Evaluation of four digestive methods for extracting diatoms

Objective

To compare the merit of four digestive methods (nitric acid plus hydrogen peroxide, proteinase K, nitric acid in Disorganization Can and Soluene-350) for extracting diatoms in order to choose the best digestive method for the diagnosis of drowning.

Methods

Liver, kidney and bone marrow of rabbits were minced and then digested by four digestive methods separately with the following indices compared: (1) time demanded for complete digestion; (2) degree of digestion for different tissues; (3) the reclaiming ratio of diatoms; (4) the degree of digestive destruction to diatoms.

Results

For sufficiently digesting the same tissue, the demanded times for the different methods ranked from the longest to the shortest were as follows: Soluene-350, proteinase K, nitric acid plus hydrogen peroxide, nitric acid in Disorganization Can. Nitric acid in Disorganization Can method and nitric acid plus hydrogen peroxide method digested the tissues more thoroughly than proteinase K, than Soluene-350 methods. For Cyclotella and Cybella, proteinase K method reclaimed most diatoms and nitric acid plus hydrogen peroxide method reclaimed less, while nitric acid in Disorganization Can and Soluene-350 methods reclaimed the least. For Navicula, the majority of diatoms could be extracted using proteinase K method, but only a few diatoms with other three methods. Under scanning electron microscopy (SEM), the structure of diatoms remained almost perfect after digestion with proteinase K, but destroyed to some extent with other three methods.

Conclusion

This study demonstrated that different diatoms (in fresh or sea water) have different resistance to different digestive reagents. As far as the reliability and applicability of the diatom test is concerned, proteinase K method is of the best choice, nitric acid plus hydrogen peroxide can be its substitute. Soluene-350 cannot be used for extracting sea water diatoms.     

大蒜素对卵巢癌细胞株HO8910生长的抑制作用

目的：观察大蒜素对卵巢癌细胞株HO8910的抑制作用。方法；用不同时间不同剂量大蒜素作用于HO8910，通过肿瘤细胞药敏试验。观察肿瘤细胞存活率，并通过透射电镜观察一定剂量的大蒜素作用于HO8910一定时间后，肿瘤细胞形态的改变。结果：50-100μg/ml大蒜素可显抑制肿瘤细胞的生长。且在作用2h时即出现显的抑制，随着作用时间的延长，抑制作用愈明显，72h害到高峰，透射电镜结果显示：50μg/ml的大蒜素作用HO891024h。细胞肿胀。细胞膜破碎，胞浆脱落。细胞核裸露；小剂量（25μg/ml)则HO8910呈现细胞凋亡样改变；细胞胞浆呈空泡，染色质边集，核仁固缩，胞质呈片状脱落。结论：大蒜素可显抑制HO8910的生长，且有明显的时间及剂量效应，大剂量的大蒜素对HO8910细胞呈直接杀伤作用。而小剂量则诱导细胞凋亡。     

扫描电镜/能谱观察溺死和死后入水的法医学应用研究

目的探讨扫描电镜/能谱分析在溺死的法医学鉴定中的作用。方法实验选用20例已知生前入水尸体,11例已知的死后入水尸体和2例陆地死亡尸体。在每个肺叶各取3个切面,每个切面用扫描电镜取样台胶面与肺叶的切面充分接触,另在落水地点取水样一份,进行对比。扫描电镜取样台胶在切面上充分接触后封闭取材盒,真空干燥后,直接在扫描电镜下进行观察。结果20例生前入水尸体在肺组织中均检出异物颗粒,并且与对照水样中检出的异物颗粒相吻合;而死后入水的尸体和在陆地上死亡的尸体肺组织中均未检出明显的异物颗粒,3者比较P<0.0001。在肺组织支气管切面中,生前入水的20例中有15例检出异物颗粒,在边缘肺切面及中间切面中分别有13例检出异物颗粒,统计学无明显差异。结论本实验建立的肺脏内异物颗粒取材方法准确可靠实用。对综合判断案件的性质,判断溺死尸体入水地点的具有积极意义。较好地解决了实验室检验中的污染问题。腐败尸体对检测结果无影响。     

家兔烧死后气管、肺及心肌的病理形态学观察

目的探讨烧死的病理形态学依据。方法建立家兔烧死动物模型,用光镜、透射及扫描电镜观察10只生前烧死家兔气管、肺及心肌的病变;并与死后焚尸及非烧死的正常组(各5只)对照。结果透射电镜下,气管粘膜上皮完全脱落,成堆红细胞和纤维蛋白成分粘附在平滑肌表面。肺Ⅰ型上皮细胞结构基本完整连续,线粒体嵴有断裂、缺失;Ⅱ型肺泡上皮细胞微绒毛消失,多数细胞线粒体、板层小体、高尔基体未见异常,少数Ⅱ型肺泡上皮细胞质膜破损、染色质浓集、胞质内容丢失。心肌严重变性坏死,细胞膜破坏,线粒体极度肿胀,肌丝溶解并出现收缩带坏死。扫描电镜下,气管粘膜上皮有完整纤毛,但上皮大片脱落,少数区域纤毛上皮稀疏排列,有脱落柱状上皮附着在纤毛上;肺间皮细胞结构基本完整,微绒毛数量不一致;心内膜的内皮细胞大部脱落,暴露内皮下结缔组织,心肌纤维呈扭曲排列。死后焚尸及正常组对照组未见上述变化。结论烧死兔的光镜和电镜下气管、肺及心肌的病理形态学变化,能为生前烧死与死后焚尸的鉴别提供依据。     

