• Journal of Nutrition and Health
• /
• v.41 no.3
• /
• pp.224-231
• /
• 2008

We examined the effect of indole-3-carbinol (I3C, $C_9H_9NO$), an autolysis product of a glucosinolate and a glucobrassicin in vegetables, on MMP-2, -9 activities and TIMP-l and -2 inductions via microtubule-associated protein kinase (MAPK) signaling pathway in prostate cancer cell line, PC3 cells. Our results indicated that I3C inhibited cell growth of PC3 cells in dose (0,50, 100 ,${\mu}M$) and time (0,24,48 and 72 h) dependent manners. Using gelatin zymography for MMP activity, we demonstrated that I3C significantly decrease MMP-2 and -9 activities in PC3 cells. We also observed that I3C decreased the proteins and mRNA levels of MMP-2 and -9 in PC3 cells as well. Inversely, expressions of TIMP-l and -2 protein and mRNA in PC3 cells were increased by I3C in a dose dependent manner. In another experiment, we showed that I3C inhibited PC3 cells invasiveness by using marigel invasion assay and we also found that I3C suppressed MMP transcriptional activity by MAPK signaling pathways. Taken together, our results suggest that I3C may contribute to the potential beneficial food component to prevent the cancer metastasis in prostate cancer cells. (KoreanJNutr2008; 41(3): 224~23I)

• Proceedings of the KOSOMBE Conference
• /
• v.1997 no.11
• /
• pp.357-360
• /
• 1997

For medical students and doctors, knowledge of the 3-dimensional (3D) structure of the heart is very important in diagnosis and treatment of the heart diseases. 2-dimensional (2D) tools (e.g. anatomy book) or classical 3D tools (e.g. plastic model) are not sufficient or understanding the complex structures of the heart. Moreover, it is not always guaranteed to dissect the heart of cadaver when it is necessary. To overcome this problem, virtual dissection systems of the heart have been developed. But these systems are not satisfactory since they are made of radiographs; they are not true 3D images; they can not be used to dissect freely; or they can only be operated on the workstation. It is also necessary to make the dissection systems incorporating the various races and tribes because of the organ's difference according to race and tribe. This study was intended to make the 3D image of the heart from a Korean cadaver, and to establish a virtual dissection system of the heart with a personal computer. The procedures or manufacturing this system were as follows. 1. The heart from a Korean adult cadaver was embedded with gelatin solution, and serially cross-sectioned at 1mm-thickness on a meat slicer. Pictures or 153 cross-sectioned specimens were inputted into the computer using a digital camera ($756{\times}504$ resolution, true color). 2. The alignment system was established by means of the language of IDL, and applied to align 2D images of the heart. In each of 2D images, closed curves lining clean and dirty blood pathways were drawn manually on the CorelDRAW program. 3. Using the language of IDL, the 3D image and the virtual dissection system of the heart were constructed. The virtual dissection system of the heart allowed or ree rotation, any-directional sectioning, and selected visualization of the heart's structure. This system is expected to become more advanced, and to be used widely through Internet or CD-title as an educational tool for medical students and doctors.

• Proceedings of the KOSOMBE Conference
• /
• v.1998 no.11
• /
• pp.57-59
• /
• 1998

For medical students and doctors, knowledge of the three-dimensional (3D) structure of brain is very important in diagnosis and treatment of brain diseases. Two-dimensional (2D) tools (ex: anatomy book) or traditional 3D tools (ex: plastic model) are not sufficient to understand the complex structures of the brain. However, it is not always guaranteed to dissect the brain of cadaver when it is necessary. To overcome this problem, the virtual dissection programs of the brain have been developed. However, most programs include only 2D images that do not permit free dissection and free rotation. Many programs are made of radiographs that are not as realistic as sectioned cadaver because radiographs do not reveal true color and have limited resolution. It is also necessary to make the virtual dissection programs of each race and ethnic group. We attempted to make a virtual dissection program using a 3D image of the brain from a Korean cadaver. The purpose of this study is to present an educational tool for those interested in the anatomy of the brain. The procedures to make this program were as follows. A brain extracted from a 58-years old male Korean cadaver was embedded with gelatin solution, and serially sectioned into 1.4 mm-thickness using a meat slicer. 130 sectioned specimens were inputted to the computer using a scanner ($420\times456$ resolution, true color), and the 2D images were aligned on the alignment program composed using IDL language. Outlines of the brain components (cerebrum, cerebellum, brain stem, lentiform nucleus, caudate nucleus, thalamus, optic nerve, fornix, cerebral artery, and ventricle) were manually drawn from the 2D images on the CorelDRAW program. Multimedia data, including text and voice comments, were inputted to help the user to learn about the brain components. 3D images of the brain were reconstructed through the volume-based rendering of the 2D images. Using the 3D image of the brain as the main feature, virtual dissection program was composed using IDL language. Various dissection functions, such as dissecting 3D image of the brain at free angle to show its plane, presenting multimedia data of brain components, and rotating 3D image of the whole brain or selected brain components at free angle were established. This virtual dissection program is expected to become more advanced, and to be used widely through Internet or CD-title as an educational tool for medical students and doctors.

