• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.888-897
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• 2009

Lipase diversity in glacier soil was assessed by culture-independent metagenomic DNA fragment screening and confirmed by cell culture experiments. A set of degenerate PCR primers specific for lipases of the hormone-sensitive lipase family was designed based on conserved motifs and used to directly PCR amplify metagenomic DNA from glacier soil. These products were used to construct a lipase fragment clone library. Among the 300 clones sequenced for the analysis, 201 clones encoding partiallipases shared 51-82% identity to known lipases in GenBank. Based on a phylogenetic analysis, five divergent clusters were established, one of which may represent a previously unidentified lipase subfamily. In the culture study, 11 lipase-producing bacteria were selectively isolated and characterized by 16S rDNA sequences. Using the above-mentioned degenerate primers, seven lipase gene fragments were cloned, but not all of them could be accounted for by the clones in the library. Two full-length lipase genes obtained by TAIL-PCR were expressed in Pichia pastoris and characterized. Both were authentic lipases with optimum temperatures of ${\le}40^{\circ}C$. Our study indicates the abundant lipase diversity in glacier soil as well as the feasibility of sequence-based screening in discovering new lipase genes from complex environmental samples.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.9
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• pp.4433-4437
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• 2016

Background: Gastric cancer (GC) is the fifth most common cancer worldwide. Since development is usually asymptomatic, it is generally diagnosed at an advanced stage. The value of screening in patients with nonspecific symptoms for GC is controversial. Aim: The study aimed to evaluate whether hematological parameters (platelet count (PC), mean platelet volume (MPV), MPV/PC ratio, red blood cell distribution width (RDW), neutrophil to lymphocyte ratio (NLR), and platelet to lymphocyte ratio (PLR)) are useful markers to differentiate between gastric cancer patients and healthy individuals. Materials and Methods: Sixty-one patients with gastric cancer and sixty-one healthy individuals were enrolled to the survey and retrospective analysis of selected blood parameters were performed. Results: The mean values of PC, MPV, RDW, NLR, and PLR were significantly higher in GC patients compared to the control group. No statistical differences were observed in MPV/PC ratios. Likewise, no significant statistical differences were revealed in values of blood parameters among TNM stage groups. The RDW showed the highest diagnostic specificity and sensitivity. Conclusions: Hematological parameters: PC, MPV, RDW, NLR, PLR have diagnostic power and can discriminate patients with gastric cancer from patients without cancer. Blood parameters compared with clinical symptoms might alert physicians and patients and lead to performancce of upper gastrointestinal endoscopy, the gold standard in gastric cancer screening and therebly increase the early detection of cancer.

• Journal of mucopolysaccharidosis and rare diseases
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• v.4 no.1
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• pp.14-20
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• 2018

Mucopolysaccharidoses (MPSs) are a group of rare inherited metabolic disorders caused by specific lysosomal enzyme deficiencies leading to the sequential degradation of glycosaminoglycans, causing substrate accumulation in various cells and tissues and progressive multiple organ dysfunction. The rare disease medical care team at Mackay Memorial Hospital in Taiwan has been dedicated to the study of MPSs for more than 20 years. Since 1999, more than 50 academic papers focusing on MPSs have been published in international medical journals. Topics of research include the following items regarding MPSs: incidence, natural history, clinical manifestations, gene mutation characteristics, cardiac function, bone mineral density, sleep studies, pulmonary function tests, hearing assessments, percutaneous endoscopic gastrostomy, anesthetic experience, imaging analysis, special biochemical tests, laboratory diagnostics, global expert consensus conferences, prenatal diagnosis, new drug clinical trials, newborn screening, and treatment outcomes. Of these published academic research papers, more than half were cross-domain, cross-industry, and international studies with results in cooperation with experts from European, American and other Asian countries. A cross-specialty collaboration platform was established based on high-risk population screening criteria with the acronym "BECARE" (Bone and joints, Eyes, Cardiac and central nervous system, Abdomen and appearance, Respiratory system, and Ear, nose, and throat involvement). Through this platform, orthopedic surgeons, rheumatologists, ophthalmologists, cardiologists, rehabilitation physicians, gastroenterologists, otorhinolaryngologists, and medical geneticists have been educated with regards to awareness of suspected cases of MPSs patients to allow for a further confirmative diagnosis of MPSs. Because of the progressive nature of the disease, an early diagnosis and early multidisciplinary therapeutic interventions including surgery, rehabilitation programs, symptom-based treatments, hematopoietic stem cell transplantation, and enzyme replacement therapy, are very important.

• Animal cells and systems
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• v.6 no.4
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• pp.317-322
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• 2002

To improve conventional yeast one-hybrid screening, we have developed an efficient mammalian one-hybrid system that allows rapid isolation of com-plementary DNAs which are able to induce human p14$^{ARF}$. tumor suppressor gene. A 1.5 kb promoter region of p14$^{ARF}$ was fused to EGFP to generate ARF promoter-EGFP reporter vector. This reporter plasmid was stably trans-fected into NIH3T3 cells for generation of reporter cell line. When the reporter cell line was infected with E2F-1 together with excess amounts of empty vector, the cells that received the positive modulator were readily identifiable by green fluorescence using FACS. The GFP-positive cells were cloned directly from the cultured cells and expanded in bulk culture. The genomic DNAs from GFP-positive cells were prepared and the CDNA insert in integrated retroviral genome was recovered by PCR using primers annealing to the retroviral vector sequences flanking the insert-cloning site. This system should be useful for efficient screening of expression CDNA libraries in mammalian cells to identify novel upstream regulators for spe-cific genes by one-hybrid interaction.ion.

