• Biomolecules & Therapeutics
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• v.22 no.6
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• pp.540-546
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• 2014

The high mortality rates associated with cancer reflect the metastatic spread of tumor cells from the site of their origin. Metastasis, in fact, is the cause of 90% of cancer deaths. Therefore, considerable effort is being made to inhibit metastasis. In the present study, we screened ketotifen for anti-migratory and anti-invasive activities against MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer cells. Cancer cell migration and invasion were measured using multi-well chambers. Additionally, western blots were used to examine the effects of ketotifen on the expressions of CDC42, Rho, Rac, and matrix metalloproteinase 9 (MMP-9). The results showed that ketotifen dose-dependently suppressed the migration and invasion of MDA-MB-231 and HT-1080 cells. Ketotifen also suppressed the expressions of CDC42, Rac, and Rho, which, significantly, are involved in MDA-MB-231 and HT-1080 cancer cell migration. Moreover, ketotifen suppressed the expression and activity of MMP-9, which is involved in degradation of the extracellular matrix leading to invasion. The overall data suggested that ketotifen suppresses the migration and invasion of MDA-MB-231 and HT-1080 cancer cells via inhibition of CDC42, Rac, Rho, and MMP-9 expression.

• Tuberculosis and Respiratory Diseases
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• v.82 no.2
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• pp.151-157
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• 2019

Background: Tuberculosis (TB) is the most important disease screened for upon patient history review during preimmigration medical examinations as performed in South Korea in prospective immigrants to certain Western countries. In 2007, the U.S. Centers for Disease Control and Prevention (CDC) changed the TB screening protocol from a smear-based test to the complete Culture and Directly Observed Therapy Tuberculosis Technical Instructions (CDOT TB TI) for reducing the incidence of TB in foreign-born immigrants. Methods: This study evaluated the effect of the revised (as compared with the old) protocol in South Korea. Results: Of the 40,558 visa applicants, 365 exhibited chest radiographic results suggestive of active or inactive TB, and 351 underwent sputum tests (acid-fast bacilli smear and Mycobacterium tuberculosis culture). To this end, using the CDOT TB TI, 36 subjects (88.8 per $10^5$ of the population) were found to have TB, compared with only seven using the older U.S. CDC technical instruction (TI) (p<0.001). In addition, there were six drug-resistant cases which were identified (16.7 per $10^5$ of the population), two of whom had multidrug-resistance (5.6 per $10^5$ of the population). Conclusion: The culture-based 2007 TI identified a great deal of TB cases current to the individuals tested, as compared to older U.S. CDC TI.

• Journal of Microbiology and Biotechnology
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• v.34 no.7
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• pp.1425-1432
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• 2024

This study analyzed the effects of Ca²⁺ metal ions among culture medium components on the Chlorella sorokiniana strain DSCG150 strain cell growth. The C. sorokiniana strain DSCG150 grew based on a multiple fission cell cycle and growth became stagnant in the absence of metal ions in the medium, particularly Ca²⁺. Flow cytometry and confocal microscopic image analysis results showed that in the absence of Ca²⁺, cell growth became stagnant as the cells accumulated into four autospores and could not transform into daughter cells. Genetic analysis showed that the absence of Ca²⁺ caused upregulation of calmodulin (calA) and cell division control protein 2 (CDC2_1) genes, and downregulation of origin of replication complex subunit 6 (ORC6) and dual specificity protein phosphatase CDC14A (CDC14A) genes. Analysis of gene expression patterns by qRT-PCR showed that the absence of Ca²⁺ did not affect cell cycle progression up to 4n autospore, but it inhibited Chlorella cell fission (liberation of autospores). The addition of Ca²⁺ to cells cultivated in the absence of Ca²⁺ resulted in an increase in n cell population, leading to the resumption of C. sorokiniana growth. These findings suggest that Ca²⁺ plays a crucial role in the fission process in Chlorella.

