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Secretion and Expression of Matrix Metalloproteinase-2 and 9 from Bone Marrow Mononuclear Cells in Myelodysplastic Syndrome and Acute Myeloid Leukemia

  • Chaudhary, Ajay K;Chaudhary, Shruti;Ghosh, Kanjaksha;Shanmukaiah, Chandrakala;Nadkarni, Anita H
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1519-1529
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    • 2016
  • Background: Matrix metalloproteinase -2 (gelatinase-A, Mr 72,000 type IV collagenase, MMP-2) and -9 (gelatinase-B, Mr 92,000 type IV collagenase, MMP-9) are key molecules that play roles in tumor growth, invasion, tissue remodeling, metastasis and stem-cell regulation by digesting extracellular matrix barriers. MMP-2 and -9 are well known to impact on solid cancer susceptibility, whereas, in hematological malignancies, a paucity of data is available to resolve the function of these regulatory molecules in bone marrow mononuclear cells (BM-MNCs) and stromal cells of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Objectives: The present study aimed to investigate mRNA expression and gelatinase A and B secretion from BM-MNCs in vitro and genotypic associations of MMP-2 (-1306 C/T; rs243865), MMP-9 (-1562 C/T; rs3918242), tissue inhibitor of metalloproteinase -1 (TIMP-1) (372T/C; rs4898, Exon 5) and TIMP-2 (-418G/C; rs8179090) in MDS and AML. Results: The study covered cases of confirmed MDS (n=50), AML (n=32) and healthy controls (n=110). MMP-9 mRNA expression revealed 2 fold increased expression in MDS-RAEB II and 2.5 fold in AML M-4 (60-70% blasts). Secretion of gelatinase-B also revealed the MMP-9 mRNA expression and ELISA data also supported these data. We noted that those patients having more blast crises presented with more secretion of MMP-9 and its mRNA expression. In contrast MMP-9 (-1562 C/T) showed significant polymorphic associations in MDS (p<0.02) and AML (p<0.02). MMP-9 mRNA expression of C/T and T/T genotypes were 1.5 and 2.5 fold increased in MDS and AML respectively. In AML, MMP-2 C/T and T/T genotypes showed 2.0 fold mRNA expression. Only MMP-9 (-1306 C/T) showed significant 4 fold (p<0.001) increased risk with chemical and x-ray exposed MDS, while tobacco and cigarette smokers have 3 fold (p<0.04) risk in AML. Conclusions: In view of our results, MMP-9 revealed synergistic secretion and expression in blast crises of MDS and AML with 'gene' polymorphic effects and is significantly associated with increased risk with tobacco, cigarette and environmental exposure. Release and secretion of these enzymes may influence hematopoietic cell behavior and may be important in the clinical point of view. It may offer valuable tools for diagnosis and prognosis, as well as possible targets for the treatments.

Topological Imitations and Reni-Mecchia-Zimmermann's Conjecture

  • Kawauchi, Akio
    • Kyungpook Mathematical Journal
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    • v.46 no.1
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    • pp.1-9
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    • 2006
  • M. Reni has shown that there are at most nine mutually inequivalent knots in the 3-sphere whose 2-fold branched covering spaces are mutually homeomorphic, hyperbolic 3-manifolds. By observing that the Z-homology sphere version of M. Reni's result still holds, M. Mecchia and B. Zimmermann showed that there are exactly nine mutually inequivalent, knots in Z-homology 3-spheres whose 2-fold branched covering spaces are mutually homeomorphic, hyperbolic 3-manifolds, and conjectured that there exist exactly nine mutually inequivalent, knots in the true 3-sphere whose 2-fold branched covering spaces are mutually homeomorphic, hyperbolic 3-manifolds. Their proof used an argument of AID imitations published in 1992. The main result of this paper is to solve their conjecture affirmatively by combining their argument with a theory of strongly AID imitations published in 1997.

