• Journal of Microbiology
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• 제42권2호
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• pp.117-125
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• 2004

Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to >1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, rang-ing from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92％ (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-${\alpha}$) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypep-tide reactant was eluted from the gel and was found to cause dose dependent stimulation of the pro-ductions of IL-8 and TNF-${\alpha}$. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

• 대한수의학회지
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• 제29권3호
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• pp.343-359
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• 1989

To study the effects of ibaraki virus on preimplantation mouse embryos collected from prepubertal ICR and BALB/cByJ mice (30~40days old) by superovulation, zona pellucidaintact(ZPI) or free(ZPF) embryos(n=774) of 4- to 8-cell and morulae were exposed to $10^{5.8}$ $TCID_{50}$ of the virus up to 96 hours. The embryos were examined morphologically by observing the degeneration and hatching rates, and virologically and immunologically by determining the presence of infection with the virus, in addition, the effect of washing the embryos to remove virus possibly attached to was also investigated. The ZPI 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rate than those not exposed, for 96, and for 72 to 96 hours, respectively(p<0.01). The ZPF 4- to 8-cell embryos and morulae exposed to the virus showed considerably higher degeneration rates than those not exposed, throughout the whole culture hours in vitro (p<0.01). The ZPI 4- to 8-cell embryos and morulae not exposed to the virus showed considerably higher rates of hatched blastocyst than those exposed (p<0.01). The virus infection rates of the ZPF 4- to 8-cell embryos and morulae were significantly higher than those of the ZPI embryos according to cell culture system. The viral antigen was detected exclusively on the zona pellucida of ZPI embryos, while the antigen was evenly distributed in the blastomeres of ZPF embryos by the immunofluorescent assay. In the ZPI embryos exposed to ibaraki virus, the virus was detected in the two times-washing groups, but not in the ten times-washing groups. The results indicated that zona pellucida of murine embryos would provide an effective protection and that ten times-washing of the ZPI embryos previously exposed to the virus was effective to remove virus from the embryos.

• The Korean Journal of Physiology and Pharmacology
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• 제12권2호
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• pp.51-58
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• 2008

Cardiac fibroblasts constitute one of the largest cell populations in the heart, and contribute to structural, biochemical, mechanical and electrical properties of the myocardium. Nonetheless, their cardiac functions, especially electrophysiological properties, have often been disregarded in studies. $Ca^{2+}$-activated $K^+\;(K_{Ca})$ channels can control $Ca^{2+}$ influx as well as a number of $Ca^{2+}$-dependent physiological processes. We, therefore, attempted to identify and characterize $K_{Ca}$ channels in rat Cardiac fibroblasts. First, we showed that the cells cultured from the rat ventricle were cardiac fibroblasts by immunostaining for discoidin domain receptor 2 (DDR-2), a specific fibroblast marker. Secondly, we detected the expression of various $K_{Ca}$ channels by reverse transcription polymerase chain reaction (RT-PCR), and found all three family members of $K_{Ca}$ channels, including large conductance $K_{Ca}$ (BK-${\alpha}1-\;and\;-{\beta}1{\sim}4$subunits), intermediate conductance $K_{Ca}$ (IK), and small conductance $K_{Ca}$ (SK$1{\sim}4$ subunits) channels. Thirdly, we recorded BK, IK, and SK channels by whole cell mode patch clamp technique using their specific blockers. Finally, we performed cell proliferation assay to evaluate the effects of the channels on cell proliferation, and found that the inhibition of IK channel increased the cell proliferation. These results showed the existence of BK, IK, and SK channels in rat ventricular fibroblasts and involvement of IK channel in cell proliferation.

