Forensic chemical detection of furadan

Isolation of furadan from biological material by ethyl acetate is considered. Column chromatography with silicone gel L 100/160micro was used to purify extractions from biological objects. A technique was elaborated for identification and quantitation of furadan in cadavers. It is possible to use this technique in forensic expert examination.     

大鼠口服敌敌畏和呋喃丹急性中毒后血液胆碱酯酶体外自动复能的动态观察

作者测定了中毒鼠血胆碱酯酶活性在体外室温下的动态变化,观察研究胆碱酯酶自动复能的特点。结果表明中毒鼠血胆碱酯酶活性体外自动复能在72小时内达最大值;敌敌畏中毒鼠血胆碱酯酶自动复能较呋喃丹组幅度大而下降也快;至第17天,与对照组相比,敌敌畏中毒鼠血胆碱酯酯活性仍有显著性差异。根据实验结果就中毒鼠血胆碱酯酶自动复能的法医学意义作了讨论。     

大鼠口服敌敌畏和呋喃丹急性中毒后乙酰胆碱酯酶组织化学的研究

作者通过大鼠口服敌敌畏和呋喃丹急性中毒实验,利用乙酰胆碱酯酶组织化学技术,研究中毒鼠肋间肌、空肠和大脑乙酰胆碱酯酶活性受抑制和自动复能情况、血液的体外抑制效应以及正常大鼠在室温和冷藏条件下空肠和肋间肌乙酰胆碱酯酶活性的保存时间,探讨利用乙酰胆碱酯酶组织化学方法诊断胆碱酯酶抑制剂中毒的法医学意义,并对大鼠敌敌畏和呋喃丹中毒后上述观察指标的不同特点作了分析比较。     

检测浮游生物叶绿素相关基因诊断溺死的实验研究

目的评估浮游生物叶绿素相关基因检测用于溺死诊断的价值。方法将18只大白兔随机分为溺死组（n=10）、死后抛尸组（n=6）和对照组（n=2）,各组分别取心血及肺、肝、肾、脑组织,分离浮游生物并提取其DNA,用PCR技术检测叶绿素相关基因EG（EG1和EG2）及SK（SK1和SK2）。同时用硝酸消化法检验肺和肝组织中的硅藻。结果溺死组心血及肺、肝、肾、脑组织中EG1分别检出9、10、9、7和8例阳性,EG2分别检出8、10、7、5和7例阳性;死后抛尸组仅在心血和肺组织中各检出1例EG1阳性;对照组各组织均未检出EG1和EG2。SK1、SK2除在溺死组心血、肺和肾有少数检出外（≤2例）,在其他组未检出扩增产物。硝酸消化法从溺死组肺、肝组织中分别有9例及3例检出硅藻,死后抛尸组仅在1例肺组织中检出。结论浮游生物叶绿素相关基因EG用于溺死诊断的阳性检出率要高于硝酸消化法,在溺死诊断中具有较高的应用价值。     

DYF155S1基因座的法医学应用研究

目的探讨Y染色体小卫星DYF155S1基因座在法医学中的应用价值。方法采用荧光MVR-PCR和Amp-FLP技术。结果同一个体不同组织所获得的分型结果一致;本方法能作出正确分型的最低基因组DNA量为0.3 ng;在女性成份为男性成份100倍的混合DNA样本(总DNA量多少为110 ng)中能正确检出DYF155S1基因型;调查130个父系家庭的男性成员样品(减数分裂336次),总共发现DYF155S1基因座有3次突变,突变率为0.9%。对强奸案混合斑的分析,可直接检出DYF155S1基因型。但是,本方法只能区分哺乳类雄性动物与非哺乳类动物。结论采用荧光MVR-PCR和Amp-FLP方法分析Y染色体小卫星DYF155S1基因座,方法可靠,灵敏度高,对混合斑和微量检材中男性DNA的分析具有较高的应用价值。     

A simple automated instrument for DNA extraction in forensic casework

The Qiagen BioRobot EZ1 is a small, rapid, and reliable automated DNA extraction instrument capable of extracting DNA from up to six samples in as few as 20 min using magnetic bead technology. The San Diego Police Department Crime Laboratory has validated the BioRobot EZ1 for the DNA extraction of evidence and reference samples in forensic casework. The BioRobot EZ1 was evaluated for use on a variety of different evidence sample types including blood, saliva, and semen evidence. The performance of the BioRobot EZ1 with regard to DNA recovery and potential cross-contamination was also assessed. DNA yields obtained with the BioRobot EZ1 were comparable to those from organic extraction. The BioRobot EZ1 was effective at removing PCR inhibitors, which often co-purify with DNA in organic extractions. The incorporation of the BioRobot EZ1 into forensic casework has streamlined the DNA analysis process by reducing the need for labor-intensive phenol-chloroform extractions.     

