• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.457-473
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• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.

• Korean Journal of Microbiology
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• v.27 no.4
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• pp.323-333
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• 1989

Plasmid DNA molecules containing strong promoter upstream from IS50L or IS50R, the two insertion sequences that flank Tn5, were constructed to amplify the synthesis of Tn5-encoded polypeptides. When proteins made by cells that contain these plasmids were analyzed on polyacrylamide gels, enhanced synthesis of IS50R polypeptides could be detected. Synthesis of this polypeptide apparently is initiated within the large open reading frame of this element. In addition, the stability of IS50L-and IS50R-encoded polypeptides was analyzed. It was found that IS50L polypeptides are relatively unstable in vivo. This instability could account for the observed inability of this element to promote transposition.

• Journal of Periodontal and Implant Science
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• v.24 no.2
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• pp.219-237
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• 1994

The ultimate goal of clinical periodontal therapy is to achieve regeneration of a healthy connective tissue reattachment. Conventional therapy including scaling, root planing, gingival curettage, gingivectomy and flap procedures of various types results primarily in repair rather than regeneration of the periodontium. In order for periodontal regeneration to occur, progenitor periodontal ligament cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. Insulin-like growth factor-I (IGF- I ) of these factors appear to have an important role in periodontal wound healing and bone formation. The purpose of this study is to evaluate the effects of IGF- I on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were obtained from periodontal tissue explants culture of the first premolar tooth extracted for the orthodontic treatment. Cells were cultured in Dulbecco's modified Eagle medium(DMEM) with 10% fetal bovine serum. Fourth to seventh passage cells were plated in 24 well tissue culture plates and medium changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of $[^3H]-thymidine$ into DNA, Protein synthesis was determined by measurement of $[^3H]-proline$ incorporation into collagenase-digestible protein(CDP) and noncollagenous protein(NCP) according to the method of Peterkofsky and Diegelmann (1971), And alkaline phosphatase activity was measured as one parameter of osteoblastic differentiation. The results were as follows : The DNA synthetic activity was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I At the concentration of 10, 100ng/ml, IGF- I significantly increased the DNA synthetic activity(P<0.05) The total protein, collagen and noncollagen synthesis was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I. At the concentration of 1, 10, 100ng/ml, IGF- I significantly increased the total protein, collagen and noncollagen synthesis activity(P<0.95, P<0.001). The % of collagen was not effected according to the concentration of IGF- I. The alkaline phosphatase activity was increased in a dose-, time-dependent manner with IGF- I (10, 100ng/ml). In conclusions, the present study shows that IGF- I has a potentiality to enhance the DNA synthesis of periodontal ligament cells with including the increase of the total protein and collagen synthetic activity. The use of IGF- I to mediate biological stimulation of periodontal ligament cells shows promise for future therapeutic applications.

• Journal of Plant Biology
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• v.20 no.2
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• pp.91-101
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• 1977

Using barley seeds (Hordeum sativum Jess, var.), the influences of gibberellic acid (GA) and indole-3-acetic acid(IAA) on the amylase synthesis and that of the nucleic acid metabolism were investigated. 1. With the deembrynized barley seeds, the increase of amylase treated with a $10^{-5}M$ of GA and the decrease of amylase treated with $10^{-5}M$IAA were matched by a proportionate increase and decrease in the amount of RNA. The influence of the hormones on the RNA synthesis has appeared immediately after the treatment but on the amylase synthesis it has appeared 8 hours later. But no influence on the DNA synthesis was observed on both hormones. 2. The amylase from deembryonized barley seeds treated with GA and IAA have been fractionated by gel filteration on Sephadex G-100. The amylase components showed four fractions on both enzymes treated with GA and IAA. Fraction I(FI) was differed from fraction Ⅵ(FIV) in Km value and the effects of temperature, pH and metal ions. On the basis of their emzymatic properties, it was considered that the FI was $\beta$-amylase and FIV was $\alpha$-amylase. The influences of GA and IAA on each fractions appeared to be similar but on the amylase units per souble protein, IAA inhibited the production of amylase FIV while it promoted that of amylase FI. 3. An experiment was conducted to determine whether IAA inhibits GA-promoted amylase synthesis competitively or non-competitively. Using a Lineweaver-Burk plot, it was clear that IAA was acting in a non-competitive fashion. From this, IAA was probably not competing with GA at the same site, but it was acting at some other site which resutled in partial blocking of the action of GA on the amylase synthesis.

