1,2,4‐Triazine‐Based Chromogenic Reagents for the Detection of Microtraces of Various Metals Left on Human Skin*

Abstract: This article extends the application of 1,2,4‐triazine‐based chromogenic reagents to the detection of nonferrous metal traces left on contact with canvas and human skin. The possibility of detection of iron traces resulting from contact with objects made of stainless steel was investigated as well. Additionally, the ability of triazines to form chromatic complexes with Zn²⁺, Ni²⁺, Co²⁺, Cu²⁺, Cr³⁺, and Al³⁺ ions was studied spectrophotometrically. Molar absorption coefficients, ranging from 8.8 to 29.9 × 10³/M/cm, provide high sensitivity of 1,2,4‐triazines toward nonferrous ions, thus, enabling the detection at concentrations as low as a few μM. The method was sensitive enough to detect traces resulting from a 1‐min contact with a stainless steel made object, which is commonly considered as a corrosion‐resistant material. The amounts of metal ions transferred to the skin after a 2‐min contact with objects made of brass, zinc, and copper were sufficient to develop chromatic imprints.     

2,4,6-Tri(2′-pyridyl)-1,3,5-triazine for determination of iron(II), Iron(III), and total iron contents on human palms

When iron-based tools, such as knives or guns, are held, traces of iron can transfer to the skin. However, no previous studies have been published regarding the effect of the elapsed contact time on the transfer of iron species with different valences to the palm. Compared with 3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine (PDT), 2,4,6-tri(2′-pyridyl)-1,3,5-triazine (TPTZ) was found to exhibit a higher sensitivity to iron(II) spectrophotometrically. This work employed 2,4,6-tri(2′-pyridyl)-1,3,5-triazine (TPTZ) and UV spectrophotometry to determine the amounts of iron(II), iron(III), and total iron transferred to human palms from iron tools. It was found that the palmar moisture level was an important factor in determining the amounts of total iron, including iron(II), transferred to the palm. For identical contact times, the amounts of total iron transferred to the palm was proportional to the palmar moisture, and the difference between the maximum and minimum amounts was 12 μg per hand. However, the amounts of iron(II) transferred to the palm gradually decreased over time for low palmar moisture levels, but steadily increased over time for high palmar moisture levels. Additionally, for average levels of palm moisture, the amounts of iron(II) and iron(III) transferred to the palm gradually decreased and increased, respectively, with longer contact times. Notably, this research could serve as a theoretical basis and guide for the detection of trace iron species with different valences on human palms for criminal investigations.     

Can mRNA markers distinguish traces generated by different types of contact?

The presence of contact traces on objects can be caused by different activities such as personal use, firm gripping or casual touching. As these activities are of distinct duration and intensity, we hypothesised that cell material corresponding to different epidermal depth may be deposited. Epidermal layers have distinguishable biomarker profiles [1], [2] that are formed by cell proliferation, cell differentiation and cell death. Thus, we examined whether mRNAs corresponding to different epidermal layers can distinguish the different types of contact. From an initial set of 14 candidates, six epidermal markers showed sufficient expression in contact traces to be included in an mRNA multiplex assay that carried body fluid markers as well [3]. Using a set of 30 user and 30 grip traces left by identical donors on identical substrates, we determined that none of these epidermal mRNAs could specify contact type. When the corresponding DNA data were included in the analyses no relation with contact type could be established either.     

Trace evidence dynamics of cocaine on banknotes: A comparison study of paper and polymer banknotes

It is well established that a large proportion of paper banknotes in circulation contain traces of cocaine. Being able to discriminate between the innocent transfer of illicit drug particles acquired through everyday interactions with surfaces such as banknotes, as opposed to transfer resulting from criminal activities can provide valuable intelligence that can inform an investigation. With many countries adopting polymer banknotes as legal tender, it is important to consider the transfer of cocaine from these surfaces as well as the retention of these particulates on polymer banknotes for evaluative interpretation in crime reconstruction. This comparison study assessed three contact variables (force, time, and rotation) on the transfer of cocaine particulates from paper and polymer banknotes onto a human skin proxy. The persistence of cocaine particulates was assessed through a realistic scenario which mimicked a cash transaction. Quantifiable amounts of cocaine were transferred onto the human skin proxy across all of the contacts assessed, with a greater transfer observed with contacts involving polymer banknotes and those contacts which involved rotation. Following extensive handling, cocaine persisted on both banknote types, with paper banknotes retaining larger amounts of cocaine than polymer banknotes. These findings show that cocaine can persist on both paper and polymer banknotes for extended periods of time following handling and is therefore available for transfer. This transfer then readily occurs, even when contact is brief and involves relatively small forces. A key distinction between the banknote types was that cocaine particulates are more likely to transfer from polymer banknotes due to the lower retention rate of particulates on this surface. Such insights can aid in evaluating the relevance of illicit drug particles identified on items or persons of interest in crime reconstruction approaches.     

