To investigate whether neonatal exposure affects spermatogenesis in juvenile and pubertal testis, Sprague-Dawley rat pups were given androgen or various androgenic endocrine disruptors by a single injection on the day of birth at concentrations ranging between 4 mm to 200 mm, and sacrificed on day 21 (juvenile) or 50 (puberty). The testes were weighed and examined histologically at each stage. Further, the metabolites of steroidogenesis were analyzed using normal-phase high-performance liquid chromatography. Neonatal exposure significantly reduced testis weights and steroidogenesis of juveniles. Neonatal exposure to testosterone and dihydrotestosterone still suppressed pubertal steroidogenesis, although testis weight was completely restored during puberty.     

Sodium dodecyl sulfate versus acid-labile surfactant gel electrophoresis: comparative proteomic studies on rat retina and mouse brain

A long-chain derivative of 1,3-dioxolane sodium propyloxy sulfate, with similar denaturing and electrophoretic properties as SDS, and facilitated protein identification following polyacrylamide gel electrophoresis (PAGE) for Coomassie-stained protein bands, has been tested. Comparative acid-labile surfactant/sodium dodecyl sulfate two-dimensional (ALS/SDS 2-D)-PAGE experiments of lower abundant proteins from the proteomes of regenerating rat retina and mouse brain show that peptide recovery for mass spectrometry (MS) mapping is significantly enhanced using ALS leading to more successful database searches. ALS may influence some procedures in proteomic analysis such as the determination of protein content and methods need to be adjusted to that effect. The promising results of the use of ALS in bioanalytics call for detailed physicochemical investigations of surfactant properties.     

Liquid chromatography‐tandem mass spectrometric method for determination of E121 in mouse plasma and its application to pharmacokinetics

A rapid LC‐MS/MS method for quantification of an enaminone analog, E121 in mouse plasma using E118 as an internal standard (IS) has been developed and validated. The analyte was analyzed on C₁₈ column using a mobile phase of acetonitrile/methanol/ammonium acetate/formic acid (60:20:20:0.025, v/v/v/v) at a flow rate of 0.25 mL/min. Quantitation was achieved using ESI+ interface, employing MRM mode at m/z 308>262 and 222>194 for E121 and IS, respectively. The calibration standards were linear over a range of 0.10–20 μg/mL (r²>0.99) with an LLOQ of 0.1 μg/mL (RSD%; 11.4% and bias%; 9.5%). Intra‐ and inter‐run precision of E121 assay ranged from 3.7 to 10.9% with accuracy (bias) that varied between ?10.0 and 12.0%, demonstrating good precision and accuracy. Recoveries of E121 and the IS from plasma were above 80%. Stability of E121 in plasma showed that the analyte was stable under various conditions. The matrix effect study showed a lack of effect. The applicability of the developed method was demonstrated by measuring E121 in mouse plasma samples following intraperitoneal administration of various doses ranging from 10 to 100 mg/kg and this study demonstrates that E121 exhibits linear kinetics in the dose range studied.     

Dietary genistein enhances phosphorylation of regulatory myosin light chain in the myocardium of ovariectomized mice

There is evidence that isoflavones, such as genistein, can directly or indirectly improve lipid profile and lower blood pressure and hence exert cardiovascular protection. It is further believed, that genistein attenuates vascular contraction and thus vascular tone and blood pressure through altering the phosphorylation of the regulatory myosin light chain (MLC) probably via the myosin light chain kinase (MLCK) or the RhoA pathway. However, the direct role of genistein in the myocardium is poorly reviewed. In this study, we investigated the impact of genistein on the cardiac proteome in ovariectomized female mice using a 2DE-MS approach. Dietary genistein intake considerably changed the abundance of several cytoskeletal and contractile proteins and enhanced the phosphorylation of MLC. The MLC phosphorylation was mediated through increased abundance of MLCK and inhibition of myosin light chain phosphatase latest known to be inversely regulated by RhoA. Contrary to others, in our model genistein did neither inhibit the cardiac MLCK, nor the cardiac RhoA pathway in vivo.     

