Protein phosphatase manganese dependent 1D (PPM1D), a Ser/Thr protein phosphatise, play major role in the cancer tumorigenesis of various tumors including neuroblastoma, pancreatic adenocarcinoma, medulloblastoma, breast cancer, prostate cancer and ovarian cancer. Hence, analysis on the structural features required for the formation of PPM1D-inhibitor complex becomes essential. In this study, we have performed molecular docking of SL-175 and -176 and protein-protein docking of CDC5L with PPM1D. On analysing the docked complexes, we have identified the important residues involved in the formation of protein-ligand complex. Research concentrating on these residues could be helpful in understanding the pathophysiology of various tumors related to PPM1D.
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.
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.
Park Tae Yeol;Park Cheol;Yoon Hwa Jung;Choi Yung Hyun;Ko Woo Shin
Journal of Physiology & Pathology in Korean Medicine
/
v.18
no.5
/
pp.1449-1455
/
2004
Bufonis venenum (dried toad venom; Chinese name, Chan su) is a traditional Chinese medicine obtained from the skin venom gland of the toad. It has long been used in treating arrhythmia and other heart diseases in China and other Asian countries. Additionally, Bufonis venenum has been reported to selectively inhibit the growth of various lines of human cancer cells. In the present study, it was examined the effects of extract of Bufonis venenum (EBV) on the growth of human bladder carcinoma cell line T24 in order to investigate the anti-proliferative mechanism and induction of apoptosis by EBV. Treatment of T24 cells to EBV resulted in the growth inhibition, morphological change and induction of apoptotic cell death in a dose-dependent manner. Flow cytometric analysis revealed that EBV treatment caused G2/M phase arrest of the cell cycle and down-regulation of cyclin A, cyclin B1 and Cdc2, which was associated with a marked up-regulation of cyclin-dependent kinases (Cdks) inhibitor p21 (WAF1/CIP1) in a p53-independent manner. The Cdc25C expression was also significantly inhibited by EBV treatment, however Wee1 kinase expression was not affected. The induction of apoptotic cell death by EBV was connected with down-regulation of anti-apoptotic Bcl-XS/L expression without alteration pro-apoptotic Bax expression. Taken together, these findings suggest that EBV may be a potential chemotherapeutic agent for the control of human bladder carcinorma cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of EBV.
This study studied samples taken off surfaces at three sites (Unit chairs, light handles, cuspidors) of 19 dental hospitals and 28 clinics located in Gyeonggi-do and Incheon, South Korea. The bacterial contamination levels of surfaces were $44.82{\times}10^3CFU/mL$ in cuspidors, higher than in unit chairs ($5.47{\times}10^3CFU/mL$) and light handles ($16.28{\times}10^3CFU/mL$). The values were statistically higher at dental hospitals than at dental clinics, the greater number of patients being associated with the higher bacterial cell count in the cuspidors. The results of identifying the strains isolated purely from surfaces at dental clinic showed Gram positive 47.3% and Gram negative 52.7%. Among Gram positive, the most numberous bacteria were Micrococcus luteus (10.9%), Bacillus pumilus (3.6%), and Staphylococcus aureus (3.6%). Among Gram negative, the most numberous bacteria were Acinetobacter ursingii (5.5%), Brevundimonas diminuta (4.5%), Chryseobacterium (Flavo.) indologenes (CDC IIb) (4.5%), and Methylobacterium sp. (4.5%). This study measures the level of bacterial contamination and identifies the strains isolated in dental clinics. It recognizes the importance of infection control, and the results of the study may be considered as the basis for establishing specific plans for prevention of infection.
N-acetylcysteine (NAC) has been widely used as an initial mucolytic agent and is generally used as an antioxidant to help alleviate various inflammatory symptoms. NAC reduces bacterial extracellular polymeric substances (EPS) production, bacterial adhesion to the surface and strength of mature biofilm. The efficacy has been shown to inhibit proliferation of gram-positive and gram-negative bacteria. In membrane bioreactor (MBR) processes, which contain a variety of gram negative bacteria, biofilm formation has become a serious problem in stable operation. In this study, use of NAC as an inhibitor of biofilm contamination was investigated using the center for disease control (CDC) reactors with MBR sludge. Biomass reduction was confirmed with CLSM images of membrane surfaces by addition of NAC, which was more efficient as the concentration of NAC was increased to 1.5 mg/mL. NAC addition also showed decreases in EPS concentrations of the preformed biofilm, indicating that NAC was able to degrade EPS in the mature biofilm. NAC addition was also effective to inhibit biofilm formation by MBR sludge, which consisted of various microorganisms in consortia.