Significance of skin metallization in the diagnosis of electrocution

The differentiation of minute current marks from the faint traces of burns may offer some difficulties. Marcinkowski and Wojciechowski (1973) treated the skin of corpses with the same metal objects either heated to a high temperature or exposed to 250 V of alternating current, and determined (by an electrographic method) that metallization appeared only after applying the electric current.The continuation of these observations is linked with the actual experimental studies of Pankowski. By treating the skin of corpses with alternating and direct current of 10, 50, 100 and 250 V for 0.3 sec, 1 sec, 30 sec and 1 min passing through a radioactive electrode of ⁶⁰Co, he has shown that the radioactivity of the skin at the site of electrode contact increases with the elevation of the voltage and its duration. In the case of direct current the rise was 630 – 54000-fold at the site of the positive electrode.Using electrodes of copper, aluminium and iron (not radioactive) it has also been shown (by an electrographic method) that metallization intensifies under the same conditions of time and voltage.Metallization could be detected even when no current marks on the skin were evident. Electrography appears to be extremely useful in the detection of metallization. No metallization was detected at the site of the negative electrode (as refers to direct current).     

Recovery of Latent Fingerprints and DNA on Human Skin*

Abstract: The project “Latent Fingerprints and DNA on Human Skin” was the first systematic research in Europe dealing with detection of fingerprints and DNA left by offenders on the skin of corpses. One thousand samples gave results that allow general statements on the materials and methods used. The tests were carried out according to a uniform trial structure. Fingerprints were deposited by natural donors on corpses. The latent fingerprints were treated with magnetic powder or black fingerprint powder. Afterward, they were lifted with silicone casting material (Isomark^®) or gelatine foil. All lifts were swabbed to recover DNA. It was possible to visualize comparable and identifiable fingerprints on the skin of corpses (16%). In the same categories, magnetic powder (18.4%) yielded better results than black fingerprint powder (13.6%). The number of comparable and identifiable fingerprints decreased on the lifts (12.7%). Isomark^® (14.9%) was the better lifting material in comparison with gelatine foil (10.1%). In one‐third of the samples, DNA could be extracted from the powdered and lifted latents. Black fingerprint powder delivered the better result with a rate of 2.2% for full DNA profiles and profiles useful for exclusion in comparison with 1.8% for the magnetic powder traces. Isomark^® (3.1%) yielded better results than gelatine foil (0.6%).     

The detection of metallic residues in skin stab wounds by means of SEM-EDS: A pilot study

The morphological analysis of stab wounds may often not be accurate enough to link it with the type of wounding weapon, but a further evaluation may be performed with the search for metallic residues left during the contact between the instrument and the skin. In this study, Scanning Electron Microscopy-Energy Dispersive Spectroscopy (SEM-EDS) was applied to the study of cadaveric stab wounds performed with kitchen knives composed of iron, chromium and nickel, in order to verify the presence of metallic residues on the wound's edge. Two groups of 10 corpses were selected: group A, including victims of stab wounds and a control group B (died of natural causes). Samplings were performed on the lesions and in intact areas of group A, whereas in group B sampling were performed in non-exposed intact skin. Samples were then analysed with optical microscopy and SEM-EDS. In group A, optical microscopic analysis showed the presence of vital haemorrhagic infiltration, while SEM-EDS showed evidence of microscopic metal traces, isolated or clustered, consisting of iron, chromium and nickel. Moreover, in two cases organic residues of calcium and phosphate were detected, as a probable sign of bone lesion. Control samples (group A in intact areas and group B), were negative for the search of exogenous material to optical microscopy and SEM-EDS. The results show the utility and possible application of the SEM-EDS in theidentification of metallic residues from sharp weapons on the skin.     