• Environmental Mutagens and Carcinogens
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• v.21 no.2
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• pp.82-88
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• 2001

Recently, there has been significant progress in understanding the control process of the cell division cycle. To investigate the influence of toxic substances on the cell cycle, the effect of benzo(a)pyrene (BAP) and mitomycine C (MMC) on synchronized HeLa cells was analyzed during the cell cycle. To synchronize the HeLa cells, 10$^{6}$ cells were grown for 1 day and then treated with 1 mM hydroxyurea for 14 h. The arrested cells were then allowed to proceed through their cell cycle by removing the hydroxyurea and resupplying a fresh medium. The arrested cells in the G1/S transition then proceeded to the S phase after 4 h, the G2/M phase after 8h, and the G1 phase after 12 h, subsequent to the resupply of a fresh medium. In the untreated HeLa cells, the p34$^{cdc2}$ kinase activity, measured using a p34$^{cdc2}$ specific peptide, peaked after 8h (G2/M) and then declined after 12 h (G1). However, treatment with 30 $\mu$M BAP delayed the peak of the p34$^{cdc2}$ kinase activity. The amount of p34$^{cdc2}$ remained unchanged in the untreated, BAP-, and MMC-treated cells throughout the cell cycle. The cyclin B level peaked after 8 h in the untreated cells, yet peaked after 10-12 h in the BAP-treated cells. There was no significant change in the cyclin B level in the MMC-treated cells.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.319-327
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• 2011

This study investigated whether the addition of porcine sperm cytosolic factor (SCF) at fusion/activation affects in vitro development of porcine parthenogenetic(PA) and nuclear transfer (NT) embryos. To determine the optimum concentration of SCF, control group of oocytes was activated with 0.3M mannitol (1.0 mM $CaCl_2{\cdot}2H_2O$), other three groups of oocytes were parthenogentically activated with the fusion medium (0.1mM $CaCl_2{\cdot}2H_2O$) supplemented with 100, 200 or 300 ${\mu}$g/ml SCF, respectively. Matured oocytes were activated with two electric pulses (DC) of 1.2 kv/cm for 30 ${\mu}$sec. The activated embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. Oocytes activated in the presence of SCF showed a significantly higher blastocyst rate than control (p<0.05). Apoptosis rate was significantly lower in 100 ${\mu}$g/ml SCF group than other groups (p<0.05). Cdc2 kinase activity in control and SCF treatment group of oocytes was determined using MESACUP cdc2 kinase assay kit at 1, 5, 10, 15, 30, 45 and 60 min after activation. Cdc2 kinase activity was significantly decreased (p<0.05) in SCF group than MII oocytes or control within 5 min. For NT embryo production, reconstructed oocytes were fused in the fusion medium supplemented with 0.1 mM $CaCl_2{\cdot}2H_2O$ (T1), 1.0 mM $CaCl_2{\cdot}2H_2O$ (T2) and 0.1 mM $CaCl_2{\cdot}2H_2O$ with 100 ${\mu}$g/ml SCF (T3). Fused embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. Developmental rate to blastocyst stage was significantly higher in T3 than other groups (23.0% vs. 13.5 to 15.2%) (p<0.05). Apoptosis rate was significantly lower in T3 than T1 or T2 (p<0.05). The relative abundance of Bax-${\alpha}$/Bcl-xl was significantly lower in in vivo or SCF group than that of control (p<0.05). Moreover, the expression of p53 and caspase3 mRNA was significantly lower in in vivo or SCF group than that of control (p<0.05). These results indicate that the addition of SCF at fusion/activation might improve in vitro development of porcine NT embryos through regulating cdc2 kinase level and expression of apoptosis related genes.

• Journal of Ginseng Research
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• v.23 no.3 s.55
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• pp.190-197
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• 1999

In this study, the molecular mechanism of the growth inhibitory effect of panaxynol was investigated in a human malignant melanoma cell line, SK-MEL-l. In the cell cycle analysis, panaxynol arrested cell cycle progression of SK-MEL-I at the G1 phase. Immunoblot analysis demonstrated that panaxynol increased $p21^{WAF1}$ and decreased cdc2 expression. Protein levels of pl6, p27, E2F-1, Rb, and p53 were not changed. Thus, the changes in expression levels of $p21^{WAF1}$ and cdc2 apparently mediate the cell cycle arrest caused by panaxynol. In addition, cycloheximide (CHX) partially reversed the growth inhibition by panaxynol, which suggested that new protein synthesis was required. On the other hand, LLnL, a proteasome inhibitor, increased antiproliferative effect of panaxynol. This may be due to stabilization of the protein(s) responsible for the growth inhibition such as $p21^{WAF1}$. In summary, these results demonstrate that panaxynol inhibits proliferation of SK-MEL-I by inducing cell cycle arrest at the G1 phase and the inhibitory effect is mediated by the increased level of $p21^{WAF1}$ as well as decreased cdc2 expression.

