• Animal cells and systems
• /
• v.10 no.1
• /
• pp.1-6
• /
• 2006

Previous studies showed that Cdc7 kinase of Schizosaccharomyces pombe phosphorylated the minichromosome maintenance (Mcm) complex efficiently in the presence of spMcm10 protein. The biochemical properties of the phosphorylated Mcm complexes were examined to understand the activation mechanism of the Mcm complex by Cdc7 kinase. The phosphorylation of Mcm complex in the presence of spMcm10 by Cdc7 kinase did not affect the stability of the Mcm complex containing all six subunits, and the changes in the sedimentation properties were not observed after the phosphorylation. The reconstitution of the Mcm complex using the purified proteins showed that the phosphorylation of Mcm2 proteins did not affect the interactions between Mcm proteins. The phosphorylation of the Mcm2-7 complex at the same condition also did not activate the other biochemical activities such as DNA helicase and single stranded (ss) DNA binding activities. On the other hand, spMcm10 protein that was used for the stimulation of Mcm phosphorylation showed single stranded DNA binding activity, and inhibited the DNA helicase activity of the Mcm4/6/7 complex. These inhibitory effects were reduced by the addition of Cdc7 kinase, suggesting that the phosphorylation by Cdc7 kinase decreased the interactions between spMcm10 and the Mcm complex. Taken together, these results suggested that the phosphorylation by Cdc7 kinase alone is not sufficient for the remodeling and the activation of the Mcm complex, and the additional factors or the phosphorylations might be required for the activation of the Mcm complex.

• BMB Reports
• /
• v.40 no.5
• /
• pp.805-813
• /
• 2007

Although MCM2 is obviously important for the initiation of eukaryotic DNA replication, its role in $O_2$ dependent regulation of replicon initiation is poorly understood. In this report, I analysed the changes of MCM2 during the transition from hypoxically suppressed replicon initiation to the burst of initiation triggered by reoxygenation in T24 cells. A high level of chromatin bound and nucleosolic MCM2 was found under the hypoxic replicon arrest. In contrast low cytosolic MCM2 was noticed. Recovery of $O_2$ induced phosphorylation and diminution of chromatin bound MCM2, whereas cytosolic MCM2 increased. The level of chromatin bound Cdc7 did not change significantly upon reoxygenation. However, after reoxygenation, significant phosphorylation of Cdc7 and an increase of coimmunoprecipitation with its substrate (MCM2) were observed. This provides a hint that reoxygenation may promote the kinase activity of Cdc7. These changes might be the critical factors in $O_2$ dependent regulation of replicon initiation. Moreover, phosphorylation of Cdc7 by Cdk2 can be observed in vitro, but seems to fail to regulate the level of chromatin bound Cdc7 as well as the changes of MCM2 in response to reoxygenation of hypoxically suppressed cells.

• Journal of Haehwa Medicine
• /
• v.14 no.1
• /
• pp.201-211
• /
• 2005

Cdc2 kinase is a prototypical cyclin-dependent kinase critical for G2 to M phase cell cycle transition. Yet, its function in the nervous system is largely unknown. Here, we investigated possible role of Cdc2 in axonal regeneration using sciatic nerve system in rat. Cdc2 protein levels and activity were increased in the injured sciatic nerves 3 and 7 days after crush injury and then decreased to basal level 14 days later. Administration of Cdc2 kinase inhibitor roscovitine in vivo at the time of crush injury significantly inhibited axonal regeneration when regrowing axons were analyzed using retrograde tracers. Cdc2 protein levels in cultured Schwann cells which were prepared from sciatic nerves 7 days after crush injury were much higher compared with those from uninjured sciatic nerves, suggesting that Cdc2 protein expression was primarily induced in the Schwann cells. To further investigate Cdc2 function in Schwann cell, we examined changes in cultured Schwann cell proliferation and migration in culture system. Both the number of proliferating Schwann cells and the extent of neurite outgrowth from co-cultured DRG neurons were significantly decreased by Cdc2 inhibitor roscovitine treatment in DRG culture which was prepared from animals with sciatic nerve injury for 7 days. Also, Schwann cell migration in the injured sciatic nerve explant was significantly inhibited by roscovitine treatment. Taken together, the present data suggest that Cdc2 may be involved in peripheral nerve regeneration via Schwann cell proliferation and migration.