Microscopic Residues of Bone from Dissolving Human Remains in Acids

Dissolving bodies is a current method of disposing of human remains and has been practiced throughout the years. During the last decade in the Netherlands, two cases have emerged in which human remains were treated with acid. In the first case, the remains of a cremated body were treated with hydrofluoric acid. In the second case, two complete bodies were dissolved in a mixture of hydrochloric and sulfuric acid. In both cases, a great variety of evidence was collected at the scene of crime, part of which was embedded in resin, polished, and investigated using SEM/EDX. Apart from macroscopic findings like residual bone and artificial teeth, in both cases, distinct microscopic residues of bone were found as follows: (partly) digested bone, thin‐walled structures, and recrystallized calcium phosphate. Although some may believe it is possible to dissolve a body in acid completely, at least some of these microscopic residues will always be found.     

Analysis and discrimination of electrical tapes: part II. Backings

The backings of 90 black electrical tapes were analyzed to evaluate the chemical components of these films, the ability of individual techniques to discriminate samples, and the ability of the techniques combined to distinguish samples. The techniques utilized and their respective discrimination results were stereomicroscopy and physical measurements, to include observation of surface features of the backing, width, and thickness measurements (c. 64%); Fourier transform infrared spectroscopy (FTIR) using a microscope accessory (c. 83%); pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS; c. 81%); and scanning electron microscopy/energy-dispersive spectroscopy (SEM/EDS; c. 87%). Ninety-four percent of the backings were discriminated through this combination of analytical methods. Finally, evaluating these results in conjunction with previously published data on the analysis of the adhesives from the same set of electrical tapes provided an overall discrimination of nearly 96%.     

The identification of two unusual types of homemade ammunition

Illegal homemade ammunition is commonly used by criminals to commit crimes in Taiwan. Two unusual types of homemade ammunition that most closely resembling genuine ammunition are studied here. Their genuine counterparts are studied as the control samples for the purpose of comparison. Unfired ammunition is disassembled, and the morphological, dimensional, and compositional features of the bullet and cartridge case are examined. Statistical tests are employed to distinguish the dimensional differences between homemade and genuine ammunitions. Manufacturing marks on head stamps of the cartridge case are carefully examined. Compositional features of propellant powders, primer mixtures, and gunshot residues are also analyzed. The results reveal that the morphological, dimensional, and compositional features of major parts of the ammunition can be employed to differentiate homemade cartridges from genuine ones. Among these features, tool marks on the head stamps left by the bunter can be used to trace the origin of ammunition.     

电流斑的环境扫描电镜X射线能谱分析

目的利用环境扫描电镜-X射线能谱仪(environmental scanning electron microscope and energydispersive X-ray microanalyser,ESEM-EDX)研究电流斑微观形态特征及元素构成,以期为电流斑及电击死的鉴定提供更准确、客观的依据。方法用ESEM-EDX检测5例电流斑的扫描电镜形态、金属颗粒的形态及其元素组成。结果电流斑及周围皮肤可见电穿孔、金属熔珠,熔珠主要由铁、铜、铝等常见金属元素和金、钛、钡等人体少见的金属元素构成。结论 ESEM-EDX检测可为电击死的鉴定提供客观的依据。     

Comparison of GSR Composition Occurring at Different Locations Around the Firing Position

Abstract: Variations in gunshot residue (GSR) compositions are used in the reconstruction of shooting incidents. In this study, GSR samples taken from seven different locations around and in the firearm were collected and analyzed using scanning electron microscopy/energy dispersive X‐ray analysis. Four different types of ammunition were applied. Very low correlations were found when different ammunition were used. This clearly shows that it is possible to differentiate between ammunition types. When the same ammunition was used, high correlations were found between samples taken from external positions (such as hands of shooter, bullet‐entrance holes) but poor correlation was found between internal samples (such as firearm barrel, cartridge case) and external samples. A high degree of association was found between samples that simulated victim and shooter. These findings clearly demonstrate that GSR comparison studies are meaningful but care needs to be taken when choosing suitable exhibits. External samples (such as hands of shooter, bullet‐entrance holes) are more suitable candidates than internal samples (barrel of the firearm, cartridge case).     

Analysis of Gunshot Residue and Associated Materials—A Review

Abstract: A comprehensive review of the scientific literature on gunshot residue (GSR) is presented. Aspects of both inorganic and organic GSR are discussed, from formation and distribution, to sample collection, preparation, and analysis using a variety of techniques. The interpretation of GSR results is also considered including issues surrounding the contamination, distribution, and transfer of GSR. Potential problems with ulterior sources of GSR like particles have been reported in the literature. For example, particles from environmental and occupational sources have been highlighted as exhibiting similar chemical and morphological characteristics to GSR. These findings are put into context with regard to interpreting samples. A move toward a “case by case” approach is argued to be more preferable to a “formal” classification system where possible. The analysis of both inorganic and organic compositions of residue samples as well as morphological considerations is considered to be a more ideal approach to GSR analysis, whereever practicable.