Kiyoshi Saigusa

A histological study on the mechanism of epidermal nuclear elongation in electrical and burn injuries.

Abstract Epidermal nuclear elongation is one of the most important signs for the diagnosis of electrical injury. In this study, we investigated the mechanism responsible for this phenomenon by comparing the findings from burn injuries and those from contusions. Electrical and burn injuries were made in the dorsal skin of rats using energy ranging from 100 to 790 joules for electrical injury, and 170–690 joules for burn injury. Contusions were also made by compressing the skin with a vice. In electrical and burn injuries, the dermis under the epidermal elongated nuclei was homogeneous and without empty spaces between collagen bundles and the number of dermal fibroblasts per 0.01 mm2 below the damaged epidermis decreased significantly (P < 0.05). The incidence of this change correlated with the depth of denatured dermal collagen fibres and in both types of injuries, dermal cells had no nuclear antigenicity for ubiquitin. The width of the injured epidermis with nuclear elongation decreased significantly (P < 0.05) and the elongated nuclei were parallel to the basal membrane. In electrical injury however, nuclear elongation occurred more frequently near the external root sheath. Nuclear elongation of fibroblasts and external root sheath cells was also found, but those of sebaceous gland cells were not detected. Epidermal elongated nuclei were also found in contusions. The evidence strongly suggests that epidermal nuclear elongation in electrical and burn injuries is due to dermal expansion by heat.

Wound age estimation by simultaneous detection of 9 cytokines in human dermal wounds with a multiplex bead-based immunoassay: An estimative method using outsourced examinations

Wound age estimation for human dermal wounds was performed based on quantification of interleukin 1beta (IL 1beta), IL 5, IL 7, IL 12 p70, IL 13, IL 17, granulocyte colony-stimulating factor (G-CSF), monocyte chemoattractant protein 1 (MCP 1), and macrophage inflammatory protein 1beta (MIP 1beta). IL 5, IL 12 p 70, IL 13, and IL 17 increased from the early phase, MCP 1 exclusively in the middle phase, and IL 1beta, G-CSF, and MIP 1beta from the middle phase to the late phase. IL 7 decreased from the early phase. Among the cytokines analyzed in the present study, MCP 1 was the most plentiful cytokine. In addition, an outsourced examination, which could be available to any forensic institute, was performed in two cases for confirmative purposes. Many factors have been proposed as markers for dermal wound age estimation, but the set of cytokines selected for the outsourced examination in the present study wound be useful in daily forensic practice.

A case of acute gasoline intoxication at the scene of washing a petrol tank.

We encountered a case of acute gasoline intoxication at the scene of washing the inner wall of a petrol tank. The decedent was a 50-year-old male, who was the supervisor. Two young workers wearing mask respirators began to wash the inner wall of the gasoline tank under poor ventilation. About 1 h later, because one of the workers lost consciousness, the supervisor entered the tank, without a mask respirator, to rescue the worker. However, the supervisor immediately fainted, and died 26 h after the accident. In the autopsy, blisters were observed on the skin of the face, neck, anterior chest, upper extremities, and back. The heart contained hemolyzed blood. Histologically, hemorrhagic pulmonary edema, pneumonia, and proximal tubular necrosis were found. In the toxicological analyzes, toluene, xylene, and trimethylbenzene were detected in the blood, brain, and gastric contents. Toluene concentrations in the blood and brain were 0.3 mug/ml and 3.7 mug/g, respectively. Since pathological findings were consistent with the reported findings concerning gasoline intoxication, and constituents of gasoline were in the body, death was attributed to acute gasoline intoxication. It was considered that sufficient ventilation and proper use of a mask respirator were essential for this kind of work.

A study on mRNA expressions of interleukin 10 during fracture healing for wound age determination

We investigated mRNA expressions of interleukin 10 (IL-10) during fracture healing for wound age determination. Macroscopically, calluses were formed at 144 h post-injury, and thickening of the fracture edge was observed at 240 h post-injury. Slight infiltration of neutrophils and macrophages was detected at 3 h post-injury, and moderate infiltration of T and B lymphocytes was detected at 72 h post-injury. In addition, osteoblasts proliferated at 24 h post-injury. IL-10 mRNA increased immediately after the intervention and peaked at 24 h, and the expressions were detected in osteocytes before and after injury, and in osteoblasts at 3 h post-injury. Although the intensity was stable in osteocytes, that in osteoblasts was strongest at 24 h post-injury. Among inflammatory cells, the expressions were exclusively detected in macrophages. These results indicated that examinations of macroscopic and microscopic changes in fracture edges, and mRNA expressions of IL-10, would be informative for fracture healing and dating.

Practical applications of molecular biological species identification of forensically important flies.

Species identification of the early colonizers on a corpse, such as Calliphoridae and Sarcophagidae, provides useful information for estimating the postmortem interval (PMI). However, the morphology of adult flies and nymphs is very similar among species of the same genus, and for many forensic scientists accurate identification is very difficult within their routine work. We present a simple and time-saving method for identification of forensically important flies by comparing sequenced 304 bp of mitochondrial DNA cytochrome oxidase gene subunit I (mtDNA COI). Eggs and larvae were recovered from the corpse during forensic autopsies and then reared in a growth chamber under controlled conditions. The emerging adults were killed by ethyl acetate and male specimens were observed under a stereomicroscope to identify the species based on the shape of the genitalia. Subsequently, DNA was extracted from identified specimens and sequenced mtDNA COI, resulting in the identification of 8 species of 4 genera of forensically important diptera. Moreover, the molecular biological species identification was useful in forensic practice, and there were cases in which analysis of DNA extracted from puparia supposed to be early colonizers of the corpse provided useful information for estimating the PMI. We also report sequencing of mtDNA COI of the green bottle fly, Lucilia ampullacea, newly collected from a corpse in Iwate prefecture.