Extraction of diquat with 1-butanol from biological materials

Diquat can be extracted with 1-butanol from high pH solution in the presence of several moderate reductants. The red colored reduced compound of diquat in water turns to a purple compound in 1-butanol. The absorption of the purple compound is 0.105 at 383 nm and 0.119 at 520 nm in 1 microgram diquat/ml 1-butanol. The latter value is a little higher than that of the red compound at 495 nm in water. The purple compound is much more stable than the red compound in water. More than 80% of 10 ppm diquat added can be extracted from serum, blood, tissues, urine and some drinks. The extraction with 1-butanol is useful for concentration of diquat contained in large volume. The lower limit of detection is 0.1 microgram/ml 1-butanol. Paraquat is insoluble in 1-butanol under the same condition. Therefore, this method is applicable for the determination of diquat when paraquat is also contained in the solution.     

Rapid isolation with Sep-Pak C18 cartridges and capillary gas chromatography of triazine herbicides in human body fluids.

A simple and rapid method, for the isolation of eight triazine herbicides from human serum and urine, using Sep-Pak C18 cartridges is presented. After mixing with distilled water, serum and urine samples containing the herbicides, were loaded on Sep-Pak C18 cartridges and eluted with either chloroform only or chloroform/methanol (9:1). The herbicides were detected by capillary gas chromatography with both flame ionization detection (FID) and nitrogen-phosphorus detection (NPD). Separation of eight triazine herbicides from each other and from impurities was generally satisfactory with the use of a non-polar DB-1 capillary column. Recovery of most compounds was excellent for both chloroform and chloroform/methanol (9:1) as elution solvents. Backgrounds were cleaner and evaporation time was shorter for the chloroform only than for the chloroform/methanol (9:1). The NPD gave sensitivity more than 10-20 times higher than that of FID.     

Allelic frequencies for the HLA-DQA1, D1S80, HUMTHO1, HUMTPOX, HUMCSF1PO and HUMVWA loci in Cantabria (middle north Spain)

Allele frequencies for six DNA polymorphisms have been studied in a population sample from Cantabria (middle north Spain) using the polymerase chain reaction. The HLA-DQA1 locus was analyzed by the reverse dot-blot technique and the other five by polyacrylamide gel electrophoresis followed by silver staining. Six alleles were found for HLA-DQA1. 15 alleles for D1S80, 6 alleles for HUMTHO1 and HUMCSF1PO, 7 for HUMTPOX and 8 alleles for HUMVWA. The 21 repeat allele in HUMVWA had not previously been reported in a Spanish population. The genotype distributions met Hardy-Weinberg expectations for all the systems and some statistical parameters of forensic interest were calculated. Comparisons with other populations revealed significant differences for HLA-DQA1, HUMVWA and HUMTHO1, with interracial differences being more pronounced than between Spanish populations. The HUMVWA system showed the highest forensic efficiency of the six polymorphisms studied.     

Separation and structural characterization of the synthetic cannabinoids JWH-412 and 1-[(5-fluoropentyl)-1H-indol-3yl]-(4-methylnaphthalen-1-yl)methanone using GC-MS, NMR analysis and a flash chromatography system

The 'herbal highs' market continues to boom. The added synthetic cannabinoids are often exchanged for another one with a high frequency to stay at least one step ahead of legal restrictions. While most of these substances were synthesized for pharmaceutical purposes and have been described in the scientific literature before, others originate from clandestine laboratories supplying this lucrative market. In this paper, the identification and structure elucidation of two synthetic cannabinoids is reported. The first compound, 1-[(5-fluoropentyl)-1H-indol-3yl]-(4-methylnaphthalen-1-yl)methanone, was found along with AM-2201 in a 'herbal mixture' obtained via the Internet. For isolation of the substance from the mixture, a newly developed flash chromatography method was used providing an inexpensive and fast way to gain pure reference substances from 'Spice' products for the timely development or enhancement of analytical methods in the forensic field. The second substance, 4-fluoronaphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-412) was seized by German authorities as microcrystalline powder, making it very likely that it will be found in 'herbal mixtures' soon.     