• The Korean Journal of Zoology
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• v.14 no.4
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• pp.199-204
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• 1971

The effect of 400 R total-body X-irradiation on the rate of deoxycytidine-2-$^14 C$(CdR-2-$^14 C$) into DNA and on the degradation of DNA has been studied in the liver, spleen and thymus of the rat. The postirradiation period can be divided into a radiation reaction period followed by a regeneration period. During the period of radiation reaction, which consists of days 1-2, markdely decreased CdR-2-$^14 C$ incorporation into DNA of each organ is observed. Rate of incorporation of labeled precursor in the thymus shows the most profound decrease, whereas those in the liver and spleen show similar decrease when expressed as percent of normal. The change in the amount of DNA as percent of normal exhibits a similar pattern in all organs, but the rate of decrease is larger in the spleen and thymus compared to that in the liver. The period of regeneration as judged by the incorporation experiment appears day 4 to 5, which consists of the second phase of the regeneration period. The second phase is highlighted by a markedly increased rate of CdR-2-$^14 C$ incorporation and by a slow and continued increase in the amount of DNA in all organs. The regeneration occurs faster in the liver and spleen than in the thymus which is the most radiosensitive of the all. The findings of the present experiments are strongly suggestive of the fact that the radiation-induced loss of spleen and thymus DNA as well as the radiation-caused inhibition in the CdR incorporation into DNA of the thymus are the important factors in the elevated levels of CdR in the urine and plasma.

• Journal of the Korean Chemical Society
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• v.54 no.6
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• pp.687-695
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• 2010

Complexes of thiocyanato(L)cobaloximes where L is urea, acetamide, semicrabazide and formamide were synthesized and characterized. The reaction of thiocyanato (L) cobaloximes (SCNCo$(DH)_2$(L)) with benzyl (aquo) cobaloxime $PhCH_2Co(DH)_2(OH_2)$ was found to produce a series of thiocyanato bridged dicobaloximes of a general formula of $PhCH_2Co(DH)_2SCNCo(DH_2)(L)$. Evidence for formulation as dicobaloximes containing thiocyanato ligand bridges was obtained from infrared data which show $20-45cm^{-1}$ increase in vCN upon formation of the dicobaloxime from the corresponding terminal thiocyanocobaloxime (SCNCo$(DH)_2$(L)). Further characterization of these two series was done on the basis of ($^1H$,$^{13}C$)NMR, LCMS and elemental analysis. Anti-microbial activity of thiocyanato(L)cobaloximes and thiocyanato bridged dicobaloximes were screened against E. Coli. The DNA-binding behaviors of both monomers and dimers were investigated by spectroscopic methods and viscosity measurements. The results indicated that the dimer complexes bind with calf-thymus DNA in an intercalative mode via the terminal benzyl ring into the base pairs of DNA. It was observed that the monomer complexes did not interact with DNA. Fluorescence spectra for the interaction between thiocyanato bridged dicobaloximes and DNA were also studied.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10433-10438
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• 2015

Background: Acute lymphoblastic leukemia (ALL) is the most common cancer diagnosed in children and represents approximately 25% of cancer diagnoses among those younger than 15 years of age. Aim and Objectives: This study investigated substitutions in the ATP synthase subunit 6 gene of mitochondrial DNA (mtDNA) as a potential diagnostic biomarker for early detection and diagnosis of acute lymphoblastic leukemia. Based on mtDNA from 23 subjects diagnosed with acute lymphoblastic leukemia, approximately 465 bp of the ATP synthase subunit 6 gene were amplified and sequenced. Results: The sequencing revealed thirty-one mutations at 14 locations in ATP synthase subunit 6 of mtDNA in the ALL subjects. All were identified as single nucleotide polymorphisms (SNPs) with a homoplasmic pattern. The mutations were distributed between males and females. Novel haplotypes were identified in this investigation: haplotype (G) was recorded in 34% in diagnosed subjects; the second haplotype was (C) with frequency of 13% in ALL subjects. Neither of these were observed in control samples. Conclusions: These haplotypes were identified for the first time in acute lymphoblastic leukemia patients. Five mutations able to change amino acid synthesis for the ATP synthase subunit 6 were associated with acute lymphoblastic leukemia. This investigation could be used to provide an overview of incidence frequency of acute lyphoblastic leukemia (ALL) in Saudi patients based on molecular events.

