猪ERK6基因编码区的克隆及组织表达谱分析

利用RT-PCR方法扩增了猪ERK6基因编码区的序列,将扩增产物与pMD18-T载体连接,构建了重组质粒;重组质粒经PCR、酶切鉴定后进行测序;采用Northern杂交和半定量RT-PCR方法在猪不同部位骨骼肌中分析了ERK6基因转录本的个数、大小及组织表达谱。结果显示,所克隆的猪ERK6基因片段长1 113 bp,含有1个1 104 bp的开放阅读框(ORF),该ORF编码367个氨基酸;该基因与已报道的人ERK6基因核苷酸序列相似性为90%;猪ERK6基因在肌肉组织中只有一个转录本,大小约1.5 kb。此基因在骨骼肌中的表达量最高,心、子宫次之,卵巢最低;而在脂肪、胃、肝、脾、肺、肾、十二指肠和胰腺中未见表达。结果表明,猪ERK6基因与已报道的小鼠和人ERK6基因一样,主要在骨骼中特异表达。     

A Profile of Poverty for Palestinian Refugees in Jordan: the Case of Zarqa and Sukhneh Camps

This is one of few known studies on poverty and poverty relatedissues among Palestinian refugees in Jordan. The camp refugeehouseholds identified in this research represent concrete manifestationsof the ways in which the political and historical legacy translatesinto economic hardship today. Poverty among the refugees inJordan represents a legacy of the refugees’ inabilityto generate sufficient income in order to provide for themselvesthe ‘minimum basic needs’. Therefore, poverty shouldbe understood not only in terms of the social and economic circumstancesof Palestinian camp refugees but also as a particular consequenceof various policies pursued by the state over the past fivedecades, including the Jordanization of public and militarysectors of employment, the provision of public services, andwage determination policy. In this context, the paper examinesthe causes of poverty and considers how various ‘implicit’policies contribute towards luring and ‘locking’a large number of camp refugees in poverty.     

Time burnt away: The impact of heat-induced changes on skeletal age-at-death diagnostic features

This paper investigates the established notion that bone calcination has a major impact on age estimation while low-intensity burns have a mere negligible impact. Few systematic researches have been carried out so far about this topic so the true impact of heat-induced changes on diagnostic age features is mostly unknown.The agreement between pre-burning and post-burning observations of age features was investigated on 51 human skeletons (22 males and 29 females with ages-at-death ranging from 61 to 93 years old) subjected to experimental burns. These skeletons belong to the 21st Century Identified Skeletal Collection housed at the Laboratory of Forensic Anthropology of the Department of Life Sciences, University of Coimbra, Portugal. The Suchey-Brooks method based on the pubic symphysis and the method developed on the auricular surface by Buckberry and Chamberlain (2002) were scrutinized.The Suchey-Brooks method provided better agreement between pre- and post-burn observations than the method from Buckberry and Chamberlain (2002). However, it became clear that heat-induced changes affected both methods regardless of heat intensity since both calcined bones and bones burnt at lower intensities often showed less than perfect agreement. Therefore, this research demonstrates that the analysis of age-at-death can be impaired in burnt bones, even those not subjected to calcination, with clear impact for forensic and archaeological investigations.     

Automated DNA casework workflow: A retrospective study of the first implementation of FIDL at the Netherlands Forensic Institute

FIDL is a fast and automated DNA identification line which represents a series of software solutions automating the process from raw capillary electrophoresis data to reporting. This retrospective study provides insight in the numbers of cases, turnaround time, results compared to the standard workflow and the benefits automation has in a large volume workflow.     

