Application of the hexagon-OBTI test and the RSID blood test for the determination of the post-mortem interval of bone samples

In this serial experiment, five human bones with known post-mortem intervals (PMI) in a soil environment from five different epochs (0.2 to approximately 2000 years) were tested in a blind setup with two established rapid tests for the identification of human blood traces (Hexagon-OBTI test and RSID blood test). Based on previous study results concerning the usability of the Luminol test for the first assessment of the PMI of osseous remains, the question arising was whether those test procedures, which are highly sensitive for the detection of human blood components, could also be used to narrow down the post-mortem interval. Five test series were conducted applying modified standard protocols of the manufactures. The aim was to find out whether with prior reaction steps or a prolonged time of incubation hemoglobin or its metabolites can be dissolved from the bone and positive test results can be achieved dependent on the PMI. Four test series yielded negative results for all bone samples and one test series a uniformly weak positive result. The results indicate that rapid tests based on the detection of blood are not suitable for the determination of the PMI of bone samples despite the modification of the standard protocols. Further thorough research is required to clarify the postmortem degradation of hemoglobin in bones.     

The comparative assessment of the practical value of the currently employed methods for the recognition and species specificity of the blood

The objective of the present study was to evaluate sensitivity and specificity of the HemDirect method on test-plates (Seratec) for detecting human hemoglobin (HHb). These characteristics were compared with those of other widely used methods designed for the detection of blood traces, viz. thin layer chromatography, hemotest, spectrofluorimetry, and identification of blood species specificity (by countercurrent immunoelectrophoresis in agar and on the acetate-cellulose film). It was shown that the HemDirect test is highly specific and far more sensitive than other techniques used for the same purpose in the practical work. It can be recommended as the method of choice for the detection of blood microtraces.     

The value of the Duquénois test for cannabis--a survey.

The Duquénois test is employed throughout the world as part of the identification procedure for cannabis. A survey of results published for more than 400 herbally derived and botanical materials originating from some 270 different plant species is presented. A survey of results from some 200 organic compounds is also presented. The data show that the D-L modification is the most specific. There is no published report of an obviously botanical material, apart from cannabis, that gives a positive D-L test. Apart from cannabinoids, some resorcinol derivatives give a positive test. All of the materials are easily distinguished from cannabis by TLC.     

重新界定《中华人民共和国海商法》的调整对象——以主体-关系规范模式为视角

《中华人民共和国海商法》（简称《海商法》）调整对象的准确界定是该法修改成功的基础之一。《海商法》第1条关于＂调整海上运输关系、船舶关系＂的规定,被认为是《海商法》调整对象的规范表述,但该表述存在概念指称不清、外延过于宽泛或不确定、主体缺失与规定错位及调整对象与调整目标之间缺乏合理的对应关系等问题。关于《海商法》调整对象的学理解释观点纷呈,但均过于表面化,未能反思立法不当问题。通过对《海商法》调整对象界定依据的分析,指出应当依据实践诉求、空间要素、主体法律地位及体系性诉求,将《海商法》调整对象界定为平等主体间的法律地位及因船舶运输、船舶停泊、船舶归属与利用而产生的社会关系及与之有关的社会关系。     

A more sensitive modification of the zinc test for seminal traces suitable for stable test paper strips.

A more sensitive modification of the zinc test for semen has been developed, which can be used either as a solution or incorporated into test papers. The latter retain their sensitivity for at least 3 months.     

Antigenic differentiation of blood stains by the Lewis system in practical forensic medical expert evaluation

A modification of quantitative absorption and absorption elution tests with blood stain washing before the absorption phase is presented. Due to washing, the effect of carrier object on anti-Le(a) and anti-Le(b) sera is decreased and the sensitivity of the method is increased. Additional adsorption of the sera and elution into test erythrocytes treated with protease C is suggested for increasing the number of standard sera fit for the absorption-elution test. A new technology for preparing anti-Le(a) and anti-Le(b) immunoreagents is described.     

A method for group identification of hair infected with Microsporum

A method for identifying the group appurtenance of biological objects from subjects suffering from various diseases is developed. The method can be used in examination of putrefactive objects (blood, secretions, hair, etc.) and in cases when the group appurtenance cannot be identified by other methods.     

