Comparative identity and homogeneity testing of the mtDNA HV1 region using denaturing gradient gel electrophoresis

A denaturing gradient gel electrophoresis (DGGE) assay has been developed for comparative identity and homogeneity testing of the mtDNA HV1 region. A total of 49 pairs of sequences, each pair differing by a single unique polymorphism, were tested to verify the reliability of the assay. Discrimination between all pairings was achieved as judged by the resolution of the mismatch-containing heteroduplexes from the fully base-paired homoduplexes. In all but two pairings, resolution of the fully base-paired homoduplexes was also obtained. Sequence pairs differing by multiple polymorphisms were also tested and resulted in a greater separation between the homo- and heteroduplexes. Additional information derived from the technique includes the identification of co-amplifying contaminating or heteroplasmic samples in the independent samples lanes. Thirteen heteroplasmic samples, six at positions distinct from those analyzed in the pairwise comparison study, were analyzed and the heteroplasmic positions identified unambiguously by sequencing the excised bands. The technique constitutes a conceptually simple, accurate, and inexpensive test for determining whether two sequences match within the mtDNA HV1 region, while providing a more definitive control for the identification of co-amplifying contaminating or heteroplasmic sequences than is presently available.     

Haptoglobin phenotyping of bloodstains by horizontal electrophoresis on a compact polyacrylamide gradient gel

A method is described for phenotyping haptoglobin by horizontal electrophoresis on a small polyacrylamide gradient gel. This method employs the same apparatus used in the separation of many red cell enzyme phenotypes and thereby eliminates the necessity for specialized vertical electrophoresis equipment.     

The application of immunoblotting to the phenotyping of haptoglobin.

An immunoblotting method for phenotyping haptoglobin in serum and bloodstains has been developed. Haptoglobin isoproteins were separated by polyacrylamide gradient gel electrophoresis and then transferred to nitrocellulose by electroblotting. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm thick gels. Nitrocellulose blots were developed by double antibody enzyme immunoassay. The detection limit for serum and bloodstains was improved 16 times compared to conventional staining using O-tolidine. The method could detect haptoglobin phenotypes from 0.001 microliter of whole blood. This detection limit is approximately 8 times lower than that of group specific-component analysis by immunoblotting.     

单细胞凝胶电泳检测大鼠死后肝细胞核DNA降解

目的应用单细胞凝胶电泳技术(SCGE)检测大鼠死后肝细胞核DNA降解规律,分析与死亡时间的关系,为早期死亡时间的推断提供新的方法。方法在大鼠死后30h内,每隔3h取肝组织样本进行单细胞凝胶电泳,用共聚焦显微镜摄取彗星图像,应用彗星图像分析软件(IM I1.0)进行图像分析,并作统计学分析。结果死后大鼠的肝细胞在电泳图像上出现明显的彗星形拖尾,其尾长(TL)、尾矩(TM)在一定的时间范围内(0~18h)随死亡时间的延长而逐渐增大,二者均与死亡时间(PM I)呈现一定的相关回归关系。结论单细胞凝胶电泳技术可应用于早期死亡时间的推断。     

签字笔书写笔痕特征的实验分析

目的为准确掌握签字笔书写笔痕特征的变化规律，以求将其运用于案件鉴定。方法通过对同一人执不同签字笔、不同人执同一支签字笔及同一人执同一支签字笔书写笔痕分别进行实验分析。结果证实前二种情况下笔痕特征具有相互区别的特定性，后一种情况下笔痕特征具有重复再现的稳定性。     

Investigation of the short-time autolysis of rat hearts by means of SDS. Polyacrylamide gel electrophoresis and electron microscopy.

The short-time autolysis of hearts was regarded as a model of ischaemic heart failure. Therefore, isolated rat hearts were subjected to 30--120 min autolysis in a Locke solution at 37 degrees C. Electron microscopic examinations and myofibrillar preparations were made from the autolysed heart ventricles. The myofibrillar proteins were resolved by SDS-polyacrylamide gel electrophoresis. After 30 min autolysis the amount of a protein of 192,000 daltons greatly increased. At the same time on the electron micrographs the focal destruction of filament destruction on the A filament area and the mitochondrial structure altered too. After 60 min autolysis another protein of 36,400 daltons appeared. On the electron micrographs the focal desintegration of Z membranes and the focal destruction of I filaments can be observed. After 120 min autolysis further proteolytic products could not be detected by gel electrophoresis but on the electron micrographs the destruction of Z membranes and I filaments became more pronounced.     