• Journal of Korean Medicine Rehabilitation
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• v.18 no.3
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• pp.19-39
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• 2008

Objectives : It has been reported that CG was effective in decreasing injury to neural tissues. To investigate neural responses in the injured spinal cord, an extract of CG was examined to determine its effect on neural responses in the injured spinal cords of rats. Methods : After CG treatment was applied to the spinal cord of rats given a contusion injury, the re-growth responses of injured neural tissues and corticospinal tract axons was observed by measuring the number of GAP-43, Cdc2, and phospho-Erk1/2 proteins, CST axons, GFAP-stained astrocytes, and Glial scarring in the injured spinal cord. Results : Levels of GAP-43, Cdc2, and phospho-Erk1/2 proteins were found to have increased in the injured spinal cord region. The number of GFAP-stained astrocytes also increased within and around the injury cavity. Glial scarring, which was identified by CSPG immunofluorescence staining, was reduced by CG treatment. Anterograde tracing by Dil dye showed that the elongation of the CST axons in the dorso-medial white matter area was almost completely prevented at the injury site. Collateral sprouting was observed in the spinal cord rostrally close to the injury site, and CG treatment further increased axonal arborization in the corresponding region. In vivo migration of CST axons and astrocytes using an implanted polymer tube system showed more of an increase in enhanced migration of axons and astrocytes in CG-treated group compared to the injury control group. Conclusions : These results suggest that CG activated neural responses - including astrocyte migration - and promotes axonal regenerative activity in the injured spinal cord area.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.6
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• pp.2965-2972
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• 2016

Background: China carries a heavy burden of cervical cancer (CC) and substantial disparities exist across regions within the country. In order to reduce regional disparities in CC, the government of China launched the National Cervical Cancer Screening Program in Rural Areas (NCCSPRA) in 2009. Critical to the success of the program are the health care workers who play a pivotal role in preventing and managing CC by encouraging and motivating women to use screening services and by providing identification and treatment services. This study aimed to assess cervical cancer knowledge among these health care workers at the county level in maternal and child health (MCH) hospitals across different socio-economic regions of China. Materials and Methods: A cross-sectional survey was conducted and self-administered questionnaires were sent to all health care workers (a total of 66) providing cervical cancer screening services in 6 county level MCH hospitals in Liaoning, Hubei and Shaanxi provinces, representing eastern, central and western regions of China; 64 (97.0%, 64/66) of the workers responded. ANOVA and Chi-square test were used to compare the knowledge rate and scores in subgroups. Results: The knowledge level of the respondents was generally low. The overall combined knowledge rate was 46.9%. The knowledge rates for risk factors, prevention, clinical symptoms, screening and diagnostic tests and understanding of positive results were 31.3%, 37.5%, 18.1%, 56.3% and 84.4%, respectively. Statistically significant differences in scores or rates of CC knowledge were seen across the different regions. The total and sectional scores in the less developed regions were statistically significantly lower than in the other regions. Conclusions: The majority of the health care workers who provide CC screening service in NCCSPRA at county level MCH hospitals do not have adequately equipped with knowledge about CC. Given the importance of knowledge to the program's success in reducing CC burden in rural women in China, efforts are needed to improve the knowledge of health care workers, especially in less developed regions.