GC polymorphism detected in human urine by isoelectric focusing and immunoblotting

Genetic polymorphism of GC (vitamin D-binding protein) in human urine was revealed by isoelectric focusing and immunoblotting on thin-layer polyacrylamide gels containing 2 M urea. Urine samples from 530 unrelated Japanese from the Fukui district, being only 1-2 ml of original urine, were examined, and correct GC typing was achieved by comparison with the results of direct grouping using plasma. Six common and twelve rare phenotypes were observed. The frequencies of the genes were 0.473 for GC*1F, 0.241 for GC*1S, 0.254 for GC*2, and 0.032 for the total of six rare alleles.     

Analysis of DNA in minute volumes of blood from stains and crusts

As the first step, the locus D1S80 was amplified by the polymerase chain reaction technique from genomic DNA extracted from artificial bloodstains and crusts with different amount of blood (32 microl, 16 microl, 8 microl, 4 microl, 2 microl, and 1 microl). In all samples of bloodstains and crusts, identification by DNA analysis was possible. As the second step, the locus HLA-DQA1 was amplified from genomic DNA extracted from diluted blood samples (640, 320, 160, 80, 40, 20, 10, and 5 leukocytes). DNA amplification was possible in diluted blood samples with at least 10 leukocytes. Considering the conditions in which the present study was carried out, it was possible to conclude that 1 microl of bloodstains or crusts was enough for identification. It was also concluded that five leukocytes are not enough material to render consistent DNA identification.     

冠心病猝死心肌mcl-1蛋白检测及其意义

目的观察冠心病猝死(SCD)心肌mcl-1蛋白产物,探讨其免疫组化检测及其对SCD诊断的意义。方法运用免疫组织化学SABC法,对46例SCD和40例非猝死心肌(有冠心病和无冠心病)中mcl-1蛋白产物进行检测和观察,并比较其差异。结果(1)自症状出现至死亡,时间超过30min的SCD(36例),其心肌组织均出现mcl-1蛋白阳性染色;(2)自症状出现至死亡,时间短于30min的SCD(10例),其心肌组织mcl-1蛋白呈弱阳性染色;(3)冠心病非猝死样本(20例),4例心肌出现微弱的mcl-1蛋白阳性染色,无冠心病非猝死样本(20例)几乎没有出现阳性染色。结论心肌mcl-1蛋白的免疫组化检测可诊断自症状出现至死亡时间超过30min的SCD。     

Expression of adhesion molecules in skin wounds: diagnostic value in legal medicine

The adhesion molecules identified in recent years can help improve the diagnosis of the wound age, especially of injuries with a short survival time. This is also indicative of the vitality of the wounds. The material investigated in the study originated from 465 skin wounds. The samples were taken from human autopsy material, during the surgical treatment of wounds (excision) of patients and from experimental incised wounds of mice. To judge the age of skin wounds the endothelial adhesion molecules were detected in paraffin sections after autoclaving and using the ABC technique. Human skin wounds: strong positive staining was observed of ICAM-1 1.5 h at the earliest and 3.5 days at the latest, for the P-selectin 3 min at the earliest and 7 h at the latest, for the E-selectin 1 h at the earliest and 17 days at the latest and for VCAM-1 3 h at the earliest and 3.5 days at the latest after the time of injury. The L-selectin was expressed constitutively. Mice skin wounds: strong positive immunohistochemical reactions were found as a rule earlier than in human skin wounds. The detection of an increased expression of ICAM-1, VCAM-1 and P- and E-selectins can improve the wound age assessment in injuries with short survival times.     

Medicare program; changes to the hospital inpatient prospective payment systems and fiscal year 1996 rates; corrections--HCFA. Correction to final rule

In the September 1, 1995, issue of the Federal Register (60 FR 45778), we published a final rule with comment period revising the Medicare hospital inpatient prospective payment systems for operating costs and capital-related costs to implement necessary changes arising from our continuing experience with the system. In the addendum to that final rule with comment period, we announced the prospective payment rates for Medicare hospital inpatient services for operating costs and capital-related costs applicable to discharges occurring on or after October 1, 1995, and set forth update factors for the rate-of-increase limits for hospitals and hospital units excluded from the prospective payment systems. This document corrects errors made in that document.     

