土壤微生物多样性研究方法的进展

对土壤微生物的4类研究方法即:传统微生物培养法、微生物标记物法、BIOLOGGN微平板法和微生物分子生物技术方法及其应用特点进行了简要的评述和分析,旨在通过比较寻求能够揭示土壤微生物群落结构的最佳方法。分子生物技术方法与传统研究方法等相结合将是大力推动土壤微生物研究的有效方式。     

Development of biomarkers inAporrectodea caliginosa andLumbricus rubellus for detecting exposure to benzo[a]pyrene

Objectives and Methods. The sensitivity of two biomarkers, the neutral red retention assay (NRRA) and cytochrome P450 were evaluated in the earthwormAporrectodea caliginosa for their potential to detect exposure to benzo[a]pyrene (BaP). Cytochrome P450 was also evaluated in the earthwormLumbricus rubellus, and measured using the substrate ethoxycoumarin. Optimal assay conditions (pH, and temperature) were determined, followed by exposure of earthworms to 20 mg/kg BaP (a typical concentration at contaminated sites in New Zealand). Ethoxycoumarin-O-dealkylase (ECOD) activity was measured after 7, 14, and 21 days exposure. The NRRA was evaluated in earthworms exposed to 0.2, 20, and 100 mg BaP/kg, and biomarker responses were compared with effects on body weight. Results and Discussion Benzo[a]pyrene failed to induce ECOD activity in either earthworm species, and therefore it is not useful as a biomarker of BaP exposure and was not evaluated further. In all cases, the NRRA was significantly affected in the absence of any effects on earthworm body weight, indicating that this assay can detect exposure to BaP at a range of concentrations comparable to those found at contaminated sites. The NRRA should be linked to reproductive endpoints, then it can be used as an early warning indicator of adverse effects. Establishing biomarker stability under environmental conditions is an important step in biomarker development. Therefore, the effects of soil type (sandy soil, silt loam, and a clay soil), moisture content (15–30%), and temperature (5–20°) on the NRRA were determined. In all cases, there was no effect on the NRRA, indicating that this assay is very stable under varying environmental conditions. Conclusion and Outlook In conclusion, cytochrome P450 activity does not appear to be a useful indicator of PAH exposure in eitherA. caliginosa or L.rubellus, and due to the inherently low activity, it is not suitable as a routine biomarker for detecting environmental contamination by these compounds. In contrast, the NRRA in the earthworm A.caliginosa is a promising indicator of PAH exposure at the concentrations likely to be found in contaminated sites in New Zealand, and therefore has potential for evaluating these contaminated sites. If the NRRA can be linked to ecologically important life-cycle endpoints, such as reproduction, then it could be used as an early warning indicator of adverse effects at contaminated sites, i.e., by measuring biomarker responses in earthworms from a ‘contaminated area’ and comparing these with earthworms from a matched control area.     

利用噬菌体随机肽库筛选胰腺癌特异性血清标志物

目的探讨利用噬菌体随机肽库筛选胰腺癌特异性血清标志物的可行性。方法纯化40例正常人和40例首次确诊的胰腺癌患者的血清免疫球蛋白,进行3轮噬菌体随机肽库的亲和筛选,并随机挑选20个克隆进行阳性鉴定,对阳性克隆进行DNA测序和对比分析。结果通过正常对照的阴性排除和3轮亲和筛选,噬菌体随机12肽回收率提高了500倍。随机挑选的20个克隆中有10个阳性克隆,经DNA测序分析获得4条多肽。阳性克隆的敏感性为100%,特异性为95%,阳性预测值为95.2%,阴性预测值为100%,准确性为97.5%。结论利用噬菌体随机肽库能够成功筛选出胰腺癌患者特异性血清标志物。     

Thymidine kinase assay in canine lymphoma

The aim of the study was to evaluate if thymidine kinase (TK) correlated with duration of first remission (DFR) or survival in dogs with lymphoma and if initial TK levels correlated with stage and substage; and also to assess if TK level at diagnosis correlated with immunophenotype. TK was assayed in 73 dogs with treatment naïve lymphoma, then again after treatment; 47% had an initial TK above the reference interval. Dogs with B‐cell lymphoma had higher initial TK activities than dogs with T‐cell lymphoma. TK levels were not higher in dogs with higher stage disease and TK activity prior to treatment was not associated with DFR or survival. Where TK was elevated at diagnosis, it fell into the reference range during remission. TK was normal in 53% dogs at diagnosis, which is higher than previously reported. Further studies are warranted to assess the utility of TK in dogs with lymphoma.     

Plasma haptoglobin and immunoglobulins as diagnostic indicators of deoxynivalenol intoxication

This study aimed to discover potential biomarkers for dioxynivalenol (DON) intoxication. B6C3F1 male mice were orally exposed to 0.83, 2.5 and 7.5 mg/kg body weight (bw) DON for 8 days and the differential protein expressions in their blood plasma were determined by SELDI - Time-of-Flight/Mass Spectrometry (TOF/MS) and the immunoglobulins (Igs) G, A, M and E in the serum were investigated. 11.7 kDa protein was significantly highly expressed according to DON administration and this protein was purified by employing a methyl ceramic HyperD F column with using optimization buffer for adsorption and desorption. The purified protein was identified as a haptoglobin precursor by peptide mapping with using LC/Q-TOF/MS and MALDI-TOF/MS and this was confirmed by western blotting and ELISA. IgG and IgM in serum were decreased in a dose-dependent manner and IgA was decreased at 7.5 mg/kg bw DON administration, but the IgE level was not changed. To compare the expressions of haptoglobin and the Igs patterns between aflatoxin B1 (AFB1), zearalenone (ZEA) and DON intoxications, rats were orally administered with AFB1 1.0, ZEA 240 and DON 7.5 mg/kg bw for 8 days. Haptoglobin was increased only at DON 7.5 mg/kg bw, while it was slightly decreased at ZEA 240 mg/kg bw and it was not detected at all at AFB1 1.0 mg/kg bw. IgG and IgA were decreased by DON, but IgG, IgA, IgM and IgE were all increased by AFB1. No changes were observed by ZEA administration. These results show that plasma haptoglobin could be a diagnostic biomarker for DON intoxication when this is combined with examining the serum Igs.     

Serum biomarker profiling in cancer studies: a question of standardisation?

Companion animals are exposed to similar environmental conditions and carcinogens as humans. In some animal cancers, there also appears to be the same genetic changes associated as in humans. However, little work has been carried out in cancer biomarker identification in animals. The recent dramatic advances in molecular medicine, genomics, proteomics and translational research will allow biomarker identification, which may provide the best strategies for veterinarians and clinicians to combat disease by early diagnosis and administration of effective treatments. Proteomics may have important applications in cancer diagnosis, prognosis and predictive clinical outcome that could directly change clinical practice by affecting critical elemen‐ts of care and management. This review summarizes the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality. In this review, we will discuss the available proteomic technologies and their limitations, and highlight the key areas of research and how they have been used to discover cancer biomarkers. The principles described here are equally applicable to human and animal disease, but implementation of ‘omic’ technologies requires stringent guidelines for collection of clinical material, the application of analytical techniques and interpretation of the data.