This study was conducted to monitor and prevent outbreak of insect pestsin dredged soil dumping area after completion of dredging construction in Masan City. Monitoring was carried out using tent trap, colored sticky trap, and CDC light trap. A total of 217,073 individuals belonging to 23 species from 10 families in 3 orders were collected. In overwintering survey using tent trap, 3 species were collected. 2 species (Leptocera fuscipennis (Haliday) and Ephydra japonica Miyagi) of them were outbreak species. In color sticky trap, more than 96% of total individuals were comprised of five species: Urolepis maritima Walker (43%), E. japonica (19%), Fucellia sp. 1 (13%), Philotelma sp. 1 (10%), and Homalometopus sp. 1 (9%). In CDC light trap, three dominant species were Homalometopus sp. 1 (91%), Glyptotendipes tokunagai Sasa (6%), and L. fuscipennis (1%), representing about 98% of the total. To prevent damage caused by outbreak of insect pests, we carried out ecological control methods such as covering the fresh soil in outbreak area, using light trap, pumping up water and so on, minimizing use of thermal fogging and insect growth regulatorwhen the insect pest population was rapidly increasing.
Licochalcone (LC), isolated from the roots of Glycyrrhiza inflata has multiple pharmacological effects including anti-inflammatory and anti-tumor activities. To date, Licochalcone C (LCC) has induced apoptosis and inhibited cell proliferation in oral and bladder cancer cells, but lung cancer has not yet been studied. In addition, no study reported LCC-induced autophagy in cancer until now. The present study was designed to investigate the effect of LCC on gefitinib-sensitive and -resistant lung cancer cells and elucidate the mechanism of its action. The 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay data showed that LCC significantly inhibited cell viability in non-small cell lung cancer (NSCLC) HCC827 (gefitinib-sensitive) and HCC827GR (gefitinib-resistant) cell lines. Interestingly, Annexin V/7-aminoactinomycin D double staining and cell cycle analysis showed an apoptosis rate within about 20% at the highest concentration of LCC. LCC induced G2/M arrest by reducing the expression of the cell cycle G2/M related proteins cyclin B1 and cdc2 in NSCLC cell lines. Treatment of LCC also induced autophagy by increasing the expression of the autophagy marker protein microtubule-associated protein 1 light chain 3 (LC3) and the protein autophagy-related gene 5 involved in the autophagy process. In addition, LCC increased the production of reactive oxygen species (ROS), and the cell viability was partially restored by treatment with the ROS inhibitor N-acetyl-L-cysteine. In western blotting analysis, the expression of cdc2 was increased and LC3 was decreased by the simultaneous treatment of NAC and LCC. These results indicate that LCC may contribute to anti-tumor effects by inducing ROS-dependent G2/M arrest and autophagy in NSCLC. In conclusion, LCC treatment may be useful as a potential therapeutic agent against NSCLC.