The Applicability of Holography in Forensic Identification: A Fusion of the Traditional Optical Technique and Digital Technique

In this study, the applicability of holography in the 3-dimensional recording of forensic objects such as skulls and mandibulae, and the accuracy of the reconstructed 3-D images, were examined. The virtual holographic image, which records the 3-dimensional data of the original object, is visually observed on the other side of the holographic plate, and reproduces the 3-dimensional shape of the object well. Another type of holographic image, the real image, is focused on a frosted glass screen, and cross-sectional images of the object can be observed. When measuring the distances between anatomical reference points using an image-processing software, the average deviations in the holographic images as compared to the actual objects were less than 0.1 mm. Therefore, holography could be useful as a 3-dimensional recording method of forensic objects. Two superimposition systems using holographic images were examined. In the 2D-3D system, the transparent virtual holographic image of an object is directly superimposed onto the digitized photograph of the same object on the LCD monitor. On the other hand, in the video system, the holographic image captured by the CCD camera is superimposed onto the digitized photographic image using a personal computer. We found that the discrepancy between the outlines of the superimposed holographic and photographic dental images using the video system was smaller than that using the 2D-3D system. Holography seemed to perform comparably to the computer graphic system; however, a fusion with the digital technique would expand the utility of holography in superimposition.

Forced oral opening for cadavers with rigor mortis: two approaches for the myotomy on the temporal muscles

Forensic dentistry plays an essential role in personal identification procedures. An adequate interincisal space of cadavers with rigor mortis is required to obtain detailed dental findings. We have developed intraoral and two directional approaches, for myotomy of the temporal muscles. The intraoral approach, in which the temporalis was dissected with scissors inserted via an intraoral incision, was adopted for elderly cadavers, females and emaciated or exhausted bodies, and had a merit of no incision on the face. The two directional approach, in which myotomy was performed with thread-wire saw from behind and with scissors via the intraoral incision, was designed for male muscular youths. Both approaches were effective to obtain a desired degree of an interincisal opening without facial damage.

An application of PCR-single strand conformation polymorphism to MN genotyping.

A PCR-based genotyping of MN blood group system was investigated for DNA samples taken from a population of 409 northern Japanese. DNA fragment (257bp) including exon 2 of glycophorin A (GPA) gene, in which encodes the determinants of MN antigens, was specifically amplified. On the analysis of PCR-single-strand conformation polymorphism (PCR-SSCP) for M alleles, band patterns of M(G) and M(T) were easily discriminated each other. For N alleles, three band patterns were observed, and we tentatively named these alleles as N(1), N(2) and N(V). The N(1) allele appeared predominantly and N(2) had two base substitutions at 1st (C-->A) and 56th (C-->T) in exon 2 of N(1). The other N(V), which was detected from a pair of a mother and her child, possessed a single base substitution at 23rd (A-->G) in intron 2. The allele frequencies of M(G), M(T), N(1) and N(2) were 0.4450, 0.0978, 0.4303 and 0.0269, respectively. The polymorphism information content and the probability of paternity exclusion by this MN genotyping were estimated to be 0.5252 and 0.3219, respectively.

Sudden infant death with anomalous origin of the left coronary artery from the pulmonary artery

Autopsy of a 2-month-old boy revealed an anomalous origin of the left coronary artery from the pulmonary trunk. The endomyocardial fibrosis associated with myocardial ischemia was noted in the left ventricular wall of the enlarged heart. Death was attributed to silent heart failure due to the anomalous left coronary artery. Significant clinical symptoms are generally observed in patients with such anomalies, however, they might cause sudden death in childhood.

An autopsy case of aortitis resulting in sudden death due to a rupture of aneurysm of the aortic sinus

An autopsy case of spontaneous rupture of a clinically undiagnosed inflammatory aneurysm of the aortic sinus is presented. A 34-year-old man consulted a physician complaining of anterior chest pain and heartburn persisting for 2 days. Diagnoses of common cold and gastritis were made, but 3h later his condition deteriorated, leading to death. Autopsy revealed hemopericardium due to rupture of a saccular aneurysm of the aortic sinus. Histological examination of the aortic sinus disclosed aortitis characterized by fragmentation and loss of elastic tissues with fibrous thickening of the intima. Lymphocytes along with polymorphonuclear leucocytes and plasma cells infiltrated the adventitia and media at severely affected areas. These changes were limited to the aortic sinus, and other portions of the aorta and its main branches were essentially intact. An eccentric intimal thickening with medial degeneration was found within the anterior descending branch of the left coronary artery, and occasional collection of lymphocytes was noted in the epicardium. This case would be a rare and illustrative case for differentiation of aortitis, although we could not pinpoint the etiology of the aortitis because of non-specific histological findings and negative serological results for exogenous pathogens.