Use of a pig model to demonstrate vulnerability of major neck vessels to inflicted trauma from common household items

Commonly available items including a ball point pen, a plastic knife, a broken wine bottle, and a broken wine glass were used to inflict stab and incised wounds to the necks of 3 previously euthanized Large White pigs. With relative ease, these items could be inserted into the necks of the pigs next to the jugular veins and carotid arteries. Despite precautions against the carrying of metal objects such as knives and nail files on board domestic and international flights, objects are still available within aircraft cabins that could be used to inflict serious and potentially life-threatening injuries. If airport and aircraft security measures are to be consistently applied, then consideration should be given to removing items such as glass bottles and glass drinking vessels. However, given the results of a relatively uncomplicated modification of a plastic knife, it may not be possible to remove all dangerous objects from aircraft. Security systems may therefore need to focus on measures such as increased surveillance of passenger behavior, rather than on attempting to eliminate every object that may serve as a potential weapon.     

生命科技背景下的人体与民法上的物

随着生命科技的发展,传统理论上属于人身权客体的人体的一些部分可以脱离人体,进入到物的领域。捐献的器官、血液、精子、卵子、胚胎、切除的病变组织、细胞等用于生命科技的人体生物材料等来自于人体的特殊的物,在特定条件下可以成为物权的客体,而此类物权的内容与行使不同于一般物权,要受到一定的限制。     

Sensitivity of various study technics in blood stains

Quadratic pieces of fleece measuring 16 mm2 were soaked with 10 different blood-samples in the dilution steps of 1:1, 1:10, 1:100, 1:1000, respectively, and were tested in blood group typing and identification tests of forensic serology. The above spezified dilutions correspond with 5 microliters, 0.5 microliter, 0.05 microliter and 0.005 microliter of blood, respectively. The detection limit of the microspectrometric test for blood was the dilution 1:10, of the porphyrine test a dilution above 1:100, whereas the preliminary test for blood (peroxidase) succeeded always up to a dilution of 1:1000 and the species determination by the radial immunodiffusion test in agar gels succeeded in most cases op to a dilution of 1:1000. The detection limit of the anti-human globulin inhibition test was between the dilution steps 1:10 and 1:100 when non-titrated and undiluted anti-human globulin serum was used. Gc- and ABO-grouping were possible up to a dilution of 1:100 and were thus the most sensitive grouping systems. Phenotyping of the enzyme-systems and the Gm/Km-system usually required stains with considerably higher blood concentrations i.e. stains of undiluted blood.     

Developmental validation of a novel lateral flow strip test for rapid identification of human blood (Rapid Stain Identification™-Blood)

Human blood is the body fluid most commonly encountered at crime scenes, and blood detection may aid investigators in reconstructing what occurred during a crime. In addition, blood detection can help determine which items of evidence should be processed for DNA-STR testing. Unfortunately, many common substances can cause red-brown stains that resemble blood. Furthermore, many current human blood detection methods are presumptive and prone to false positive results. Here, the developmental validation of a new blood identification test, Rapid Stain Identification™-Blood (RSID™-Blood), is described. RSID™-Blood utilizes two anti-glycophorin A (red blood cell membrane specific protein) monoclonal antibodies in a lateral flow strip test format to detect human blood. We present evidence demonstrating that this test is accurate, reproducible, easy to use, and highly specific for human blood. Importantly, RSID™-Blood does not cross-react with ferret, skunk, or primate blood and exhibits no high-dose hook effect. Also, we describe studies on the sensitivity, body fluid specificity, and species specificity of RSID™-Blood. In addition, we show that the test can detect blood from a variety of forensic exhibits prior to processing for DNA-STR analysis. In conclusion, we suggest that RSID™-Blood is effective and useful for the detection of human blood on forensic exhibits, and offers improved blood detection when compared to other currently used methods.     