From laboratory to the street: Capturing witness memory using the Self‐Administered Interview

The Self‐Administered Interview (SAI^©) is a tool designed to elicit a comprehensive initial account from witnesses at the scene of an incident or shortly thereafter to inoculate against the loss of information associated with delayed interview. Drawing on the principles of the Cognitive Interview (CI), the SAI^© provides witnesses with a series of instructions and retrieval cues to support recall. Requesting that witnesses complete an SAI^© not only serves to preserve and protect memory but also enables officers to prioritize the allocation of policing resources during the critical early stages of an investigation. The current review traces the development of the SAI^© from a series of laboratory studies through to field trials and integrates our findings with theoretical accounts of human memory. We present new data from trials of the tool in the field and consider future avenues for research and further development of the SAI^©.     

Could Secondary DNA Transfer Falsely Place Someone at the Scene of a Crime?,

The occurrence of secondary DNA transfer has been previously established. However, the transfer of DNA through an intermediary has not been revisited with more sensitive current technologies implemented to increase the likelihood of obtaining results from low‐template/low‐quality samples. This study evaluated whether this increased sensitivity could lead to the detection of interpretable secondary DNA transfer profiles. After two minutes of hand to hand contact, participants immediately handled assigned knives. Swabbings of the knives with detectable amounts of DNA were amplified with the Identifiler^® Plus Amplification Kit and injected on a 3130xl. DNA typing results indicated that secondary DNA transfer was detected in 85% of the samples. In five samples, the secondary contributor was either the only contributor or the major contributor identified despite never coming into direct contact with the knife. This study demonstrates the risk of assuming that DNA recovered from an object resulted from direct contact.     

Detection of Contact Traces of Powdery Substances

The current practice in securing the contact traces of chemical substances taken from clothes belonging to a person suspected of manual handling explosives is focused on pockets and cuffs. The outerwear worn by people who had contact with fluorescent powders that simulate explosives and drugs was the subject of this study. Clothes were first exposed to the test substance for a period of time and then analyzed by fluorescence methods to determine the location of the highest quantity of traces. The results obtained from the study confirm that the areas with the highest concentration of powdery traces are different from those suggested by current forensic practice. They appear to be promising for a more efficient identification of the suspects involved in illegal manufacturing of drugs of abuse or explosives. Moreover, they may be helpful for developing the methodology for handling the evidence material in the forensic clothing examination process.     

A Study on the Estimation of Postmortem Interval Based on Environmental Temperature and Concentrations of Substance in Vitreous Humor

A method to determine postmortem interval (PMI) based on environmental temperature and the concentrations of vitreous humor (VH) molecules were explored. Rabbit carcasses were placed in a chamber at 5, 15, 25, or 35°C, and 80–100 μL of VH was collected with the double‐eye alternating micro‐sampling method every 12 h. A Roche DPPI biochemical analyzer was used to measure the concentrations of six substances in VH samples. The interpolation function model and mixed‐effect model were employed for data fitting to establish equations for PMI estimation. The concentrations of K⁺, P, Mg²⁺, creatinine (CRE), and urea nitrogen (UN) exhibited an upward trend with increasing PMI in all temperature groups, while the concentration of Ca²⁺ showed a downward trend. Validation results using K⁺ and Mg²⁺ ions revealed that the mixed‐effect model provided a better estimation than the interpolation function model using the data from our experiment. However, both models were able to estimate PMI using temperature and VH molecule concentrations.     

Analyzing Forensic Evidence Based on Density with Magnetic Levitation

This paper describes a method for determining the density of contact trace objects with magnetic levitation (MagLev). MagLev measurements accurately determine the density (±0.0002 g/cm³) of a diamagnetic object and are compatible with objects that are nonuniform in shape and size. The MagLev device (composed of two permanent magnets with like poles facing) and the method described provide a means of accurately determining the density of trace objects. This method is inexpensive, rapid, and verifiable and provides numerical values—independent of the specific apparatus or analyst—that correspond to the absolute density of the sample that may be entered into a searchable database. We discuss the feasibility of MagLev as a possible means of characterizing forensic‐related evidence and demonstrate the ability of MagLev to (i) determine the density of samples of glitter and gunpowder, (ii) separate glitter particles of different densities, and (iii) determine the density of a glitter sample that was removed from a complex sample matrix.     

Identification of human skin from a tissue fragment by detection of squamous cell carcinoma-related antigen using an enzyme immunoassay

A new method of identifying human skin from a tissue fragment by detection of squamous cell carcinoma-related (SCC) antigen, using an enzyme immunoassay, was developed. When an extract was prepared from 0.1 g human skin homogenized with 1 ml of phosphate buffered saline, this method was able to detect SCC antigen in extracts diluted 10²-fold. There was no difference in the detection limit between individuals. Species specificity was good, and there was no cross reaction observed with skins from animals. Our method could also discriminate between skin and other organs or tissues, except for esophagus and lung. A practical case to which this method was applied is presented.     

