Fluorescence Imaging of Posterior Spiracles from Second and Third Instars of Forensically Important Chrysomya rufifacies (Diptera: Calliphoridae),

Entomological protocols for aging blowfly (Diptera: Calliphoridae) larvae to estimate the time of colonization (TOC) are commonly used to assist in death investigations. While the methodologies for analyzing fly larvae differ, most rely on light microscopy, genetic analysis, or, more rarely, electron microscopy. This pilot study sought to improve resolution of larval stage in the forensically important blowfly Chrysomya rufifacies using high‐content fluorescence microscopy and biochemical measures of developmental marker proteins. We established fixation and mounting protocols, defined a set of measurable morphometric criteria and captured developmental transitions of 2nd instar to 3rd instar using both fluorescence microscopy and anti‐ecdysone receptor Western blot analysis. The data show that these instars can be distinguished on the basis of robust, nonbleaching, autofluorescence of larval posterior spiracles. High‐content imaging techniques using confocal microscopy, combined with morphometric and biochemical techniques, may therefore aid forensic entomologists in estimating TOC.     

Trace Elemental Analysis of Titanium Dioxide Pigments and Automotive White Paint Fragments for Forensic Examination Using High‐Energy Synchrotron Radiation X‐Ray Fluorescence Spectrometry*

Abstract: High‐energy synchrotron radiation x‐ray fluorescence spectrometry (SR‐XRF) utilizing 116 keV x‐rays was used to characterize titanium dioxide pigments (rutile) and automotive white paint fragments for forensic examination. The technique allowed analysis of K lines of 9 trace elements in 18 titanium dioxide pigments (rutile), and 10 trace elements in finish coat layers of seven automotive white paint fragments. High‐field strength elements (HFSE) were found to strongly reflect the origin of the titanium dioxide (TiO₂) pigments, and could be used as effective parameters for discrimination and classification of the pigments and paint fragments. A pairwise comparison of the finish coat layers of seven automotive white paint fragments was performed. The trace elements in the finish coat layers detected by the high‐energy SR‐XRF were especially effective for identification. By introducing the trace element information of primer and electrocoat layers, all the automotive white paint fragments could be discriminated by this technique.     

Detection of pretreated fingerprint fluorescence using an LED-based excitation system

Abstract:  Optimization of a light emitting diode (LED)-based excitation system for the detection of pretreated fingerprint fluorescence is described. Fluorescent ridges can usually be excited by irradiation with forensic light sources such as xenon arc lamps or quartz-halogen lamps with high-power output and suitable filters. However, they are too expensive for many crime laboratories in smaller organizations. We concentrated on LEDs which have advantages over conventional light sources in that they are simpler and of lower cost, but the power output and quality of each individual LED unit is not sufficient for the detection of weak fluorescent ridges. To resolve this subject, blue and green LED arrays composed of ninety LED units were adopted and suitable low pass filters for them were designed. An experimental system, consisting of blue and green LED arrays with the suitable low pass filters for illumination, high pass filters for viewing, a digital camera and a computer, was tested. The fluorescent images of cyanoacrylate ester fumed/rhodamine 6G stained fingerprint on white polyethylene sheet and weak fluorescent ridges of ninhydrin/indium chloride treated fingerprint on white paper were successfully detected and photographed. It was shown that the improvement of LED beam in intensity and quality can compensate the disadvantages, resulting in well-contrasted images.     

Comprehensive evaluation of pericardial biochemical markers in death investigation

Pericardial fluid (PCF) is a well-preserved cadaveric material in cases without structural damage. The present study investigated fundamental serum components of PCF, including total proteins (TP), albumin (Alb), urea nitrogen (UN), creatinine (Cr), uric acid (UA), glucose (Glu), sodium (Na), potassium (K), chloride (Cl), calcium (Ca) and magnesium (Mg) in PCF with regard to the postmortem and survival periods, and cause of death in serial medicolegal autopsy cases (n = 288) with intact pericardial and cardiac structures within 48 h postmortem. The amount of PCF (mostly 5–25 ml) showed no survival or postmortem time dependence, or difference among the causes of death. For all cases, there were moderate postmortem decreases of Na and Cl, and increases of K and Mg, which were insufficient for application to estimate the time since death; however, characteristic findings with regard to the cause of death were detected in cases of hours-long survival, especially in ‘functional causes of death’: higher Alb, UN, Cr, UA, K and Mg, and lower Na, Cl and Ca in intoxication; lower TP, Alb, Cr, K and Mg, and higher Na and Cl in hypothermia (cold exposure); higher Alb, UN, Cr, UA and Mg, and lower Glu and Ca in hyperthermia (heatstroke). These observations suggest the usefulness of pericardial biomarkers for investigating the cause and process of death to reinforce pathological and toxicological findings.     