美国商业方法 专利制度正当性分析

1998年,美国联邦巡回上诉法院(CAFC)就“StateStreet Bank Trust Co.v.Singature Financial GroupCo.”(以下简称“State Street Bank”案)一案做出判决,肯认了商业方法的可专利性,由此引发了学界对于商业方法专利的关注。 (一)“State Street Bank”案 与商业方法是可专利主题 该案被告拥有美国专利第5793106号专利,即“轴幅式金融服务配置资料处理系统”(Data Processing forHub and Spoke Financial Services Configuration)专利。该专利的用途在于管理与计算由数个共同基金所组成的合伙关系。由于该发明所涉及的计算十分复杂,因此必须通过电脑等设备完成。在专利授权书中,该电脑被描述     

美国知识产权保险制度管窥

美国的知识产权保险分为知识产权执行保险(IPEnforcementInsurance)和知识产权侵权保险(IPInfringementInsurance)。该保险所保护的主体范围非常广泛,既包括潜在的侵权人,也包括潜在的受害人。虽然知识产权保险制度具有许多优点,但缔结保险合同的保费过高,同时由于大企业的拖延战术,也大大地削弱了该制度的功效。因此,我国企业在知识产权诉讼中,应当充分重视和解与诉讼中间裁决的作用,采取正确的措施渡过难关。     

Determination of the age of bloodstains using immunoelectrophoresis.

The technique of immunoelectrophoresis was used to determine the age of bloodstains. The immunoelectrophoretic patterns (IEP) of bloodstains ranging from 15 days to one year old were obtained by the use of high titer anti-whole human serum. The IEPs revealed gradual disappearance of beta-globulins and gamma-globulin with increase in the age of bloodstains. A comparative study of the IEP of normal human serum with those of the experimental bloodstains showed the absence of some of the corresponding proteins. The absence of a particular serum protein in the IEP of a given bloodstain will indicate the age of that bloodstain.     

Genetic profile of the Madeira Archipelago population using the new PowerPlex16 System kit

Allele and haplotype frequencies of 17 chromosome STR loci, 15 of them included in the kit PowerPlex 16 System from Promega, were determined in a sample of unrelated males from Madeira and Porto Santo Islands. PowerPlex 16 includes STRs not studied before in the Madeira population. The-kit includes two new allele markers (Penta D and Penta E), which proved to be extremely useful for paternity testing (PD = 0.952 and 0.977, respectively). The study revealed that the Madeira population does not differ from that of the north Portugal. Nevertheless, some loci presented alleles found in sub-Saharan and North European populations which were not reported so far in Portugal.     

论举证时限的裁量空间

举证时限制度设置的目的应为确立与法定顺序主义相适应的＂双阶型＂审判流程。证据适时提出主义赋予了法官过大的裁量权,与设置我国举证时限制度的目的相去甚远。我国民事审判流程的革新本身是想朝着＂双阶型＂审判流程而去,却深受民法法系国家证据随时提出主义的影响,淡化了驱动＂双阶型＂审判流程的法定顺序主义,最终陷入＂裁量陷阱＂。《举证时限通知》的进步在于对法官在举证时限制度运行中过度裁量权的限缩。举证时限制度的合目的运行不仅需要程序技术的进步,更需要程序伦理的支撑与推动。《举证时限通知》的进步仅仅是程序技术方面的进步。     

快速溶剂萃取-凝胶色谱净化-LC／MS／MS相结合测定血中的杀参毒素类农药

目的建立血中杀参毒素类农药残留的快速分析方法。方法取一定量的Al2O3放入萃取池中做吸附剂,通过快速溶剂萃取仪萃取血液中杀参毒素类农药,同时达到在线净化的效果,后运用凝胶色谱净化联用仪进一步净化浓缩到2ml,提取液经LC／MS／MS进行检验。结果杀虫双在1．2×10g／ml～1．2×10^-6g／ml范围内线性关系良好,检出限为0．04ppb,杀虫眯为1．2×10^-6g／ml～7．2×10g／ml,检出限为0．34ppm。2种杀参毒素农药的平均回收率为89％、90％,相对标准偏差为1．9％、3％。结论整个方法简便、快速、准确、重现性好、灵敏度高,杂质干扰少。     