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Geological Structure and Deformation History in the Gwangju area, Gyeonggi-do, Korea (경기도 광주시 일대의 지질구조와 변형사)

  • Lee Hee-Kwon;Kim Man-Kwang
    • The Journal of the Petrological Society of Korea
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    • v.14 no.2 s.40
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    • pp.108-115
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    • 2005
  • Gyeonggi metamorphic complex in the Gwangju area include banded biotite gneiss and quartzofeldspathic gneiss. Detailed structural analysis suggests that structural elements in the study area were formed by at least five phase of deformations. Penetrative compositional foliations(S1) formed in the banded gneiss during the first metamorphism and deformation (D1). After intrusion of plutonic rocks, the second deformation (D2) produced S2 foliations in the banded gneiss and quartzofeldspathic gneiss during the second metamorphism. D3 structures are represented by isoclinal folds (F3) whose axial surfaces are parallel to S3 foliations. The N-S oriented shortening (D4) was accommodated by closed upright F4 fold with about 100m of axial surface separation. F4 fold is refolded by regional F5 folding resulting in different orientation and fold style of F4 fold according to the position of F5 fold. The F4 fold with tight interlimb angle is subparallel to the axial surface (north-south) of F5 fold in the core of the F5 fold. In contrast the F4 fold trends northeast in the western limb and northwest in the eastern limb of F5 fold. The interlimb angle is larger in the limbs than that in the core of F5 fold. The trace of foliations is constrained by mainly F4 and F5 folds. Joint fanning around fold is developed in the limbs of F5 fold and bc joints are dominant in the hinge area of F5 fold. A strike-slip fault had developed in tile central part of the study area after F5 folding. The orientation of joint and foliation is rotated anticlockwise about $15^{\circ}$ by the landslide occurred during the Quaternary.

Cloning and Site-Directed Mutagenesis of Musca domestica Acetylcholinesterase for Enhancing Sensitivity to Organophosphorus and Carbamate Insecticides

  • Kim, Chung-Sei;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1760-1772
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    • 2006
  • Mature acetylcholinesterase (AChE) gene (gm, 1,836 bp) was cloned from the housefly and successfully expressed in the E. coli CodonPlus (DE3) RIL system (GM-E, 72 kDa) with a yield of 1,630 mU/g fresh cells. Using the gm, 10 kinds of mutants were constructed and expressed for enhancing sensitivity to insecticides. The sensitivity of these mutants to five kinds of organophosphate (OP) and three carbamate insecticides was investigated by measuring the apparent bimolecular inhibition constant ($k_i=k_2/K_d$). Surprisingly, the sensitivity of quadruple mutant IGFT was enhanced as much as 7-fold for acephate, 164-fold for demeton-S-methyl, 484-fold for dichlorvos, 523-fold for edifenphos, 30-fold for ethoprophos, 30-fold for benfuracarb, 404-fold for carbaryl, and 107-fold for furathiocarb, compared with that of GM-E, although the sensitivity of each single point mutant was slightly increased. These mutational studies indicated that (i) contradictory to Walsh et al. [39], the residue 327 is the important key residue for enhancing sensitivity as much as the residue 262, (ii) the residue 82 and additional residues of 234, 236, and 585 are also important, and (iii) sensitivity was cooperatively accelerated as the number of strategic mutations increased.

Cell Free EGFR mRNA Expression and Implications for Survival and Metastasis in Non-Small Cell Lung Cancer Cases

  • Masroor, Mirza;Mir, Rashid;Javid, Jamsheed;Prasant, Y;Imtiyaz, A;Mariyam, Z;Mohan, Anant;Ray, PC;Saxena, Alpana
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.15
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    • pp.6445-6449
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    • 2015
  • Background: NSCLC is a disease involving uncontrolled cell growth, which could result in metastases into nearby tissues beyond the lungs. Materials and Methods: The aim of the present study was to analyze the influence of epidermal growth factor receptor (EGFR) gene expression on metastasis and survival in NSCLC patients. The present case-control study included 100 cases of NSCLC patients and 100 age and sex matched controls. EGFR gene expression was analyzed by quantitative real time PCR using serum RNA. Association with NSCLC patient survival was analyzed by the Kaplan-Meier method. Results: We analyzed EGFR gene expression and observed mean increased gene expression of 13.5 fold in NSCLC patients. Values reflected overall survival of patients with a median of 15.8 months in the cases of <13 fold increased gene expression vs 6.7 months with >13 fold increased EGFR gene expression (p=0.005). Distant metastatic patients with <13 fold increased EGFR gene expression had 7.9 months of median survival time while>13 fold increased EGFR gene expression had only 5 months of median survival time (p=0.03). Non metastatic patients with <13 fold increased EGFR gene expression had 18 months of median survival time as compared to only 7.1 months with >13 fold increased expression. Conclusions: Higher cell free EGFR mRNA expression may play an important role in causing distant metastases and reducing overall survival of NSCLC patients in the Indian population.