Medicare program; changes to the hospital inpatient prospective payment systems and fiscal year 1995 rates; correction--HCFA. Final rule; correction

In the September 1, 1994 issue of the Federal Register (59 FR 45330), we published a final rule with comment period revising the Medicare hospital inpatient prospective payment systems for operating costs and capital-related costs to implement necessary changes arising from our continuing experience with the system. In the addendum to that final rule with comment period, we announced the prospective payment rates for Medicare hospital inpatient services for operating costs and capital-related costs applicable to discharges occurring on or after October 1, 1994, and set forth update factors for the rate-of-increase limits for hospitals and hospital units excluded from the prospective payment systems. This notice corrects errors made in that document.     

急性心肌缺血猝死心肌中血红素加氧酶-1的表达

目的观察血红素加氧酶-1（HO-1）在急性心肌缺血猝死心肌细胞胞浆中的表达情况。方法应用免疫组织化学技术（SP法）,对27例可疑早期心肌缺血猝死（早期梗死组）、10例心肌梗死猝死（心肌梗死组）、11例冠心病非心性猝死（对照组1）以及10例正常心肌（对照组2）中HO-1的表达进行观察分析。结果早期梗死组与心肌梗死组在心肌细胞胞浆中HO-1均呈强阳性表达,对照组1和对照组2心肌细胞胞浆中HO-1弱阳性表达或无表达。早期梗死组和心肌梗死组与对照组1和对照组2之间心肌细胞内HO-1表达的差异有显著性意义（P（0.01）;镜下各组心肌细胞胞浆内HO-1表达的阳性面积率及平均光密度差别明显。结论 HO-1可以作为急性心肌缺血所致心性猝死的一个诊断指标。     

血液中64种常见有机氯菊酯类农药的快速筛查

目的建立人体血浆中64种有机氯菊酯类农药多残留的气相色谱（GC）快速筛查分析方法。方法空白人体静脉抗凝血用乙腈沉淀蛋白,乙酸乙酯∶环己烷（v/v,3∶1）进行液液萃取净化,使用HP-5色谱柱,采用气相色谱进行定性、定量分析。对方法进行优化并进行方法学评价。结果 64种农药在0.001~0.1μg/mL范围内线性关系均良好,相关系数在0.990 1~0.999 9之间。检出限在0.001~0.15μg/mL范围内,方法定量限在0.001~0.5μg/mL之间,回收率总体于80%~110%之间,相对标准偏差小于15%,方法检出限总体于0.01μg/mL以下,日内精密度在1.5%~11.5%之间,日间精密度在2.9%~13.9%之间。结论本文建立的人体血浆的农药多残留快速筛查测定法,符合农药残留分析方法的要求,可在相关研究和实践中选用。     

成人肾脏ABO组织血型1-4型糖链H物质的分布

目的观察1型H、2型H及3/4型H糖链在成人肾组织中的分布及其与分泌状态的关系。方法 应用抗ABO抗体及3种糖链特异的单克隆抗H抗体的免疫组织化学方法,检查分泌型与非分泌型个体肾组织中相应抗原物质的分布。结果 在分泌型和非分泌型人的肾远曲小管均表达2型H和3/4型H物质,1型H和3/4型H物质只在分泌型人肾集合管表达,在非分泌型人中不表达。另外集合管的2型H物质的表达与分泌状态无关。结论 人肾组织有ABH物质的表达,不同肾组织细胞表达的H物质结构差异与AB0型分泌状态有关。     

Physical Characterization and Recovery of Corroded Fingerprint Impressions from Postblast Copper Pipe Bomb Fragments

Pipe bombs made from 1 mm thick copper pipe were detonated with a low explosive power powder. Analysis of the physical characteristics of fragments revealed that the copper had undergone work hardening with an increased Vickers Hardness of 107HV1 compared with 80HV1 for unexploded copper pipe. Mean plastic strain prior to fracture was calculated at 0.28 showing evidence of both plastic deformation and wall thinning. An examination of the external surface showed microfractures running parallel with the length of the pipe at approximately 100 μm intervals and 1–2 μm in width. Many larger fragments had folded “inside out” making the original outside surface inaccessible and difficult to fold back through work hardening. A visual examination for fingerprint corrosion revealed ridge details on several fragments that were enhanced by selective digital mapping of colors reflected from the surface of the copper. One of these fingerprints was identified partially to the original donor.