液相等电聚焦技术结合液相色谱-质谱联用技术分析鼠肝全蛋白

利用液相等电聚焦预分离技术结合液相色谱-质谱(LTQ-Orbitrap)联用技术,研究了C57小鼠肝脏的蛋白质表达谱.质谱分析结果采用Max Quant1.4.1.2软件搜索数据库,共鉴定出3474个蛋白(2个以上唯一肽段).用DAVID在线工具、GO分类工具和IPA软件对鉴定蛋白进行生物信息学分析,单独发现832个新蛋白.研究结果表明,通过基于等电点和亲疏水性的两维分离,提高了质谱鉴定蛋白数,可以鉴定出更多低丰度蛋白.     

兴国红鲤精巢发育的研究

以289尾兴国红鲤(Cyprinus carpio var. singuonensis)雄体为材料,研究了精巢在第一次性周期内的发育。雄鱼3月龄,精巢发育为第Ⅰ期,4月龄发育为第Ⅱ期,4～6月龄发育为第Ⅲ期,8～10月龄发育为第Ⅳ期,11～12月龄发育为第Ⅴ期,性成熟期为1周龄。性成熟后精巢发育的周年变化表明;     

HPLC quantification of the HIV-1 protease inhibitor saquinavir in brain and testis of mice

A rapid, reliable HPLC method with UV detection (240 nm) was developed and validated for quantitation of saquinavir in mice brain and testis. Saquinavir and the internal standard were isolated from homogenized tissue matrices using liquid-liquid extraction procedure and were then analyzed using an isocratic mobile phase by reversed-phase liquid chromatography. The lower limit of quantification was 50 ng/g for both brain and testis. A linear dynamic range of 50-5000 ng/g for both brain and testis was established. This HPLC method was validated with between-batch precision of 0.5-4.4 and 1.5-5.5% for brain and testis, respectively. The between-batch accuracy was 94.7-105.9% and 97.5-105.0% for brain and testis, respectively. The present method was applied for tissue distribution studies of the novel drug delivery systems of saquinavir in mice.     

Discovery of lung squamous carcinoma biomarkers by profiling the plasma peptide with LC/MS/MS

Biomarkers can be used for the screening and clinical diagnosis of cancer, and peptidomics approach has been proven successful in the research of biomarkers. To develop better peptidomic technologies for fast, accurate, and reliable detection of peptides biomarkers for lung cancer, we have improved the procedures of blood collection to minimize the degradation of the blood proteins and optimize the extraction of peptidome peptides from plasma samples based on acetonitrile precipitation associated with size exclusion chromatography (SEC). Studies show that squamous cell carcinomas are found to express CAGE1, SPAT9 and TEX28 genes at significantly higher rates, and the results suggest that as tumors progress, the level of CAGE1, SPAT9 and TEX28 genes are likely to increase and lead to immunization. This suggests a potentially important therapeutic method for cancer testis-based cancer vaccines.     

Crocetin Mitigates Irradiation Injury in an In Vitro Model of the Pubertal Testis: Focus on Biological Effects and Molecular Mechanisms

Infertility is a potential side effect of radiotherapy and significantly affects the quality of life for adolescent cancer survivors. Very few studies have addressed in pubertal models the mechanistic events that could be targeted to provide protection from gonadotoxicity and data on potential radioprotective treatments in this peculiar period of life are elusive. In this study, we utilized an in vitro model of the mouse pubertal testis to investigate the efficacy of crocetin to counteract ionizing radiation (IR)-induced injury and potential underlying mechanisms. Present experiments provide evidence that exposure of testis fragments from pubertal mice to 2 Gy X-rays induced extensive structural and cellular damage associated with overexpression of PARP1, PCNA, SOD2 and HuR and decreased levels of SIRT1 and catalase. A twenty-four hr exposure to 50 μM crocetin pre- and post-IR significantly reduced testis injury and modulated the response to DNA damage and oxidative stress. Nevertheless, crocetin treatment did not counteract the radiation-induced changes in the expression of SIRT1, p62 and LC3II. These results increase the knowledge of mechanisms underlying radiation damage in pubertal testis and establish the use of crocetin as a fertoprotective agent against IR deleterious effects in pubertal period.