A Study on the Development of Traditional Korean Paper Changphanji by the Bast Fiber of Paper Mulberry (닥나무의 인피섬유(靭皮纖維)를 이용(利用)한 전통(傳統) 한지장판지(韓紙壯版紙) 개발(開發)에 관(關)한 연구(硏究))

  • Cheon, Cheol
    • Journal of the Korean Wood Science and Technology
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    • v.18 no.4
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    • pp.53-64
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    • 1990
  • This study was carried out to develop the traditional korean paper changphanji from the bast fiber of paper mulberry The results obtained were as follows: 1. The variations in the basis weight were remarkable, but there were no significant adverse effects on the tensile strength and thickness. 2. The stamping have contributed to the improvement of smoothness and the control of thickness. The automatic pressing system recommended to obtain good control of thickness. 3. The average density of the traditional korean paper changphanji was 1.06g/$cm^3$. 4. The changphanji developed in this study gave a clude, but excellent toughness and warm feeling. Some fiber lumps, dark spots and bamboo, screen marks were also obserded. 5. The tensile strength of four-fold sheet changphanji is highest and those of two-fold sheet changphanji is lowest. the lengthwise tensile strength was higher than crosswise one due to the fiber. 6. The elongation of three-fold sheet changphanji was highest 7. The water absorption of all products ranged from 11.3g/$m^2$ to 17.5g/$m^2$. 8. The wight loss by abrasion was 34.0%(four-fold sheet) 31.6%(three fold sheet), 31.0% (two-fold sheet)respectively. 9. Based on the results obtained. tensile strength and elongation, the four-fold sheet was excellent. the use of the bast fiber of paper mulberry as a raw material is attractive point which can not obtained from the commercial changphanji made from other raw materials.

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Influence of Morinda citrifolia (Noni) on Expression of DNA Repair Genes in Cervical Cancer Cells

  • Gupta, Rakesh Kumar;Bajpai, Deepti;Singh, Neeta
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3457-3461
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    • 2015
  • Background: Previous studies have suggested that Morinda citrifolia (Noni) has potential to reduce cancer risk. Objective: The purpose of this study was to investigate the effect of Noni, cisplatin, and their combination on DNA repair genes in the SiHa cervical cancer cell line. Materials and Methods: SiHa cells were cultured and treated with 10% Noni, $10{\mu}g/dl$ cisplatin or their combination for 24 hours. Post culturing, the cells were pelleted, RNA extracted, and processed for investigating DNA repair genes by real time PCR. Results: The expression of nucleotide excision repair genes ERCC1, ERCC2, and ERCC4 and base excision repair gene XRCC1 was increased 4 fold, 8.9 fold, 4 fold, and 5.5 fold, respectively, on treatment with Noni as compared to untreated controls (p<0.05). In contrast, expression was found to be decreased 22 fold, 13 fold, 16 fold, and 23 fold on treatment with cisplatin (p<0.05). However, the combination of Noni and cisplatin led to an increase of 2 fold, 1.6 fold, 3 fold, 1.2 fold, respectively (p<0.05). Conclusions: Noni enhanced the expression of DNA repair genes by itself and in combination with cisplatin. However, high expression of DNA repair genes at mRNA level only signifies efficient DNA transcription of the above mentioned genes; further investigations are needed to evaluate the DNA repair protein expression.

Cholesterol Biosynthesis from Lanosterol: Development of a Novel Assay Method, Characterization, and Solubilization of Rat Hepatic Microsomal Sterol Δ7-Reductase