• Journal of Korean Dental Hygiene Science
• /
• v.6 no.1
• /
• pp.27-36
• /
• 2023

Background: The purpose of the study was to investigate the level of infection control and prevention awareness among dental practitioners in Korea based on the infection control and prevention guidelines of the Centers for Disease Control and Prevention (CDC) in the United States. Methods: A survey was created on 'Standard and Education on CDC Infection Control Standards', 'Matters related to CDC Infection Control Prevention', and 'Characteristics of Research Subjects and Infection Control Characteristics' based on the CDC Dental Infection Control List.' A total of 222 surveys were conducted and used for frequency and cross-tabulation analyses. Results: Most research participants worked at university or general hospitals, and 93.7% had received infection prevention education within the past year. The average awareness of the CDC dental infection control standards and education was 77.2 %, which was higher than previous research results. Preventive awareness was 71.5% on average, and there was a statistically significant difference in preventive awareness between the certified and non-certified evaluation groups (p<0.001). Conclusion: The participants of this study showed a higher awareness of infection control standards, education, and prevention than those in previous studies. However, this was insufficient compared with the CDC dental infection control standard prevention guidelines. Therefore, government agencies and related organizations must establish systematic infection control systems.

• The Korean Journal of Physiology and Pharmacology
• /
• v.27 no.5
• /
• pp.427-436
• /
• 2023

Mitotic arrest deficient 2 like 2 (Mad2L2, also known as Mad2B), the human homologue of the yeast Rev7 protein, is a regulatory subunit of DNA polymerase ζ that shares high sequence homology with Mad2, the mitotic checkpoint protein. Previously, we demonstrated the involvement of Mad2B in the cisplatin-induced DNA damage response. In this study, we extend our findings to show that Mad2B is recruited to sites of DNA damage in human cancer cells in response to cisplatin treatment. We found that in undamaged cells, Mad2B exists in a complex with Polζ-Rev1 and the APC/C subunit Cdc27. Following cisplatin-induced DNA damage, we observed an increase in the recruitment of Mad2B and Cdc20 (the activators of the APC/C), to the complex. The involvement of Mad2B-Cdc20-APC/C during DNA damage has not been reported before and suggests that the APC/C is activated following cisplatin-induced DNA damage. Using an in vitro ubiquitination assay, our data confirmed Mad2B-dependent activation of APC/C in cisplatin-treated cells. Mad2B may act as an accelerator for APC/C activation during DNA damage response. Our data strongly suggest a role for Mad2B-APC/C-Cdc20 in the ubiquitination of proteins involved in the DNA damage response.

• Journal of Life Science
• /
• v.7 no.2
• /
• pp.79-87
• /
• 1997

To determine the cause of Vibrio septicemia by understanding the characteristics endotoxin from Vibrio vulnificus, lethal dose, heat resistance and vascular permeability enhancing activity were svaluated using vegestative cell and cell homogenate and the result is as follows: 1. Vibrio vulnificus CDC B3547 of patient origin did not exihibit any significant difference in toxicity compared to Vibrio vulnificus B57 of enviroment origin. 2. Strong toxicity was observed when viable cell count of Vibrio vulnificus CDC B3547 was more than 10$^{7}$/ml. 3. Toxicity of cell homogenate was completely inactivited upon geating at 80$^{\circ}$C for 20min. 4. Cell homogenate did not show hemolyic activity but was acknowleged to have cytotoxicity. 5. Major lethal toxin against mouse was existed in Vibrio vulnificus CDC B3547; however, separation of LPS and LPS-protein complex was not successful using the current technique.

• Journal of Korean Medical Science
• /
• v.33 no.47
• /
• pp.303.1-303.10
• /
• 2018