• 생약학회지
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• 제49권1호
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• pp.47-54
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• 2018

Astilbe rubra (AR) is a perennial, belongs to the Saxifragaceae family, it contains tannin and triterpene. AR has been used in republic korea to improve toxication, fever, pain and convulsion. Recently, number of natural products have been analyzed for potential pharmacological activities including anti-cancer, anti-obesity and anti-diabetic medication. Consequently, we investigated the growth inhibition effect of Astilbe rubra water extract (WAC), ethanol extract (EAC) and bergenin on Hela cell (human adenocarcinoma cell). From whole plant of A. rubra, bergenin was isolated by column chromatography and its structures were identified by $^1H$, $^{13}C$ NMR and IT TOF-ESI MS. High extraction efficiency of bergenin was shown at 0.95% under 60 min reflux extraction with 50% MeOH. The MTS assay showed that EAC (ethanol extract) treatment increased cell death in a dose-dependent manner. Moreover, EAC treatment on Hela cell increased apoptotic cell death and caspase-3 activity. Results suggest that EAC has growth inhibition effect on Hela cells, but not WAC and bergenin. $500{\mu}g/mL$ EAC treatment inhibited Hela cell at $60.2{\pm}1.5%$.

• Journal of Chest Surgery
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• 제54권6호
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• pp.460-465
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• 2021

Background: Metastasis and recurrence of primary cancer are the main causes of cancer mortality. Disseminated tumor cells refer to cancer cells that cause metastasis from primary cancer to other organs. Several recent studies have suggested that circulating tumor cells (CTCs) are associated with the clinical stage, cancer recurrence, cancer metastasis, and prognosis. There are several methods of isolating CTCs from whole blood; in particular, using a membrane filtration system is advantageous due to its cost-effectiveness and availability in clinical settings. In this study, an animal model of lung cancer was established in nude mice using the human large cell lung cancer cell line H460. Methods: Six-week-old nude mice were used. The H460 lung cancer cell line was injected subcutaneously into the nude mice. Blood samples were obtained from the orbital area before cell line injection, 2 weeks after injection, and 2 weeks after tumor excision. Blood samples were filtered using a polycarbonate 12-well Transwell membrane (Corning Inc., Corning, NY, USA). An indirect immunofluorescence assay was performed with the epithelial cell adhesion molecule antibody. The number of stained cells was counted using fluorescence microscopy. Results: The average size of the tumor masses was 35.83 mm. The stained cells were counted before inoculation, 2 weeks after inoculation, and 2 weeks after tumor excision. Cancer cells generally increased after inoculation and decreased after tumor resection. Conclusion: The CTC detection method using the commercial polycarbonate 12-well Transwell (Corning Inc.) membrane is advantageous in terms of cost-effectiveness and convenience.

• Journal of Applied Biological Chemistry
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• 제48권4호
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• pp.198-201
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• 2005

Two known isoquinoline alkaloids, (+)-chelidonine (1) and (-)-stylopine (2), were isolated from $CHCl_3$-soluble fraction of whole plants of Chelidonium majus L. var. asiaticum, and their structures were identified by spectroscopic methods and X-ray crystallographic analysis. Two isolates (1 and 2) were examined for their in vitro cytotoxic activities against five human cancer cell lines including DU-145 (prostate), MCF (breast), A549 (lung), HePG2 (liver), and HT-29 (colon) by sulphorhodamine B (SRB) assay.

• Radiation Oncology Journal
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• 제3권1호
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• pp.1-8
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• 1985

To determine the dose·survival and repair characteristics of the jejunal crypt cells, experimental study was carried out using total 70 mice. Single or split irradiations of 1,100 to 2,200 rad were delivered to whole bodies of $C_{57}$ BL mice, using a cesium 137 animal irradiator and those mice were sacrificed after 90 hours. The number of regenerating crypts per jejunal circumference was counted by a jejunal crypt cell assay technique and dose·response curve was measured. The results were as follows : 1. The average number of jejunal crypts per circumference in control group was 140. In a single irradiation group, the number of regenerated jejunal crypts was, 125, 56, 2 in each subgroup of 1,100 rad, 1,400 rad and 1,800 rad respectively. In split irraiation group, it was 105,44,2 in each subgroup of 1,400rad 1,800rad and 2,200rad respectively. 2. Mean lethal dose of mouse jejunal crypt cell was 167 and 169 rad respectively in a single and split irradiation. 3. Repair dose of sublethal damage was 280 rad. 4. Sublethal damage was completely repaired within 4 hours between the split dose of irradiation.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.96-96
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• 2003