Performance evaluation of the Scent Transfer Unit (STU-100) for organic compound collection and release

The Scent Transfer Unit (STU-100) is a portable vacuum that uses airflow through a sterile gauze pad to capture a volatiles profile over evidentiary items for subsequent canine presentation to assist law enforcement personnel. This device was evaluated to determine its ability to trap and release organic compounds at ambient temperature under controlled laboratory conditions. Gas chromatography-mass spectrometry (GC-MS) analyses using a five-component volatiles mixture in methanol injected directly into a capture pad indicated that compound release could be detected initially and 3 days after the time of collection. Additionally, 15 compounds of a 39-component toxic organic gaseous mixture (10-1000 parts per billion by volume [p.p.b.(v)]) were trapped, released, and detected in the headspace of a volatiles capture pad after being exposed to this mixture using the STU-100 with analysis via GC-MS. Component release efficiencies at ambient temperature varied with the analyte; however, typical values of c. 10% were obtained. Desorption at elevated temperatures of reported human odor/scent chemicals and colognes trapped by the STU-100 pads was measured and indicated that the STU-100 has a significant trapping efficiency at ambient temperature. Multivariate statistical analysis of subsequent mass spectral patterns was also performed.     

LCMS Analysis of Fingerprints,the Amino Acid Profile of 20 Donors

The analysis of amino acids present in fingerprints has been studied several times. In this paper, we report a method for the analysis of amino acids using an fluorenylmethyloxycarbonyl chloride‐derivatization for LC separation and MS detection. We have obtained good results with regard to the calibration curves and the limit of detection and LOQ for the target compounds. The extraction of the amino acids from the substrates used proved to be very efficient. Analysis of the derivatized amino acids enabled us to obtain full amino acid profiles for 20 donors. The intervariability is as expected rather large, with serine as the most abundant constituent, and when examining the total profile of the amino acids per donor, a characteristic pattern can be observed. Some amino acids were not detected in some donors, or fell out of the range of the calibration curve, where others showed a surprisingly high amount of material in the deposition analyses. Further investigations will have to address the intravariability of the amino acid profiles of the fingerprints from donors. By the development of the analytical method and the application to the analysis of fingerprints, we were able to gain insight in the variability of the constituents of fingerprints between the donors.     

SMART:自主研发的混合STR图谱分析系统

目的混合STR图谱的分析是法医遗传学领域的一个难题。目前国内大多数法医DNA实验室依靠人工方法分析混合STR图谱,费时费力,且拆分效果也多有主观性,难以满足日益增长的混合STR图谱分析的需求。本文介绍一套自主研发的混合STR图谱分析系统——SMART(STR Mixture Analysis and Resolution Tools),其能够实现对混合STR图谱的自动化分析,包括拆分混合STR图谱和计算似然比。方法SMART使用的STR峰高的概率模型,考虑了影子峰、降解、基因座特异性扩增效率、峰高变异、插入峰、峰丢失等因素。模型通过比较各个基因型集合拟合当前混合STR图谱的似然值大小来推断最有可能的基因型集合,采用了马尔可夫链蒙特卡洛(MCMC)算法进行求解。结果SMART可应用于不同试剂盒和遗传分析仪产生的数据。经使用ABI-3500XL遗传分析仪和Global Filer试剂盒测试,SMART能够实现对2~5人的混合STR图谱的分析,输出包括混合比例、混合图谱质量、各个贡献者单人的STR基因分型、似然比等多项结果。结论SMART作为一款自主研发的混合STR图谱分析系统,其基本涵盖了国外同类分析系统所具有的功能,技术性能指标达到国际先进水平,能够满足一线法医工作者对于混合STR图谱分析的多样化需求,提高混合STR图谱结果的利用率。     

Using PMCT of Individuals of Known Age to Test the Suchey–Brooks Method of Aging in Victoria,Australia

The estimation of an individual's age at death plays a critical role in the investigation of unidentified human remains. The Suchey–Brooks method, which involves the analysis of degenerative morphological changes in the pubic symphysis, is currently widely used to estimate age in adults. This study tested the applicability of the Suchey–Brooks method on a contemporary adult Australian (Victorian) subpopulation by observing three‐dimensional volume‐rendered postmortem computed tomographic (PMCT) reconstructions of the pubic symphysis of 204 individuals aged 15–100 years. Results showed the method was reliable for females (85% correct allocations) but not for males (67% correct allocations). It is therefore recommended that extreme caution be used when applying the Suchey–Brooks method for estimating age at death of an individual in this subpopulation from PMCT images. The results suggest further investigation into alternate age estimation methods is required.