Background: Cell division cycle 6 (CDC6) is an essential regulator of DNA replication and plays important roles in the activation and maintenance of the checkpoint mechanisms in the cell cycle. CDC6 has been associated with oncogenic activities in human cancers; however, the clinical significance of CDC6 in prostate cancer (PCa) remains unclear. Therefore, we investigated whether the CDC6 mRNA expression level is a diagnostic and prognostic marker in PCa. Methods: The study subjects included 121 PCa patients and 66 age-matched benign prostatic hyperplasia (BPH) patients. CDC6 expression was evaluated using real-time polymerase chain reaction and immunohistochemical (IH) staining, and then compared according to the clinicopathological characteristics of PCa. Results: CDC6 mRNA expression was significantly higher in PCa tissues than in BPH control tissues (P = 0.005). In addition, CDC6 expression was significantly higher in patients with elevated prostate-specific antigen (PSA) levels (> 20 ng/mL), a high Gleason score, and advanced stage than in those with low PSA levels, a low Gleason score, and earlier stage, respectively. Multivariate logistic regression analysis showed that high expression of CDC6 was significantly associated with advanced stage (${\geq}T3b$) (odds ratio [OR], 3.005; confidence interval [CI], 1.212-7.450; P = 0.018) and metastasis (OR, 4.192; CI, 1.079-16.286; P = 0.038). Intense IH staining for CDC6 was significantly associated with a high Gleason score and advanced tumor stage including lymph node metastasis stage (linear-by-linear association, P = 0.044 and P = 0.003, respectively). Conclusion: CDC6 expression is associated with aggressive clinicopathological characteristics in PCa. CDC6 may be a potential diagnostic and prognostic marker in PCa patients.

• BMB Reports
• /
• v.36 no.4
• /
• pp.421-425
• /
• 2003

The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non-radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially-available generic anti-phosphothreonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the $K_m$ for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• v.15 no.1
• /
• pp.29-37
• /
• 2012

Purpose: To examine the prevalence of $Clostridium$ $difficile$ ($C.$ $difficile$) colonization (CDC) and potential neonatal determinants of CDC in hospitalized preterm infants. Methods: Fecal samples were serially collected within 72 h after birth and at 1, 2, and 4-6 weeks of age from preterm infants in the neonatal intensive care units (NICUs) of two different university hospitals. Total bacterial DNA was extracted from each fecal sample from 49 infants, and polymerase chain reaction (PCR) was performed with primers for the 16S gene of $C.$ $difficile$ and the toxin A and toxin B genes. The correlation between the results of $C.$ $difficile$ PCR assays and the clinical characteristics of the infants was analyzed. Results: The prevalence rates of CDC were 34.7, 37.2, 41.3, and 53.1% within 72 h after birth and at 1, 2, and 4.6 weeks of age, respectively. The toxin positivity rate was significantly higher in the infants with persistent CDC than in those with transient CDC (8/12 [66.7%] vs. 6/25 [24.5%] ($p$=0.001). Among the various neonatal factors, only the feeding method during the first week after birth was significantly associated with persistent CDC. Exclusive breast-milk feeding (EBMF) significantly decreased the risk of persistent CDC compared to formula or mixed feeding (adjusted odds ratio: 0.133, 95% confidence interval: 0.02-0.898, $p$=0.038). Conclusion: The prevalence of CDC increased with the duration of hospitalization in preterm infants in the NICU. EBMF during the first week after birth in hospitalized preterm infants may protect against persistent CDC.

• Journal of Veterinary Clinics
• /
• v.22 no.4
• /
• pp.322-327
• /
• 2005

DLA class I complement-dependent cytotoxicity (CDC) cross-match method was established to control hyperacute rejections in organ transplantation. The aim of the present study is to investigate if DLA class 1 CDC corssmatch method is effective to prevent hyperacute rejections in canine renal allografts. Seven mongrel dogs of similar age and weight were used. Erythrocyte crossmatch was first performed and only the negatives were used. Among the same blood types, CDC cross-match was performed. Anti-dog serum, Hank's balanced salt solution(HBSS), and tile self-serum was used as a positive-, a negative-, and all auto-control respectively. After the reaction with class I complement, it was stained with eosin and scored by international cytotoxicity scoring system under inverted phase microscope. According to these results, kidneys oi CDC negatives among same blood types were cross-transplanted to observe the incidence of hyperacute rejections. One of four 1.2 B blood type dogs had autoantibodies. here were negative CDC results among each blood type, and also there were negative results between different blood types. Two pairs with the same blood types and negative CDC results underwent allo-transplantation each other. There were no hyperacute rejections. DLA cross-match method studied in this experiment for canine renal allograft can be effective to prevent hyperacute rejections. it may be applicable for the future studies of histocompatibility testing in canine renal allografts.