美国违宪审查的历史探渊

美国违宪审查的思想渊源至少可以追溯到英国的爱德华·柯克爵士,柯克的著作将违宪审查的种子撒播于北美之后,新大陆独特的环境哺育了它。特别是在独立战争前夕,它成为北美人民对抗英国政府统治的有效武器。美国独立后,联邦宪法的颁布、批准宪法的论战以及马伯里案之前的违宪审查实践为美国违宪审查制度的建立奠定了思想、制度和先例方面的坚实基础。尽管美国违宪审查制度的最终确立离不开马歇尔大法官在马伯里案中的创造性发挥,但马歇尔只是催生了它,历史女神克利奥对违宪审查的孕育才是违宪审查制度得以产生的关键。     

Development of latent fingerprints on metallic surfaces using electropolymerization processes.

We propose a new process for developing latent fingerprints on metal items, applicable to unfired weapons made of Ergal in particular. The method is based on the presence of fatty acids that are contained in fingerprints and act as an insulator on the surface where fingerprints are to be developed. The process of polymerization occurs on the metal portions left untouched by finger contact. Hence, the developing process results as a negative pattern of the original fingerprint. The reaction consists in the electropolymerization of pyrrole and substituted porphyrins, i.e., tetra (o-aminophenyl) porphyrine: radical-cations are generated on superficial nucleation sites by oxidation of monomer, close to the electrode surface; subsequently, the radical species react with the neutral monomer, which begins to diffuse to the electrode. We have also studied the polymer's morphology by means of SEM and AFM, in order to find a correlation between the reagent to be used and the quality of the enhancement process. These are only preliminary results; however, they show that the suggested method is a new way to increase the rate of success in developing latent fingerprints on metal surfaces. In this regard, it may be considered complementary to other conventional procedures, due to the low costs of the instruments and reagents, and the rapidity and simplicity of the treatment.     

Determination of mouth alcohol using the Dräger Evidential Portable Alcohol System

Drivers suspected of alcohol intoxication are observed for a period of 15 min prior to quantitative breath alcohol testing. This is to preclude the interference of alcohol-based substances such as cough medicine, mouthwash, and breath spray just prior to actual evidential testing. To determine whether a 15 min observation period was necessary when performing evidential breath tests in the field, a mouth alcohol experiment was performed using the Dräger Evidential Portable Alcohol System (EPAS). Five types of alcohol beverages and the effects of expectorating versus swallowing were tested on twenty-five volunteer subjects. Serial measurements of breath and blood alcohol levels were performed at fixed time intervals. All alcohol beverage types gave two sequential measurements within 0.02 g/210 L of each other before 15 min had passed. Fifteen minutes was necessary to ensure there was no residual mouth alcohol. If the 15 min waiting period was not observed, the safety feature of the EPAS requiring two sequential measurements 2 min apart within 0.02 g/210 L would not ensure against mouth alcohol interference.     

The recovery of semen from bathwater using the Evidence Recovery System (ERS)

Sexual offences are under-reported and ascertaining accurate offence numbers is difficult. Any methods which could increase the ability to obtain biological evidence or reduce the additional distress associated with reporting a sexual offence may result in an increase in reporting this crime type. The Evidence Recovery System (ERS) is designed to collect trace evidence, including hairs, fibres and biological evidence, from bath or shower water in a non-invasive manner. Initially, samples of semen were placed in baths filled with water, and washing was simulated using a range of body wash products. The water was then drained through the ERS before its filters were subjected to acid phosphatase testing and haematoxylin and eosin staining of spermatozoa. Recovered spermatozoa were then graded accordingly. Following this, the experiment was repeated with the addition of dirt/dust particulates during the washing stage, to simulate recovery of biological evidence in a more realistic environment. The results showed that spermatozoa considered ‘easy to find’ could regularly be obtained from bathwater using the ERS. It appeared that this recovery was not affected by the presence of different body wash products. When dust/dirt particles were added, the number of spermatozoa recovered increased at two of the evidence collection stages. The difference in recovery was considered to be statistically significant. This study provides evidence to suggest the feasibility of use of the ERS as a method to collect semen evidence from individuals subjected to sexual offences. The recovery of spermatozoa does not appear to be affected by the presence of a body wash, but does appear to be improved when skin cells, hair and other debris are transferred into the water, as would be likely during a bath/shower. Further to this, the possibility of obtaining spermatozoa from the home bath or shower of a victim following a post-offence bathing experience is implied.