Development of a Protein‐based Human Identification Capability from a Single Hair

Shed human hair (lacking root nuclear DNA) frequently contributes important information to forensic investigations involving human identification. Detection of genetic variation observed in amino acid sequences of hair proteins provides a new suite of identity markers that augment microscopic hair analysis and mitochondrial DNA sequencing. In this study, a new method that completely dissolves single hairs using a combination of heat, ultrasonication, and surfactants was developed. Dissolved proteins were digested and genetically variant peptide (GVP) profiles were obtained for single hairs (25 mm) via high‐resolution nanoflow liquid chromatography‐based mass spectrometry and a novel exome‐driven bioinformatic approach. Overall, 6519 unique peptides were identified and a total of 57 GVPs were confirmed. Random match probabilities ranged between 2.6 × 10^?2 and 6.0 × 10^?9. The new bioinformatic strategy and ability to analyze GVPs in forensically relevant samples sizes demonstrate applicability of this approach to distinguish individuals in forensic contexts.     

水、油污中无电流斑电击死大鼠皮肤和心肌的病变

目的探讨水中、油污中无电流斑电击死法医学鉴定的病理形态学依据。方法 SD大鼠28只,分为水中、油污中无电流斑电击死各1组,典型电流斑组、正常对照组、死后电击组、死后水中、油污中电击各1组共7组。采用肉眼、光镜及投射电镜观察水中、油污中无电流斑电击死大鼠皮肤和心肌组织病理学改变,并与其它各组进行比较。结果采用肉眼观察,生前水中、油污中无电流斑电击死大鼠皮肤未见明显电流斑。普通光镜观察,可见电击中心部位表皮变性坏死、脱落,表皮细胞变薄、致密,表皮细胞或/和毛囊、汗腺、皮脂腺发生极性化改变。电镜观察,透明层和角质层分离脱落,基底细胞肿胀、细胞器减少、核固缩,汗腺导管上皮肿胀,棘细胞中粗面内质网扩张融合成泡状,线粒体肿胀空泡化;但光镜与电镜的变化与生前电击死比较不明显、典型。而死后电击组皮肤则无明显病理学改变。实验各组大鼠心肌的改变与皮肤改变类同。结论采用光镜和投透射电镜观察在潮湿环境中电击死的组织病理学改变,可为无电流斑电击死提供依据。     

应用扫描电镜能谱仪检测电击伤皮肤表面的金属元素——附一例案例报告

作者采用已知化学组成的导体对家兔进行触电试验。取受损部位的皮肤作常规处理,用带能谱的扫描电镜作金属离子测定,仪器型号为 JXA-840-LinK-860。对检出的金属元素,进行了定性和半定量分析,并观察了元素的分布。作者在一例人体电击伤皮肤活检标本上检出铁元素,而对照皮肤则无,从而认定是含铁的导体与皮肤接触。电击伤与热烫伤的超微结构变化能区别。     

An innovative transfer DNA experimental design and qPCR assay to identify primary,secondary, and tertiary DNA transfer

“Touch DNA” is a form of trace DNA that is presumed to be deposited when an individual touches something and leaves behind DNA-containing skin cells, sweat, or other fluids. While touch DNA is often the result of direct contact (i.e., primary transfer), it can also be indirectly transferred between surfaces or individuals (e.g., secondary or tertiary transfer). Even experts cannot distinguish between different types of transfer and do not fully understand which variables affect direct versus indirect transfer or how often each type of transfer occurs. In this study, we utilize an innovative protocol that combines a paired male and female transfer DNA experimental design with an Amelogenin qPCR assay to generate data on primary, secondary, and tertiary DNA transfer. We report frequencies of indirect DNA transfer and also investigate the potential effects of participant age, self-identified ethnicity, and skin conditions on DNA transfer. Out of 22 experimental trials, we detected primary transfer (male + female) in 71% of trials, secondary DNA transfer in 50% of trials, and tertiary DNA transfer in 27% of trials. No significant associations were found between primary DNA transfer and age, self-identified ancestry, or skin conditions, however, all individuals with sloughing skin conditions demonstrated primary DNA transfer and we suggest this variable be explored in larger samples. These results contribute to a better understanding of the conditions under which secondary and tertiary DNA transfer occurs and can be used to propose realistic DNA transfer scenarios in court cases.     