  • Lee, Joon-No;Paik, Young-Ki
    • BMB Reports
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    • v.30 no.5
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    • pp.370-377
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    • 1997
  • A novel assay method is described for rapid quantitation of reaction rate of sterol ${\Delta}^7$-reductase (${\Delta}^7$-SR) which catalyzes reduction of the ${\Delta}^7$-double bond of sterols. Of six different organ tissues-liver, small intestine, brain, lung, kidney, and testis-. ${\Delta}^7$-SR activity was detected only in liver (2.30 nmol/min/mg protein) and testis (0.11 nmol/min/mg protein). Using a newly developed method which employs diet-induced enzyme proteins and ergosterol as substrate, we assessed both kinetics ($K_m=210\;{\mu}M$, $V_{max}=1.93\;nmol/min/mg$) and inhibition of the rat hepatic ${\Delta}^7$-SR against well-studied cholesterol lowering agents such as triparanol ($IC_{50}=16\;{\mu}M$). 3-$\beta$-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A) ($IC_{50}=5.2\;{\mu}M$), and trans-1.4-bis(2-chlorobenzylaminomethyl)cyclohexane dihydrochloride (AY-9944) ($IC_{50}=0.25\;{\mu}M$). Of the three well-known AY-9944-sensitive cholesterogenic enzymes (i.e., ${\Delta}^7$-SR, sterol ${\Delta}^8$-isomerase, and sterol ${\Delta}^14$-reductase). ${\Delta}^7$-SR was found to be the most sensitive enzyme with a noncompetitive inhibition of this compound ($K_i=0.109\;{\mu}M$). Substrate specificity studies of the microsomal ${\Delta}^7$-SR indicate that the relative reaction rate for 7-dehydrocholesterol and ergosterol are 5.6-fold and 1.6-fold higher than that for lathosterol. ${\Delta}^7$-SR activity was also modulated by feeding rats a diet supplemented with 0.5% ergosterol (>2.6-fold) in addition to 5.0% cholestyramine plus 0.1% lovastatin ($\simeq$5.0-fold). Finally, microsomal ${\Delta}^7$-SR was solubilized by 1.5% 3-[3-(cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and enriched on PEG (0~10%) precipitation, which should be suitable for further purification of the enzyme.

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Purification of Alginate Lyase from Streptomyces violaceoruber and the Growth Activity of Intestinal Bacteria by Degree of Polymerization of Alginate Hydrolysates (Streptomyces violaceoruber 유래 Alginate Lyase의 정제 및 Sodium Alginate 가수분해 올리고당의 중합도별 Bifidobacterium spp.과 Lactobacillus spp.에 대한 생육활성)

  • Yoon, Min;Park, Young-Seo;Park, Gwi-Gun
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.103-109
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    • 2017
  • Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/G-oligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp.

Selectivity of Oxomemazine for the $M_1$ Muscarinic Receptors

  • Lee, Shin-Woong;Woo, Chang-Woo;Kim, Jeung-Gu
    • Archives of Pharmacal Research
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    • v.17 no.6
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    • pp.443-451
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    • 1994
  • The binding characteristics of pirenzepine and oxomemazine to muscarinic receptor were studied to evaluate the selectivity of oxomemazine for the muscarinic receptor subtypes in rat cerebral microsomes. Equililbrium dissociation constant $(K_D){\;}of{\;}(-)[^3H]$quinuclidinyl benzilate$([^3H)QNB)$ determined from saturation isotherms was 64-pM. Analysis of the pirenzepine inghibition curve of [$^3H$]QNB binding to cerebral microsome indicatd the presence of two receptor subtypes with high $(K_1 = 16 nM, M_1 receptor)$two receptor subypes with about 20-fold difference in the affinity for high $(k_1 = 84nM, {\;} O_H receptor){\;} and {\;}low{\;} (K_1{\;} ={\;} 1.65\muM, {\;} O_L receptor$) affinity sites. The percentage populations of $M_1{\;} and M_3$, /TEX> receptors to the total receptors were 61 : 39, and those of $O_H{\;} and{\;} O_L$ receptors 39 : 61, resepectively. Both pirenzepine and oxomemazine increaed the $K_D$ value for $[^3H]QNB$ without affecting the binding site concentrations and Hii coefficient for the $[^3H]QNB$ without affecting the binding site concentractions and Hill coefficient for the [$^{3}$H]QNB binding. Oxomemazine had a 10-fold higher affinity at $M_1$ receptors than at $M_3$ receptors, and pirenzepine a 8-fold higher affinity at $O_H$ receptors were of $O_H$ receptors and 71% of $M_3$ receptors. However, $M_3$for oxomemazine and $O_H$for pirenzepine were composed of a uniform population. These results suggest that oxomemazine could be classified as a selective drug for $M_1$ receptors and also demonstrate that rat cerebral microsomes contain three different subtypes of $M_1{\;} M_3$ and the other site which is different from $M_1, {\;} M_2$, /TEX> receptors.

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