Differential Expression Levels of Synaptophysin through Developmental Stages in Cerebral Cortices of Mouse Brain / 체질인류학회지

It is well known that differentiation and growth of central nervous system are accomplished through relatively early stages of development. The formation of neural synapse indicates beginning of electrical signaling between neurons, so that may be a critical step in the differentiation of neurons as well as the development and growth of central nervous system. The purpose of this study was to investigate the differential expression levels and patterns of synaptic marker (synaptophysin) between superficial and deep layers of cerebral cortex according to the developmental stages. We introduced immunofluorescence staining of synaptophysin combined with densitometric analysis for the morphological quantification. The intensities of synaptophysin immuno-reactivities in deep layers of cerebral cortices were significantly higher compared to superficial layers in cerebral cortices of embryonic and neonatal mice. The significant increase of synaptophysin expression in the deep layer of cerebral cortex was mainly confined to the embryonic stage. As the expression of synaptophysin gradually decrease thereafter, the difference of expression level between superficial and deep layers could not find in the adult mice. From this study, we could confirm indirectly through synaptophysin that synaptogenic activities in the deep layer of cerebral cortex shows unique pattern especially during the early stages of brain development. Results from this study will be helpful for understanding different patterns of synaptogenesis among the various regions of developing brain.

Differential Expression Levels of Synaptophysin through Developmental Stages in Cerebral Cortices of Mouse Brain / 체질인류학회지

It is well known that differentiation and growth of central nervous system are accomplished through relatively early stages of development. The formation of neural synapse indicates beginning of electrical signaling between neurons, so that may be a critical step in the differentiation of neurons as well as the development and growth of central nervous system. The purpose of this study was to investigate the differential expression levels and patterns of synaptic marker (synaptophysin) between superficial and deep layers of cerebral cortex according to the developmental stages. We introduced immunofluorescence staining of synaptophysin combined with densitometric analysis for the morphological quantification. The intensities of synaptophysin immuno-reactivities in deep layers of cerebral cortices were significantly higher compared to superficial layers in cerebral cortices of embryonic and neonatal mice. The significant increase of synaptophysin expression in the deep layer of cerebral cortex was mainly confined to the embryonic stage. As the expression of synaptophysin gradually decrease thereafter, the difference of expression level between superficial and deep layers could not find in the adult mice. From this study, we could confirm indirectly through synaptophysin that synaptogenic activities in the deep layer of cerebral cortex shows unique pattern especially during the early stages of brain development. Results from this study will be helpful for understanding different patterns of synaptogenesis among the various regions of developing brain.

Transcriptional targeting of gene expression in breast cancer by the promoters of protein regulator of cytokinesis 1 and ribonuclease reductase 2

For cancer gene therapy, cancer-specific over-expression of a therapeutic gene is required to reduce side effects derived from expression of the gene in normal cells. To develop such an expression vector, we searched for genes over-expressed and/or specifically expressed in cancer cells using bioinformatics and have selected genes coding for protein regulator of cytokinesis 1 (PRC1) and ribonuclease reductase 2 (RRM2) as candidates. Their cancer-specific expressions were confirmed in both breast cancer cell lines and patient tissues. We compared each promoter's cancer-specific activity in the breast normal and cancer cell lines using the luciferase gene as a reporter and confirmed cancer-specific expression of both PRC1 and RRM2 promoters. To test activities of these promoters in viral vectors, the promoters were also cloned into an adeno-associated viral (AAV) vector containing green fluorescence protein (GFP) as the reporter. The GFP expression levels by these promoters were various depending on cell lines tested and, in MDA-MB-231 cells, GFP activities derived from the PRC1 and RRM2 promoters were as strong as that from the cytomegalovirus (CMV) promoter. Our result showed that a vector containing the PRC1 or RRM2 promoter could be used for breast cancer specific overexpression in gene therapy.

A novel way of therapeutic angiogenesis using an adeno-associated virus-mediated angiogenin gene transfer

To develop a novel therapeutic angiogenesis for the treatment of cardiovascular diseases, angiogenin (ANG1) was examined as a potential therapeutic gene. An adeno-associated virus (AAV)-mediated gene delivery system was used to measure the therapeutic efficacy of ANG1. Using a triple co-transfection technique, rAAV-ANG1-GFP, rAAV- VEGF-GFP and rAAV-GFP vectors were produced, which were then used to infect human umbilical vein endothelial cells (HUVECs) in order to evaluate in vitro angiogenic activities. Their protein expressions, tagged with green fluorescent protein (GFP), were monitored by confocal microscopy. The functional activities were measured using wound-healing HUVEC migration assays. The number of migrated cells stimulated by both the expressed ANG1 and the VEGF in rAAV-infected HUVECs increased almost twice the number observed in the expressed GFP control. In vivo angiogenic activities of the expressed ANG1 or VEGF were determined using mouse angiogenesis assays. The angiogenic activities of ANG1 or VEGF expressed in the injected mice were increased by 1.36 and 2.16 times, respectively, compared to those of the expressed GFP control. These results demonstrate that the expressed ANG1 derived from rAAV infection has in vitro and in vivo angiogenic activities and suggest that the rAAV-ANG1 vector is a potential strategy for therapeutic angiogenesis.

Usefulness of High Resolution Computed Tomography (HRCT) in the Diagnosis of Asbestos-Related Lung Diseases / 대한산업의학회지

OBJECTIVES: This study was carried out to improve the medical surveillance program of workers exposed to asbestos by examining the usefulness of High Resolution Computed Tomography (HRCT) in the diagnosis of asbestos-related lung disease. METHODS: The study subjects comprised 162 workers in a ship-repairing yard, 68 of whom had been directly exposed to asbestos and 94 indirectly exposed. The 'Occupational Safety & Health Administration (OSHA) asbestos standard, medical surveillance program' and HRCT were conducted to analyze the aspects of the asbestos-related lung disease. The OSHA asbestos standard, medical surveillance program consists of simple chest x-ray, spirometry and medical questionnaire. RESULTS: Seventeen (10.5%) of the 162 subjects, 10 (14.7%) directly exposed and 7 (7.4%) indirectly exposed, revealed asbestos-related lung disease on HRCT. The sensitivity and specificity of simple chest x-ray for asbestos-related lung disease were 70.6% and 98.6%, and the positive and negative predictive values were 85.7% and 96.6% respectively, as compared with HRCT. HRCT was an effective diagnostic tool especially to detect early asbestos-related lung disease. The study results indicated a relative significant association between the results of spirometry and HRCT. The variables significantly associated with asbestos-related lung disease on HRCT were work duration, smoking history (pack-years), past history of respiratory disease, cough and dyspnea. CONCLUSIONS: In the diagnosis of asbestos-related lung disease, HRCT should be considered not only for workers with positive findings on simple chest x-ray, but also workers with specific findings on spirometry, occupational history, smoking history, and past history of respiratory disease, or with respiratory symptoms such as cough and dyspnea.