Objectives : The present study was performed to investigate whether acupuncture stimulation in the rats affected regeneration properties of the injured sciatic nerve. A differential effect of acupuncture stimulation on the one point near the spinal nerve root controlling sciatic nerve activity and the other point in the peripheral area subordinated by injured nerve was compared. Materials and Methods: Rat sciatic nerves were injured by crush, and the effects on axonal regeneration on injured sciatic nerves were evaluated by acupuncture stimulation at two different regions. In proximal acupuncture stimulation group, acupuncture stimulation was performed on Huatuo Jiaji(EX B2) points located from L5 to S1 vertebral levels to stimulate the nearest spinal nerve root that innervates sciatic nerves. In distal acupuncture stimulation group, acupuncture stimulation was performed on Zusanli(ST 36) and Weizhong(BL 40) points to stimulate at peripheral area dominated by injured sciatic nerves. Acupuncture stimulation was given every other days for 1 or 2 weeks. Sciatic nerve tissues collected from acupuncture stimulation experimental groups, injury control group, and intact animal group were used for protein analysis by Western blotting or Hoechst nuclear staining. To determine axonal regeneration, Dil fluorescence dye was injected into the sciatic nerve 0.5 cm distal to the injury site in individual animal groups and Dil-labeled cells by retrograde tracing were measured in the DRG at lumbar 5 or in the spinal cord. DRG sensory neurons prepared from individual animal groups were used to measure the extent of neurite outgrowth and for immunofluorescence staining with anti-GAP-43 antibody. Results : Animal groups given proximal or distal acupuncture stimulation showed upregulation of GAP-43 and Cdc2 protein levels in the sciatic nerve at 7 days after injury. Cdk2 protein levels were strongly induced by nerve injury, but did not show changes by acupuncture stimulation. Phospho-Erk1/2 protein levels were elevated by acupuncture stimulation above those present in the injury control animals. These increase in regeneration-associated protein levels appeared to be related with increase cell proliferation in the injured sciatic nerves. Hoechst 33258 staining of sciatic nerve tissue to visualize nuclei of individual cells showed increased Schwann cell number in the distal portion of the injured nerve 7 and 14 days after injury and further increases by acupuncture stimulation particularly at the proximal position. Measurement of axonal regeneration by retrograde tracing showed significantly increased Dil-labeled cells in proximal acupuncture stimulation group compared to distal acupuncture stimulation group and injury control group. Finally, an evaluation of axonal regeneration by retrograde tracing showed increased number of Dil labeled cells in the DRG at lumbar 5 or in the ventral horn of the spinal cord at lower thoracic level at 7 days after nerve injury. Conclusions : The present data show that the proximal acupuncture stimulation at Huatuo Jiaji(EX B2) points governing injured sciatic nerves was more effective for axonal regeneration than the distal acupuncture stimulation. Further studies on functional recovery or associated molecular mechanisms should be critical for developing animal models and clinical applications.
The purpose of this research is to assess hematological and biochemical status and the prevalence of iron deficiency of pregnant women by gestational age to provide the primary data about iron nutritional status of pregnant women. Pregnant women visiting public health centers in Ulsan participated in study and were divided into 3 trimester by last menstrual period(LMP). Hemoglobin (Hgb), hematocrit(Hct)and mean corpuscular volume(MCV) among iron status indices were not statistically different from normal distribution, however total iron binding capacity(TIBC) and serum ferritin were skewed to left and serum iron and transferrin saturation(TS) were skewed to right. Hgb was positively correlated with Hct(r=0.93, p<0.001) but TIBC was negatively correlated with all indices. Serum ferritin was also correlated with all indices, especially in 3rd trimester but not reached to 1st trimester level. Mean corpuscular hemoglobin(MCH), mean corpuscular hemoglobin concentration(MCHC), Red cell distribution width(RDW), serum iron and TS were not significantly different by trimester, however when serum serum iron was adjusted with hematocrit to correct the hemodilution, it significantly decreased in 2nd trimester. MCV increased in 2nd trimester and was maintained until late pregnancy, TIBC continued to increase throughout the trimester. The prevalence of anemic by CDC(Centers for Disease Control) Hgb criteria(Hgb <11.0g/dl in 1st and 3nd trimester, Hgb<10.5g/dl in 2nd trimester) was 2.8% in 1st trimester, 22.5% in 2nd trimester, 27.1% in 3rd trimester and was similar with prevalence by CDC Hct criteria(Hct < 33% in 1st and 3rd, Hct < 32% in 2nd). The prevalence of anemic of total subjects was 32.7% by WHO criteria(Hgb < 11.0g/dl). Although almost iron status indices increased in 3rd trimester, the prevalence of anemia by different criteria of all indices increased throughout the trimester, so iron nutritional status was considered as serious during late pregnancy. However, since factors other than iron deficiency, such as infection, infection, inflammation, other nutrient deficiency may also play a significant role, to differentiate the anemia due to mainly iron deficiency from the anemia due to other factors, serum ferritin is among the more useful indices in distinguishing the two conditions because it is depressed only in iron deficiency. Hgb<11.0g/dl and serum ferritin<12.0ug/L as the criteria of iron deficiency was suggested by CDC. 17.8% of all subjects were classified as iron deficient anemia, 14.9% as anemic from other reasons, 21.2% as iron deficiency any only 46.2% were in normal iron status.
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