• The Korean Journal of Physiology and Pharmacology
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• v.10 no.5
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• pp.263-271
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• 2006

Since astrocytes were shown to play a central role in maintaining neuronal viability both under normal conditions and during stress such as ischemia, studies of the astrocytic response to stress are essential to understand many types of brain pathology. The micro array system permitted screening of large numbers of genes in biological or pathological processes. Therefore, the gene expression patterns in the in vitro model of astrocytes following exposure to oxygen-glucose deprivation (OGD) were evaluated by using the micro array analysis. Primary astrocytic cultures were prepared from postnatal Swiss Webster mice. The cells were exposed to OGD for 4 hrs at $37^{\circ}C$ prior to cell harvesting. From the cultured cells, we isolated mRNA, synthesized cDNA, converted to biotinylated cRNA and then reacted with GeneChips. The data were normalized and analyzed using dChip and GenMAPP tools. After 4 hrs exposure to OGD, 4 genes were increased more than 2 folds and 51 genes were decreased more than 2 folds compared with the control condition. The data suggest that the OGD has general suppressive effect on the gene expression with the exception of some genes which are related with ischemic cell death directly or indirectly. These genes are mainly involved in apoptotic and protein translation pathways and gap junction component. These results suggest that microarray analysis of gene expression may be useful for screening novel molecular mediators of astrocyte response to ischemic injury and making profound understanding of the cellular mechanisms as a whole. Such a screening technique should provide insights into the molecular basis of brain disorders and help to identify potential targets for therapy.

• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.2
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• pp.86-92
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• 2004

In this paper miniaturized disposable micro/nanofluidic components applicable to bio chip, chemical analyzer and biomedical monitoring system, such as blood analysis, micro dosing system and cell experiment, etc are reported. This system includes various microfluidic components including a micropump, micromixer, DNA purification chip and single-cell assay chip. For low voltage and low power operation, a surface tension-driven micropump is presented, as well as a micromixer, which was implemented using MEMS technology, for efficient liquid mixing is also introduced. As bio-reactors, DNA purification and single-cell assay devices, for the extraction of pure DNA from liquid mixture or blood and for cellular engineering or high-throughput screening, respectively, are presented.

• Journal of Korean Dental Science
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• v.4 no.1
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• pp.33-37
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• 2011

Gorlin-Goltz syndrome is an autosomal dominant disorder with a high degree of penetrance. It is characterized by basal cell carcinomas, odontogenic keratocysts, palmar and plantar pits and ectopic calcifications of falx cerebri. The presence of two major and one minor criteria or one major and three minor criteria are necessary to establish a diagnosis. Early diagnosis and treatment of Gorlin-Goltz syndrome, as well as family screening and genetic counseling are essential as it may be associated in 10% of patients with aggressive basal cell carcinoma and malignant neoplasias. We report here a patient with Gorlin-Goltz syndrome.

• Biomedical Science Letters
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• v.12 no.4
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• pp.451-455
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• 2006

It is demonstrated that inorganic mercury has cytotoxic effect on glial cells. Recently, oxygen radicals is involved in methylmercuric chloride (MMC)-induced cytotoxicity. But, the toxic mechanism of MMC is left unknown. The purpose of this study was to examine the cytotoxicity of MMC on $C_6$ glioma cells. The cytotoxicy was measured by cell viability using XTT assay in $C_6$ glioma cells. Colorimetric assay is regarded as a very sensitive screening method for the determination of the cell viability on various agents. In this study, MMC decreased cell viability according to the dose- and time dependent manners after $C_6$ glioma cells were grown with various concentrations of MMC for 48 hours. In the protective effect of allopurinol on MMC-induced cytotoxicity, allopurinol was effective in the prevention of MMC-induced cytotoxicity in these cultures. These results suggest that MMC has highly cytotoxic effect on $C_6$ glioma cells by the decrease of cell viavility, and free radical scavenger such as allopurinol was effective on organic mercury-induced cytotoxicity in these cultures.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.sup1
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• pp.126-129
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• 2008

Renal cell carcinoma is the most common histological type of renal malignancy, predominant in men and the primary treatment modality of this tumor is surgery. The role of diagnostic imaging in the management of this tumor is the evaluation of extent of disease as well as the detection and characterization of renal mass. US has long been a routine screening tool for kidney but tomographic imaging modalities such as CT and MRI begin to be also commonly used these days. On the other hand, the sensitivity of $^{18}F-FDG-PET$ in detection of renal mass is relatively low because of inherent limitation caused by FDG excretion pathway despite avid uptake of FDG to tumor cell per se. Many studies revealed FDG PET scan could play an important role in detection of metastatic lesions although the sensitivity for the detection of primary lesion is not so high. Furthermore, development of PET/CT scanner will make it possible to expand the indication of FDG PET scan in this malignancy.

• Korean Journal of Animal Reproduction
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• v.17 no.1
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• pp.13-19
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• 1993

We investigated the effects of CP-2 extracted from the mixture of Copis and Croton tiglium L, which showed very high cytotoxic effect to the various tumor cells, on the growth and steroidenesis of primary and transformed cell lines PA-GS6 and PO-GRS1 by cotransfectionwith SV40 and Ha ras oncogenes. CP-2 inhibited the growth of PA-GS6 and PO-GRS1 in a dose dependent manner when the growth of them was measured by cell number and by protein content, while CP-2 did not affect the growth of primary granulose cells. Productions of progesterone ofprimary and transformed granulosa cells were stimulated by forskolin, but this stimulatory effect was blocked by treatment of CP-2. Clinical application of CP-2 asa new anti-cancer drug and utilization of transformed granulosa cells as a model system for the screening of anti-cancer drug were also discussed.