• Journal of Periodontal and Implant Science
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• v.49 no.3
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• pp.138-147
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• 2019

Purpose: Several studies have shown that the oral cavity is a secondary location for Helicobacter pylori colonization and that H. pylori is associated with the severity of periodontitis. This study investigated whether H. pylori had an effect on the periodontium. We established an invasion model of a standard strain of H. pylori in human periodontal ligament fibroblasts (hPDLFs), and evaluated the effects of H. pylori on cell proliferation and cell cycle progression. Methods: Different concentrations of H. pylori were used to infect hPDLFs, with 6 hours of co-culture. The multiplicity of infection in the low- and high-concentration groups was 10:1 and 100:1, respectively. The Cell Counting Kit-8 method and Ki-67 immunofluorescence were used to detect cell proliferation. Flow cytometry, quantitative real-time polymerase chain reaction, and western blots were used to detect cell cycle progression. In the high-concentration group, the invasion of H. pylori was observed by transmission electron microscopy. Results: It was found that H. pylori invaded the fibroblasts, with cytoplasmic localization. Analyses of cell proliferation and flow cytometry showed that H. pylori inhibited the proliferation of periodontal fibroblasts by causing G2 phase arrest. The inhibition of proliferation and G2 phase arrest were more obvious in the high-concentration group. In the low-concentration group, the G2 phase regulatory factors cyclin dependent kinase 1 (CDK1) and cell division cycle 25C (Cdc25C) were upregulated, while cyclin B1 was inhibited. However, in the high-concentration group, cyclin B1 was upregulated and CDK1 was inhibited. Furthermore, the deactivated states of tyrosine phosphorylation of CDK1 (CDK1-Y15) and serine phosphorylation of Cdc25C (Cdc25C-S216) were upregulated after H. pylori infection. Conclusions: In our model, H. pylori inhibited the proliferation of hPDLFs and exerted an invasive effect, causing G2 phase arrest via the Cdc25C/CDK1/cyclin B1 signaling cascade. Its inhibitory effect on proliferation was stronger in the high-concentration group.

• Journal of Embryo Transfer
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• v.16 no.3
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• pp.223-231
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• 2001

The success of nuclear transplantation with mammalian oocytes depends critically on the potential of oocytes activation, which mainly caused to prevent the re-accumulation of maturation promoting factor (MPF). This study was conducted to compare the effect of combined treatment of lonomycin with a Hl-histone kinase inhibitor (dimethylaminopurine, DMAP) or cdc2 kinase inhibitor (sodium pyrophosphate, SPP) on activation of bovine oocytes. In vitro matured bovine oocytes with the first polar body (PB) and dense cytoplasm were assigned to 3 experimental groups. For activation treatment, oocytcs were exposed to 5 $\mu$M lonomycin for 5 min (Group 1), and followed by 1.9 mM dimethylaminopurine (DMAP) for 3 h (Group 2) or followed by 2 mM sodium pyrophosphate (SPP) for 3 h (Group 3). The activation effects in the three treatments and the control group (untreated) were judged by the extrusion of the second PB and formation of a pronucleus (PN). Differences among groups were analysed using one-way ANOVA after arc-sine transformation of proportional data. All three treatments led to high activation rates (90% to 95%), with significant difference from the control. However, the extrusion of the second PB and the rate of PN formation differed remarkably among treatments. In Group I and 3, about 95% of the oocytes had extruded the second polar body, but one PN had formed in a higher proportion of oocytes in Group 3 than in Group 1 (90% vs. 5%). In experiment 2, the rates of cleavage and development into blastocysts in Group 1 were significantly lower than those of Group 2 and 3 (8.7% and 0% vs. 50.5% and 11.6%, and 44.6% and 7.2%, respectively, P<0.05). In experiment 3, ~80% of parthenotes in Group 1 were developed with haploid chromosomal sets. However, when ionomycin was followed immediately by DMAP (Group 2). only 20% of parthenotes were haploid. In Group 3, combined treatment with ionomycin and SPP, the appearance of abnormal chromosomal tracts was significantly (P〈0.05) reduced and the proportion of haploid parthenotes was increased to 85% (17/20) than in Group 2. These results demonstrate that SPP acted as a cdc2 kinase inhibitor and formed the haploidy in oocyte activation. Thus, the present study suggests that cdc2 kinase inhibitor, such as sodium pyrophosphate, may have an effective role in oocyte activation for the production of cloned embryos/animals by nuclear transplantation.