• Korean Journal of Microbiology
• /
• v.35 no.2
• /
• pp.158-163
• /
• 1999

A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

• Korean Journal of Food Science and Technology
• /
• v.39 no.6
• /
• pp.619-624
• /
• 2007

Sedum sarmentosum, also blown as stonecrop (dolnamul), is a widely consumed herb, and is used as an ingredient in salads in Korea. Unfortunately, sedum is perishable and vulnerable to tissue damage during preservation. Therefore, this feasibility study was performed in order to increase the availability of sedum and increase its value. Various concentrations of sedum extract (0.5-3%) were added to gelatin jelly, and their physicochemical properties were determined. The ascorbic acid content of the sedum jelly increased in proportion to the sedum extract concentration. Calcium content of the sedum jelly was 4 to 28 times higher than that of the control. Contrary to the control, iron was detected in the sedum jelly (0.023-1.031 mg/100 g dry weight). Furthermore, magnesium and potassium levels were higher in the sedum extract groups. There were significant differences (p < 0.05) in greenness (a value) and yellowness (b value) between the control and the sedum extract groups. However, significant differences between the 2% and 3% sedum extract groups were not detected. As sedum extract concentration increased, the pH level of soft jellies (solid state) decreased. Therefore, hardness and gumminess were decreased significantly. These results are in agreement with the sensory evaluation. According to sensory tests, the values for palatability, appearance, and color in the 2% sedum extract group were higher than those of the 0.5-1% and 3% sedum extract groups.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.47-52
• /
• 2006

This study was conducted to examine whether the parthenogenetic mouse embryonic stem (P-mES) cells can differentiate into functional cardiomyocytes in vitro similar to (mES) cells. p-mES04 and IVF-derived mES03 cells were cultured by suspension culture for 4 days. The formed embryoid bodies (EBs) were treated with 0.75% dimethyl-sulfoxide (DMSO) for further 4 days (4-/4+), and then plated onto gelatin coated culture dish. The appearance of contracting cardiomyocytes from the P-mES04 and mES03 cells was examined for 30 days. The highest cumulative frequency was detected at days 13 (69.83%) and 22 (61.3%), respectively. By immunocytochemistry, beating P-mES04 cells were positively stained with muscle specific anti-sarcomeric a-actinin Ab and cardiac specific anti-cardiac troponin I Ab similar to contracted mES03 cells. When the expression of cardiac muscle-specific genes was analyzed by RT-PCR, beating P-mES04 cells were expressed cardiac specific L-type calcium channel, a1C, cardiac myosin heavy chain a, cardiac muscle heavy polypeptide $7{\beta}$, GATA binding protein 4 and atrial natriuretic factor, but not expressed skeletal muscle specific L-type calcium channel, a1S, which was similar to male adult heart cells and mES03-derived beating cardiomyocytes. The result demonstrates that the P-mES cells can be used as an alternative for the study on the characteristic analysis of in vitro cardiomyocyte differentiation from the ES cells.

• Clinical and Experimental Reproductive Medicine
• /
• v.25 no.2
• /
• pp.161-170
• /
• 1998

Protein expression patterns of matrix metalloproteinases (MMPs) were examined in mouse reproductive organs during estrous cycle. Estrous cycle was classified into diestrus, proestrus, estrus or metestus and MMP expression was analyzed by zymography using gelatin as a substrate. Uterine fluid (UF) obtained both at diestrus and proestrus exhibited 4 major MMPs including 106kDa, 64kDa, 62kDa and 59kDa gelatinases. However, in UF at estrus, the gelatinolytic activity of 64kDa MMP disappeared and that of 106kDa and 62kDa MMPs dramatically decreased. At metestrus, 64kDa MMP activity reappeared and 106kDa and 62kDa MMP exhibited increased activities such that the band intensity of 106kDa was comparable to that in UF at diestrus. Gelatinolytic activity of 59kDa MMP was not changed throughout the cycle. Both ovarian and oviductal tissue homogenate revealed 4 MMPs which corresponded to the 4 MMPs of UF. However, unlike UF MMPs, gelatinolytic activity of these MMPs did not show distinct changes throughout the cycle. Either an inhibitor of MMP, 1,10-phenanthroline, or a metal chelator, EDTA, abolished the appearance of the above MMP activities in gelatinated gel whereas a serine proteinase inhibitor, phcnylmethylsulfonyl fluoride, failed to inhibit the appearance of MMP activities, proving that gelatinolytic activity of the above reproductive tissues were due to the enzymatic activity of MMP. When gclatinolytic activity of mouse serum was examined, it revealed 5 MMPs (131kDa, 106kDa, 89kDa, 64kDa and 62kDa bands) and one gelatinase (84kDa) band. From these results, it is concluded that the protein expression of MMPs of mouse reproductive organs, particularly uterus, is temporally regulated during estrous cycle and uterine 106kDa, 64kDa and 62kDa MMPs are suggested to play an important role in cyclic tissue remodeling of mouse uterus.

• Food Science of Animal Resources
• /
• v.22 no.3
• /
• pp.259-266
• /
• 2002

The egg shell membrane degrading isolated from soil was identified as Bacillus licheniformis by 16S rDNA identification method. A keratinase was isolated from the Baciilu licheniformis culture. DEAE-cellulose ion-exchange and Sephadex C-75 gel chromatograhies were used to purify the enzyme. The specific activity was increased 17.3-fold by the purification procedures. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and Sephadex G-75 chromatography indicated that the purified keratinase was monomeric and had a molecular weight of 65 kDa. The enzyme showed optimum activity at pH 9.0, and was stable above pH 9.0. The optimum temperature was 50$\^{C}$ and the enzyme was stable in the temperature ranges from 20$\^{C}$ to 50t. By the addition of 1 mM and 10 mM FeSO4, the activities of the enzyme were increased to 111$\pm$4.6% and 133$\pm$3.79%, respectively. The keratinase was an alkaline serine pretense because it was inhibited only by phenylmethylsulfonylfluorice (PMSF).

• Journal of Microbiology and Biotechnology
• /
• v.16 no.9
• /
• pp.1347-1354
• /
• 2006

The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.

• Tuberculosis and Respiratory Diseases
• /
• v.77 no.2
• /
• pp.73-80
• /
• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.