Unexpected death due to intestinal obstruction by a duplication cyst in an infant

An 11-month-old female infant died at home after being diagnosed as having gastroenteritis. Autopsy examination revealed a distended distal ileum filled with a large amount of watery contents, and a 3 cm x 3 cm x 4 cm cyst at the ileocecal part of the cecum that was histologically consistent with a duplication cyst. The cause of death was determined to be acute dehydration due to intestinal obstruction caused by a duplication cyst. This case indicates that intestinal obstruction by a duplication cyst should be recognized as a cause of unexpected death in infants.     

Detection of diatom in formalin-fixed tissue by proteinase K digestion

The diatoms detection has been proposed to be useful in the diagnosis of drowning. Enzymatic digestion of unfixed lung tissues and other organs with proteinase K is widely employed to detect diatoms. Handling unfixed organs or blood from the bodies with some infectious diseases could prove to be dangerous. In this study, we examined the application of enzymatic digestion for diatom detection to formalin-fixed lung obtained at autopsy. Furthermore, we assessed the effect of hydrogen peroxide on the contamination of the lung specimen with foreign bodies inhaled in the course of drowning, smoking, or air pollution. Formalin-fixed lung was heated in 0.01 M Tris–HCl buffer (pH 7.5) containing sodium dodecyl sulfate (SDS) (tissue lysis-buffer), with or without glycine. Thereafter, the lung was subjected to enzymatic digestion with proteinase K. A part of formalin-fixed or unfixed samples digested with proteinase K were incubated with hydrogen peroxide at 80 °C for 6 h or 12 h, while the residues were processed without incubation. Formalin-fixed samples heated in tissue lysis-buffer with glycine could be digested with proteinase K; further, the number and proportion of diatoms detected in both formalin-fixed and unfixed samples were observed to be similar. The results suggest that enzymatic detection of diatoms can be applied to formalin-fixed organs by heating the samples in glycine-containing tissue lysis-buffer. As the use of formalin-fixed tissue for diatom detection can decrease risk of contamination by pathogenic organisms during the course of enzymatic digestion, the method presented in this study would be beneficial, to some extent, to individuals performing diatom analysis. Moreover, our results suggest that archival organs stored in formalin solution could be available in diatom detection over a long time-period following autopsies. Clearer image of diatoms was observed in the specimen incubated with hydrogen peroxide for 6 h, in which inhaled foreign bodies were discolored, than those not subjected to incubation. Therefore, incubation of sample digested with hydrogen peroxide in the limited time would be helpful for quantitative and qualitative diatom analysis.     

Lower limits of detection of synchrotron radiation high-energy X-ray fluorescence spectrometry and its possibility for the forensic application for discrimination of glass fragments

Synchrotron radiation high-energy X-ray fluorescence (SR-XRF) analysis utilizing 75.5keV X-ray radiation from beam-line BL37XU at Super Photon Ring 8GeV (SPring-8), a third-generation synchrotron facility, was found to have advantages for forensic discrimination of glass samples. The lower limits of detection (LLD) for calibration curves were at the picogram level for Ba, Ce, and Sm and at the 10pg level for Sr, Zr, Sn, and Hf. The spectrum of NIST SRM 612 glass reference material demonstrated K-line peaks of 31 elements including rare-earth elements, and the relative standard deviations (R.S.D.) of all the measured elements except Ca were less than 9.7%. Fragments of collected sheet glass were used as samples for investigating the application of this technique to forensic analysis. Several trace elements such as Pb, Rb, Sr, Zr, La, Ce, and Hf were detected in the spectra of the samples, and these elements could be used as indexes to characterize the glass samples. But the "lower limits of detection (LLD)" of each element were not examined enough. In this report, these limits by synchrotron radiation X-ray spectrometry were clarified. By these results, this technique should provide an effective approach to the nondestructive discrimination of small glass fragments in the field of forensic science.     

The stem family and labour markets: Reflections on households and firms in Japan's economic development

This paper examines a view that the traditional stem family system was one of the preconditions for Japan's modern economic development, focusing on labour markets and skill formation practices. The paper begins with a brief look at the Japanese stem family household formation rules. Then, exploration is made, first, on the self-employed, the largest sector of the early modern economy; second, the merchant house and its employment patterns as an origin of present day large corporations' employment system and skill formation and human capital management practices; and third, workshop industries, which formed middle and lower layers of the manufacturing sector in the period of industrialisation. Finally, women's marriage behaviour is examined in relation to labour markets, especially changes in real wages. All this is an attempt to go some way towards a better understanding of the ways in which the family economy and corporate firms worked in economic development, rather than to suggest an alternative hypothesis on the relationship between family and household factors and subsequent economic development.     

Preliminary study of post mortem identification using lip prints

Identification using lip prints was first performed in the 1950s and was the subject of much research in the 1960s and 70s, leading to the acceptance of this technique as evidence in the criminal justice system. Previous research has focused on identifying lip print types or on methods of obtaining hidden lip prints left at the crime scene. The present study aimed to clarify characteristics of lip prints from cadavers with various causes of death (including drowning and hanging) and to determine the effects of fixation on post mortem changes in lip impressions.