Observation of 17 Asphyxial Suicides by Helium Gas / 대한법의학회지

Since the book "Final Exit: The Practicalities of Self-Deliverance and Assisted Suicide for the Dying" was published in 1991, there has been a worldwide increase in the number of cases of suicidal asphyxiation using helium gas. However, no domestic reports have been published thus far. Recently, we encountered a case of asphyxial suicide by inhalation of helium from inside a plastic bag. Subsequently, we reviewed the records of the Scientific Crime Analysis System of National Police Agency of Korea. There were 17 cases of helium-associated asphyxial suicides between January 1, 2000 and June 30, 2012. The average age of the deceased was 30.6 years with a male to female ratio of approximately 5:1. Thirteen of 17 such deaths occurred in the victims'houses. In all cases, the method of death involved placing a plastic bag over the head with a hose attached to helium tanks. In 6 of 17 cases, the decedents had psychiatric disorders such as depression. As neither characteristic signs of death nor conventional methods for detecting helium gas exist, a thorough investigation of the death scene and the decedents'environment is extremely vital for confirming death due to helium asphyxiation.

An Immuno-Electron Microscopic Study for Apoptosis and Expression of p53 on the Synovium of the Osteoarthritic Knee in Human / 대한해부학회지

This study was designed to observe the apoptosis and expression of p53 in the osteoarthritic synovial membrane compared with normal synovial membrane of human. The collected normal and osteoarthritic synovia were dissected and fixed for two hours (in 4% paraformaldehyde and 0.1% glutaraldehyde solution). In this study, TUNEL staining and immunocytochemical gold labeling techniques were used. In the immunocytochemical gold labeling techniques, primary antibodies which was to be monoclonal mouse anti-p53 were used. Donkey anti-mouse IgG tagged with 6 nm colloidal gold particles was used as the secondary antibody. The tissues were observed under JEOL 1200 EX-II transmission electron microscope. The results were as follows. 1. On TUNEL staining, normal synovium were not seen TUNEL positive signal cells. But, in the osteoarthritic synovium, few TUNEL positive cells were seen in synovial membrane and subsynovial layers. 2. On the transmission electron microscopic observation, normal synovium had 1~3 synovial cell layers, which had phagocytic synovial cells and secretory synovial cells. The osteoarthritic synovium had 2~5 synovial cell layers, which consisted with abnormally proliferated secretory synovial cells. These cells had heterochromatin in nucleus and well developed endoplasmic reticulum in the cytoplasm. 3. On the normal synovium of the human knee joint, p53 positive cells were not identified. But, in the osteoarthritic synovium of the human knee joint, p53 positive cells were identified. These cells were recognized secretory synovial cells and apoptotic cells. In the secretory synovial cells, the distributions of p53 were mitochondria and rough endoplasmic reticulum. In the apoptotic cells, p53 were marked on rough endoplasmic reticulum, which showed secretory synovial cells. On the basis of above findings, it is obvious that osteoarthritic synovial membrane has identified the apoptotic cells compared with normal synovium. These apoptotic cells might be identified as mainly secretory synovial cells and a few phagocytic synovial cells. The immunogold of p53 was marked at rough endoplasmic reticulum and in nucleus of apoptotic cells. Apoptosis in the osteoarthritic synovium seemed to be developed through p53 negative dependent pathway.

An Immunoelectronmicroscopic Study on the Cellular Expression of Extracellular Matrix Components during Skin Regeneration after Damage in the Mouse / 대한해부학회지

To observe the cellular expression of extracellular matrix components during mouse skin regeneration, the wounded skin samples were processed by immunoelectronmicroscopic methods, using primary antibodies for fibronectin, collagen type IV and laminin. The tissues were observed under transmission electron-microscope. The results were summarized as follows. 1. The granulation tissues and x-cells were observed in the wound margin at 18 hr post injury. The number of fibroblasts was increased in the granulation tissues at 1 day post injury. 2. The expression of fibronectin was observed in x-cells at 18 hr post injury, and in fibroblasts at 1 day post injury. In x-cells, after 1 day post injury, the expression of fibronectin was decreased. 3. At 1 day post injury, the expression of collagen type IV was increased in fibroblasts whereas not in x-cells. 4. The expression of laminin was increased by 18 hr post injury, but decreased after 1 day post injury. On the basis of above findings, in mouse, the regenerations of wounded skin were faster than other animals. The first step, infiltration response, was processed till 18 hr or 1 day post injury. The second step, fibroblast proliferation phase, began at 1 day post injury. In the regenerations of wounded skin, x-cells migrated to the wound region and activated, earlier than fibroblast. Thereafter, x-cells which appeared to be transformed into fibroblasts, played an important role in the synthesis of fibronectin and collagen type IV, and the formation of granulation tissue, with migrated fibroblasts to the wound region.