Nek2 (NIMA-related protein) is a mammalian cell cycle-regulated kinase that involves in chromosome condensation and centrosome regulation and NuMA (nuclear mitotic apparatus protein) is involved in spindle assembly during a cell cycle. The cellular distribution and organization of the centrosomal components is completely unknown during fertilization and embryonic development. We examined distribution of two well-known centrosomal proteins, Nek2 and NuMA in mouse gametes and embryos to get an insight in the reorganization of centrosomal proteins during germ cell development and early fertilization. Spermatogenic cells, gametes, and embryos were analyzed with anti-Nek2 or -NuMA antibodies by immunological assay, RT-PCR, and overexpression through gene transfection. Mitotically or meiotically active spermatogenic cells were intensively stained with these antibodies in both centrosomes and cytoplasm, whereas the oocytes showed different staining patterns depending on the meiotic stages. During maturation, GV, GVBD, and MI stage were clearly stained with NuMA antibody in the nucleus or cytoplasm at MII. Also, Nek2 was detectable in cytoplasm as scattered spots or chromosome associated at MII. In early developmental embryo, NuMA was detected in nucleus of each blastomere, while Nek2 was detected in cytoplasm. In contrast to previously reported results, Nek2 and NuMA were detected in both decondensing head, and the centriole of demembranated and decondensed sperm or whole body of trypsin-treated sperm for Nek2. During meiotic progress in oocytes, transcripts levels were the highest in MI stage and then downregulated in MII. Also, it shows dramatically change in early developmental embryos, firstly, it was increased until 4 cell stage and reduced in 8 cell stage, and finally, transcript levels were upregulated until blastoscyst. This finding suggests that cnetrosomal component may play an important role in reorganizing of functional centrosome during fertilization process and subsequent development.

• 약학회지
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• 제40권6호
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• pp.734-738
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• 1996

The development of the alpha2 isoform of (Na,K)ATPase which is high affinity ouabain receptors was studied in the differentiating nonfusing muscle cell line BC3H-1. T he differentiation process of BC3H-1 cell line was confirmed by 2-dexy-D-[$^3$H] glucose uptake experiment and the quantity of the expression of ${\alpha}_2$ isoform was measured using a whole cell [$^3$H] ouabain-binding assay. Undifferentiated growing BC3H-1 cells, myoblasts, exhibited low levels of insulin-stimulated glucose uptake and [$^3$H] ouabain-binding sites. In contrast, differentiated BC3H-1 cells, myocytes, had a 5.6-fold increase in insulin-stimulated glucose uptake and 5-fold increase in [$^3$H] ouabain-binding sites. Scatchard analysis showed that myocytes developed more [$^3$H] ouabain-binding sites than myoblasts vath a dissociation constant (kd) of 6${\times}10^{-8}$M and capacity of 6.l${\times}10^{-5}$ sites/cell. Therefore. it seems that myoblasts express low levels of ${\alpha}_2$ subunit and probably the majority of ${\alpha}_1$ subunit, whereas myocytes express high levels of ${\alpha}_2$ isoform. The results indicate that the expression of ${\alpha}_2$ isoform is developmentally regulated during differentiation and that BC3H-1 culture system provides an excellent model for the study of differentiation and mechanism of (Na,K)ATPase action in muscle which requires electrical excitability.

• Molecules and Cells
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• 제39권11호
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• pp.807-813
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• 2016

Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of $1.3{\times}10^6CFU/ml$. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate ($5{\times}10^4CFU/ml$). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of $2{\times}10^4CFU/mL$ of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.