• Archives of Pharmacal Research
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• v.24 no.1
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• pp.55-63
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• 2001

Aucubin, an irdoid g1ucoside, was investigated to determine whether it has a stimulating effect on $\alpha$-amanitin excretion in $\alpha$-amanitin intoxicated rats, and whether there is binding activity to calf thymus DNA. High-performance liquid chromatography (HPLC) analysis of $\alpha$-amanitin in rat urine allowed quantitative measurement of the $\alpha$-amanitin concentration with a detection limit of 50${mu}g/ml$. In this system, a group treated with both $\alpha$-amanitin and aucubin showed that o(-amanitin was excreted about 1.4 times faster than in the $\alpha$-amanitin only treated group. Our previous results showed that the toxicity of $\alpha$-amanitin is due to specific inhibition of RNA polymerase activity and the resultant blockage of the synthesis of certain RNA species in the nucleus. However, no significant activity change on RNA polymerase from Hep G2 cells was observed when aucubin was treated with $\alpha$-amanitin at any concentration tested. Nevertheless, aucubigenin inhibited both DNA polymerase (IC50, 80.5${mu}g/ml$) and RNA polymerase (IC50, 135.0${mu}g/ml$) from the Hep G2 cells. The potential of both $\alpha$-amanitin and aucubin to interact with DNA were examined by spectrophotometric analysis. $\alpha$-Amanitin showed no significant binding capacity to calf thymus DNA, but aucubin was found to interact with DNA, and the apparent binding constant ($K_{app}$) and apparent number of binding sites per D7A phosphate ($B_{app}$) were 0.45$0.45{\times}$$10^4$ $M^{-1}$ and 1.25, respectively.

• Bulletin of the Korean Chemical Society
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• v.30 no.10
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• pp.2309-2317
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• 2009

Depurination, the release of purine bases from nucleic acids by hydrolysis of the N-glycosidic bond, gives rise to alterations of the cell genome. Though cells have evolved mechanisms to repair these lesions, unrepaired apurinic sites have been shown to have two biological consequences: lethality and base substitution errors. 2-Bromopropane (2-BP) is used as an intermediate in the synthesis of pharmaceuticals, dyes, and other organics. In addition, 2-BP has been used as a replacement for chloroflurocarbons and 1,1,1-trichloroethane as a cleaning solvent in electronics industry. However, 2-BP was found to cause reproductive and hematopoietic disorders in local workers exposed to it. Owing to the toxicity of 2-BP, there has been a tendency to use 1-BP as an alternative cleaning solvent to 2-BP. However, 1-BP has also been reported to be neurotoxic in rats. Though $N^7$-guanine adduct of 2-BP has been reported previously, massive depurination of the nucleosides and calf thymus DNA was observed in this study. We incubated the nucleosides (ddG, dG, guanosine, ddA, dA and adenosine) with excess amount 2-BP at the physiological condition (pH 7.4, $37\;{^{\circ}C}$), which were analyzed by HPLC and LC-MS/MS. In addition, the time and dose response relationship of depurination in nucleosides induced by 2-bromopropane at the physiological condition was investigated. Similarly, incubation of calf-thymus DNA with the excess amount 2-BP at the physiological condition was also performed. In addition, the time and dose response relationship of depurination in calf-thymus DNA induced by 2-BP at the physiological condition was investigated. Those results suggest that the toxic effect of 2-BP could be both from the depurination of nucleosides and DNA adduct formation.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.72-78
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• 1993

Since the 'promoter/enhancer region of the major immediate early (IE) ~ene of human cytomegalovirus (HCMV) contains the cyclic AMP (cAMP) response element (CRE) consensus sequence, it was reasonable to hypothesize that cAMP might affect HCMV replication. Cyclic AMP modulating drugs such as 8-bromoadenosine 3',5'-cyclic monophosphate (BrA), and papaverine were used to affect the intracellular levels of cAMP, and the effects of the drugs on HCMV replication were studied. While papaverine effectively inhibited HCMV multiplication and DNA synthesis, BrA exerted little effect on the production of infectious HCMV yields. The synthesis of DNA in HCMV-infected cells appeared to be stimulated by BrA In order to understand the effect of cAMP on the expression of HCMV major IE gene, plasmid (pCMVIE/CAT) containing a reporter gene driven by HCMV IE promoter was transfected into either permissive human embryo lung (HEL) cells or nonpermissive cells. PL,Javerine, which has been reported to block the HCMV-induced increase in cAMP, reduced the expression of pCMVIE/CA T in permissive HEL cells. Treatment of transfected cells with BrA increased the expression of HCMV major IE promoter not only in HEL cells, but also in nonpermissive HeLa and Vero cells. Therefore, it seems that the expression of HCMV major IE gene is regulated by cAMP.