An Immuno -Electron Microscopic Study for Perlecan in the Synovium of Degenerative Arthritis of Knee / 대한해부학회지

This study was designed to observe the expression of perlecan in the normal and degenerative arthritic synovial membrane. By using the immunohistochemical staining and immuno -electron microscopical gold labeling techniques, we observed five materials of normal and degenerative arthritic synovia each. The results were as follows. 1. By the immunohistochemical methods, perlecan -positive staining was seen on the 1 ~2 cell layers of the normal synovial membrane. But, a weaker staining compared to that seen in the normal synovial membrane was found in the degenerative arthritic synovial membrane. 2. Under the electron microscopic observation, perlecan was largely distributed in the rough endoplasmic reticulum of the secretory synovial cell, and in the vacuoles of the phagocytic synovial cell on the normal synovium of the human knee joint. It was also found in the extracellular matrix of the synovial membrane. 3. Perlecan -positive cells were also identified on the degenerative arthritic synovium of the human knee joint. However, fewer perlecan was observed here than that found in the normal synovium. In conclusion, perlecan is synthesized by the secretory synovial cells and degraded by the phagocytic synovial cells. And it, known as a major component of the basement membrane, also proven to exist in the extracellular matrix of the synovial membrane having no basement membrane. From the fact that less perlecan was observed in the degenerative arthritis, perlecan is might to play a major role in the degenerative process.

Histochemical Study on the Expression of Heat Shock Protein 70 mRNA in Developing Mouse Brain / 대한해부학회지

Using in situ hybridization technique with digoxigenin-labelled riboprobe, study on the expression of hsp 70 mRNA in the developing mouse brain was performed. The results obtained are as follows; 1. In embryonic day 16 group, cells with strong reactivity to hsp70 mRNA were found in spinal cord. In neuroepithelial layer lining fourth ventricle and external granular layer of cerebellum, moderate reactivity was observed. But the reactivity was weak in the forebrain including cerebral cortex, diencephalon and olfactory bulb. 2. In embryonic day 18 group, the regional pattern of hsp70 mRNA expression was similar to that of embryonic day 16 group. In medulla oblongata, however, stronger reactivity was found in the embryonic day 18 group. 3. In postnatal day 0 mice group, cells with moderate or strong reactivity to hsp70 mRNA were found in the overall area of central nervous system, Especially, cells with moderate reactivity were found in the dentate gyrus of hippocampus, and the supragranular cortical plate and subplate neocortex. 4. In postnatal day 2 mice group, cells with moderate or strong reactivity to hsp70 mRNA were found in the same pattern as in postnatal day 0 group. Further differentiation of cerebral cortex and cerebellum was found. 5. Strong expression of hsp70 mRNA was found in the areas with high rate of cell division. In general, the area of expression moved to more rostral area in central nervous system as development proceeds. Above results suggest that hsp70 play an important role in the development and differentiation of central nervous system.

The Electron Microscopic Study on the Development of Knee Joint in Rat / 대한해부학회지

These study was designed to observe the appearance and the characteristics of apoptotic cells during the development of knee joint in rat. The fetus were collected on the 16th, 17th, 18th, 19th, and 20th day of pregnancy. In this study, TUNEL staining, electron microscopic investigation and immunocytochemical gold labeling techniques were used. In the immuno-cytochemical gold labeling techniques, primary antibodies were used, which were to be polyclonal rabbit anti-mouse/ rat Bax, polyclonal rabbit anti-tissue transglutaminase C, and polyclonal goat anti-cpp32p20. The samples were observed under JEOL 1200 EX-II transmission electron microscope. The results were as follows. 1. In a 16-day-old fetus, between femur and tibia cartilages, mesenchymal cells were observed. Mesenchymal cells had marginated heterochromatin and dilated rough endoplasmic reticulum. 2. In a 17-day-old fetus, the knee joint clefts were first formed. In the primordial cruciate ligaments between the cartilages, capillaries were scattered. The apoptotic cells, which had fragmented and condensed nucleus, showed in the synovium. And necrotic cells, which had nuclear chromatin margination, perinuclear cisternae, and dilated rough endoplasmic reticulum, also were observed in the joint cleft surface. 3. From the 18-day-old fetus, phagocytic synovial cells and secretory synovial cells could be confirmed. The apoptotic cells were not seen. 4. In a 17-day-old fetus, a few cells were positive for TUNEL reaction in the joint cleft region. 5. In a 17-day-old fetus, Bax were marked on the mitochondria, endoplasmic reticulum of apoptotic cells. Also, it was marked at the phagocytosed apoptotic bodies in the neighboring cells. 6. In a 17-day-old fetus, the tissue Transglutaminase C were marked in the perinuclear region, vacuoles, cell membrane and extracellular matrix of the apoptotic cells. Also, it was marked at the phagocytosed apoptotic bodies in the neighboring cells. 7. In a 17-days-old fetus, CPP32 labeling were marked in the cytoplasm of the apoptotic cells. Practically, it was distributed between the phagocytosed apoptotic bodies and the neighboring cells. On the basis of above findings, it is obvious that the joint cleft are first formed in a 17-day-old fetus, a few cells are to be TUNEL positive signals, and the apoptotic cells contain Bax, tissue Transglutaminase C, and CPP32. Therefore the apoptotic cells and the necrotic cells are appeared in the 17-day-old fetus, and these cells are concerned with joint cleft formation.