Validation of presumptive tests for non-human blood using Kastle Meyer and Hemastix reagents

Kastle Meyer and Hemastix reagents are presumptive tests commonly used in forensic casework for the detection of blood, and their suitability has been reviewed in numerous publications. However, studies to date have focused on the validation of these tests on human blood alone, and no published work has looked at the sensitivity, specificity and effect on DNA analysis when using these reagents to presumptively test for animal blood. The aim of this study was to validate the two reagents for use with animal blood, and compare their performance in order to choose the best test based on the circumstances in wildlife crime investigation.The sensitivity, specificity, stability and robustness of the methods were assessed by experiments with dilutions of animal blood (from 1:4 to 1:65536) using direct and indirect (rub) tests, potential interfering substances, blood sources from different species and aged blood. The effects of the two reagents on subsequent DNA analysis were also investigated.During the direct tests, Kastle Meyer showed a higher sensitivity, detecting blood down to a dilution of 1:16,384, one order of magnitude lower than Hemastix. However during the rub test, Hemastix showed a higher sensitivity, detecting blood down to a dilution of 1:64 on porous materials while Kastle Meyer was positive only down to a dilution of 1:16. Moreover, when using the same swab for presumptive testing and DNA extraction, Hemastix testing allowed amplification of a sufficient amount of DNA for species identification at its limit of sensitivity on porous materials (1:64) while Kastle Meyer inhibited most amplification of DNA at its less sensitive limit of 1:16 dilution. On the other hand, Hemastix showed a much lower specificity, producing false positive results when exposed to tomato, potato, rust, avian uric acid, bleach and sink rot, while Kastle Meyer only produced a faint positive reaction from potato. Both tests performed equally well detecting fresh blood of different animal species. The stability test gave comparable results among the tests except for aged fish blood stains, where the Kastle Meyer test performed poorly.Owing to its ease of use, higher sensitivity, and lack of interference with downstream DNA analysis, and despite its reduced specificity compared to Kastle Meyer, the Hemastix method is more appropriate for use in wildlife crime investigations. Positive results would always be confirmed with DNA analysis and the low interference of the reagent will allow the use of a single swab for presumptive testing and DNA sampling.     

An extremely sensitive species-specific ARMs PCR test for the presence of tiger bone DNA

The survival of the tiger (Panthera tigris) is seriously threatened by poaching to provide raw materials for Traditional Chinese Medicines (TCMs). Most highly prized are the tiger's bones, which are used in combination with other animal and plant derivatives in pills and plasters for the treatment of rheumatism and other ailments. Hundreds of patent remedies have been produced which claim to contain tiger bone, but proof of its presence is needed, if legislation prohibiting the trade in endangered species is to be enforced. A highly sensitive tiger-specific real-time PCR assay has been developed to address this problem. Using primers specific to the tiger mitochondrial cytochrome b gene, successful amplification has been reliably achieved from blood, hair and bone as well as from a range of TCMs spiked with 0.5% tiger bone. Although capable of detecting fewer than 10 substrate molecules, the seven varieties of TCM pills and plasters tested showed no detectable trace of tiger DNA before spiking. Furthermore, sequencing several "tiger bone" fragments seized from TCM shops has shown that they actually originated from cattle and pigs. The potential effects of traditional bone preparation methods, evidence that much lower concentrations are used than alleged on TCM packaging, and substitution of bones from other species all suggest a low likelihood of detecting tiger DNA in patent medicines. Despite this, the basic methods have been thoroughly proven and can be readily applied to derivatives from other CITES protected species providing a rapid and highly sensitive forensic test for species of origin. Potential applications to the monitoring of wild populations are demonstrated by the successful identification of shed hairs and faecal samples.     

Forensic and chemical determination of methane in cadaveric samples

Conditions were elaborated for the determination of methane, through gas chromatography, in blood and the lung by using a 2% methanol solution as an internal standard and with the below gas-chromatography column being applied: dimensions--200 x 0.3 cm, 15% di-2-ethylhexyl sebacate on Dinochrome II (0.16 x 0.21 mm) at 110 0 degree C. The detector is of the flame-ionization type. The normal content of methane was determined for blood and for the lung--0.122 +/- 0.0074 microgram/ml and 0.20 +/- 0.04 microgram/g, respectively. Biological samples from cadavers of other persons who died due to trauma were suggested for use as controls. The method was used to examine expertise objects, blood and the lung from the cadavers of peoples who died in fire.     