Corrosion behaviour of four handguns in aqueous environments: Corrosion product characterization and effects on estimating the time since deposition

When a firearm has been disposed of in a body of water and becomes corroded, its appearance is altered and determining a time-since-immersion may be of import to the investigation. Therefore, in this study, the corrosion and mass loss of four handgun slides over a period of 180 days were examined. Solid-state characterization of the metals and their corrosion products via SEM/EDX and powder X-ray Diffraction (pXRD) was performed. The pXRDs were analyzed against the NIST Powder Diffraction Database to determine the crystalline phases. Filings from the SS416 standard, Llama and Ruger handgun slide predominantly consisted of iron alloys. After 180-days in solution, pXRD indicated that the adherent corrosion products consisted of 1) γ-FeOOH and 2) iron oxide (Fe₃O₄ or Fe₂O₃). Additionally, pXRD analysis indicated that the adherent corrosion products of the SS416 standard also consisted of CrO₃. Metal filings from the Raven and Jennings handgun slides were a mixture of iron–nickel–zinc and EDX and pXRD analyses of the corrosion products, when submersed in deionized water, indicated that the products consisted of: 1) γ-FeOOH, 2) iron oxide (Fe₃O₄ or Fe₂O₃), and 3) ZnFe₂O₄ or ZnO; where the Jennings adherent rust contained ZnFe₂O₄ and the Raven adherent rust contained ZnO. Further, pXRD of the corrosion products from these alloys, when submersed in 25 PSU (Practical Salinity Unit) solution, indicated that the products consisted of: 1) ZnO, 2) Zn(OH)₂, 3) α-Ni(OH)₂, and 4) NaCl.The data thus indicated that both metal composition and the presence of chloride ions had significant impacts on rates and products of corrosion and suggest that the presence of Cl^? changes not only the rate of corrosion, but also the corroding species itself. While mechanisms and rates of the chloride driven corrosion processes offer explanations as to the different oxides and hydroxides observed between immersion conditions, they do not offer an explanation for the differences observed between handguns. Therefore, utilizing a general approach where surface area coverage of corrosion products is the sole consideration is not sufficient to determine time-since-immersion. Attempts to determine a time-since-immersion would require a priori knowledge of the mechanism of corrosion for a given metal mixture within a specified environment. The results described herein give indications as to the possible corrosion mechanism driving the process in high and low Cl^? environments and show the necessity of including the metal composition, rust composition and ion concentration in any models that attempt to elucidate the time-since-immersion of handguns for forensic applications.     

Screening Test for Shed Skin Cells by Measuring the Ratio of Human DNA to Staphylococcus epidermidis DNA

A novel screening method for shed skin cells by detecting Staphylococcus epidermidis (S. epidermidis), which is a resident bacterium on skin, was developed. Staphylococcus epidermidis was detected using real‐time PCR. Staphylococcus epidermidis was detected in all 20 human skin surface samples. Although not present in blood and urine samples, S. epidermidis was detected in 6 of 20 saliva samples, and 5 of 18 semen samples. The ratio of human DNA to S. epidermidisDNA was significantly smaller in human skin surface samples than in saliva and semen samples in which S. epidermidis was detected. Therefore, although skin cells could not be identified by detecting only S. epidermidis, they could be distinguished by measuring the S. epidermidis to human DNA ratio. This method could be applied to casework touch samples, which suggests that it is useful for screening whether skin cells and human DNA are present on potential evidentiary touch samples.     

New York State TrueAllele® Casework Validation Study

DNA evidence can pose interpretation challenges, particularly with low‐level or mixed samples. It would be desirable to make full use of the quantitative data, consider every genotype possibility, and objectively produce accurate and reproducible DNA match results. Probabilistic genotype computing is designed to achieve these goals. This validation study assessed TrueAllele^® probabilistic computer interpretation on 368 evidence items in 41 test cases and compared the results with human review of the same data. Whenever there was a human result, the computer's genotype was concordant. Further, the computer produced a match statistic on 81 mixture items (for 87 inferred matching genotypes) in the test cases, while human review reported a statistic on 25 of these items (30.9%). Using match statistics to quantify information, probabilistic genotyping was shown to be sensitive, specific, and reproducible. These results demonstrate that objective probabilistic genotyping of biological evidence can reliably preserve DNA identification information.