The establishment of the presence of sperm in stains by an agar gel electrophoretic method]

Comparative investigation of seminal stains and other secretions from human body and blood as well as objects of plant origin by gel electrophoresis using alkaline values of pH buffer solutions and subsequent enzymography for acid phosphatase made it possible to develop optimal conditions for detection of seminal presence in stains. Good preservation of seminal acid phosphatase in stains during several years is shown.     

Modified cobalt thiocyanate presumptive color test for ketamine hydrochloride

A new presumptive color test for ketamine hydrochloride is reported. The test is a modification of the cobalt thiocyanate test currently used for cocaine and involves basifying samples rather than acidifying them. The two-step procedure for liquids and three-step procedure for powdered samples are straightforward, definitive, and utilize reagents commonly used in forensic drug analysis. The test works on ketamine hydrochloride in both powder and liquid form and has a sensitivity of c. 1.25 mg. Performing the test with numerous other controlled substances and related chemicals demonstrates the test to be highly selective.     

An extremely sensitive species-specific ARMS PCR test for the presence of tiger bone DNA

The survival of the tiger (Panthera tigris) is seriously threatened by poaching to provide raw materials for traditional Chinese medicines (TCMs). Most highly prized are the tiger's bones, which are used in combination with other animal and plant derivatives in pills and plasters for the treatment of rheumatism and other ailments. Hundreds of patent remedies have been produced which claim to contain tiger bone, but proof of its presence is needed if legislation prohibiting the trade in endangered species is to be enforced.A highly sensitive tiger-specific real-time PCR assay has been developed to address this problem. Using primers specific to the tiger mitochondrial cytochrome b gene, successful amplification has been reliably achieved from blood, hair and bone as well as from a range of TCMs spiked with 0.5% tiger bone. Although capable of detecting fewer than 10 substrate molecules, the seven varieties of TCM pills and plasters tested showed no detectable trace of tiger DNA before spiking. Furthermore, sequencing several "tiger bone" fragments seized from TCM shops has shown that they actually originated from cattle and pigs. The potential effects of traditional bone preparation methods, evidence that much lower concentrations are used than alleged on TCM packaging, and substitution of bones from other species all suggest a low likelihood of detecting tiger DNA in patent medicines. Despite this, the basic methods have been thoroughly proven and can be readily applied to derivatives from other Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) protected species, providing a rapid and highly sensitive forensic test for species of origin. Potential applications to the monitoring of wild populations are demonstrated by the successful identification of shed hairs and faecal samples.     

A novel method for ABO genotyping using a DNA chip

ABO genotyping is often performed to identify the blood type of decomposed samples, which is difficult to be determined by a serological test. In this study, we developed a simple method for ABO genotyping using a DNA chip. In this method, polymerase chain reaction-amplified and fluorescent-labeled fragments in the ABO gene and primate-specific D17Z1 were hybridized with DNA probes on a chip designed to detect single nucleotide polymorphisms (SNPs) in the ABO gene and part of the D17Z1 sequence. Using blood samples from 42 volunteers and 10 animal species, we investigated whether the chip could be used to detect SNPs in the ABO gene and the D17Z1 sequence. This method was then applied to various forensic samples, and it was confirmed that this method was suitable for the simultaneous analyses of ABO genotyping and species identification. This method fulfills the recent need for the development of rapid and convenient methods for criminal investigations.     

A microthermal resistance method for determining the species specificity of protein

Microthermal method was used for objective registration of the course of precipitation reaction in determination of species specificity of human and animal protein. This method allows one to carry out species identification using blood protein dilution with 0.9% solution of natrium chloride in correlation from 1:100 up to 1:100,000. Method is simple in use, it can be operated on the apparatus of home production. Analysis with two control tests may be performed within 10 min. The results of reaction are successfully registered as thermistograms subjoined to the expert conclusion.     

微波衍生化和GC/MS/MS技术分析血、尿中的吗啡

目的建立了血、尿等生物检材中海洛因代谢产物吗啡的定性分析方法。方法以丙酸酐为衍生化剂,采用微波衍生化技术结合GC/MS/MS进行分析。结果当CID电压为0.9V时,衍生化物的母离子与子离子碎片信息丰富,碎片为m/e268、324、342。结论此方法科学、准确,灵敏度高,能满足吸食海洛因类、吗啡类毒品人员的生物检材的检验要求。