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ABSTRACT: Decitabine is a synthesized cytosine analog that is a potent inhibitor of DNA methylation. There have been a few reports on the in vitro anti-melanoma effect of decitabine or its functional mechanisms. We investigated the anti-proliferation effect of decitabine on the cultured murine melanoma cell line K1735M2. MTT assay showed that decitabine had strong inhibition on melanoma K1735M2 in a time- and dose-dependent manner in vitro. Morphological observation showed that decitabine could induce melanoma K1735M2 cells to produce dendrite-like structures with the increase of decitabine concentration and incubation time. Decitabine could effectively induce K1735M2 cells to differentiate in vitro. Additionally, decitabine could induce a dose-dependent G2/M cell cycle arrest in K1735M2 cells. We provided experimental evidences that the anti-proliferation effect of decitabine on murine K1735M2 melanoma cells was associated predominately with G2/M cell cycle arrest and the induction of differentiation rather than apopotosis.
Environmental toxicology and pharmacology. 11/2011; 32(3):423-9.
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ABSTRACT: The nacreous layer of molluskan shells, which consists of highly oriented aragonitic crystals and an organic matrix (including chitin and proteins), is a product of biomineralization. This paper briefly introduces the recent research advances on nacre biomineralization of shells from bivalves and gastropods, which mainly focus on analysis of the micro- and nano-structure and components of shell nacreous layers, and investigations of the characteristics and functions of matrix proteins from nacre. Matrix proteins not only participate in construction of the organic nacre framework, but also control the nucleation and growth of aragonitic crystals, as well as determine the polymorph specificity of calcium carbonate in nacre. Moreover, the inorganic aragonite phase also plays an active role in organizing nacre microstructure. Based on these studies, several models to illustrate the formation mechanism related to lamellar nacre in bivalves, and columnar nacre in gastropods are introduced.
Progress in molecular and subcellular biology 01/2011; 52:331-52.
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ABSTRACT: To examine the in vivo effects of atorvastatin (AT) on arterial calcification in rats, arterial calcification was established by subcutaneous injection of vitamin D3 and Warfarin. Intragastric administration of AT began 4 days before establishment of arterial calcification in the AT group (n=6). Blood samples were taken and abdominal aortas were collected and stained. After induction of calcification, plasma Ca(2+) levels in the CA and AT groups were significantly higher than those before treatment and in the untreated controls. Plasma Ca(2+) levels in the AT group were significantly lower than in the CA group. The relative calcification area in aortic specimens from the AT group was significantly smaller than in the CA group. Rat aortic vascular smooth muscle cells (VMSC) were isolated from abdominal aortic segments and pre-treated with AT (1, 5, or 10 μM) for 24 hr. Cells in the calcification (CA) group and the AT group were cultured with β-glycerophosphate, insulin and vitamin C for 14 days to induce cell calcification. Calcium deposition and alkaline phosphatase activity were significantly increased in the CA group compared to untreated controls (p<0.01). This effect was ameliorated by AT (all p<0.01). In vivo administration of AT reduced arterial calcification and plasma Ca(2+) concentration. In vitro, AT reduced calcification markers in rat aortic vascular smooth muscle cells.
Basic & Clinical Pharmacology & Toxicology 10/2010; 107(4):798-802. · 2.18 Impact Factor
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ABSTRACT: Our study investigated a rapid and reliable method of surgical occlusion of coronary vessels in mice. The study improves the chance of success in making a myocardial ischemia model in mice, and provides a novel method to a novice and laboratory with limited conditions.
Sixty mice were evenly divided into two groups, a modified group (oral intubation and self-made rib retractor) and a conventional group (tracheotomy and rib cutting). During the perioperative period, the success rate of model establishment and the survival rate of the mice in the modified group were significantly higher than those in the conventional group (P<0.01). Also, the status of the mice in the modified group after operation was better than that of the conventional group. Moreover, operation times in the modified group were significantly shorter than those of the conventional group (P< 0.01). The infarct size, as assessed using triphenyltetrazolium chloride staining, was similar between the two groups (P> 0.05).
This novel method is simple and efficient and can be conducted independently, enhancing the success rate of myocardial ischemic model establishment in mice.
Journal of Medical Systems 06/2010; 34(3):413-7. · 1.13 Impact Factor
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ABSTRACT: We investigated possible mechanism(s) where honokiol induces apoptosis in human hepatocellular carcinoma SMMC-7721 cells. MTT assay showed that honokiol has strong inhibition on SMMC-7721 cells in a dose-dependent manner. SMMC-7721 cells after honokiol treatment display morphological characteristics such as cell shrinkage, detachment from the culture plate, formation of apoptotic bodies, change to a round shape, and marked nuclear condensation and fragmentation after 32258 staining. Cell apoptosis was measured by Annexin-V/PI staining and alternatively, by the subG0/G1 percentage of the cell cycle analysis followed by FACS. An obvious loss of ΔΨ(m) and a quick burst of ROS was detected when honokiol reached 4μg/ml, which was coincident with the high apoptosis percentage in our previous research. Up-regulation of Bax and down-regulation of Bcl-2 were observed, suggesting that honokiol-induced apoptosis was associated with reactive oxygen species (ROS) production and an increase of Bax/Bcl-2 ratios.
Environmental Toxicology and Pharmacology 07/2009; 28(1):97-103. · 1.47 Impact Factor
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ABSTRACT: To shorten operation time and improve survival rate of rats with myocardial ischemia or myocardial infarction, we use a novel device comprised of a face mask and a head/neck retainer in this study. We report the basic design of the novel respiratory face mask (RFM) and evaluate its performance in a rat model of myocardial ischemia. The device is cost-effective and easier to handle than other devices, such as tracheal intubation. Compared with conventional tracheal intubation, we found that RFM shortens operation time significantly while keeping blood indices normal; the mean operation time for rats in the mask group was (32+/-3) min, and that for the intubation group was (45+/-7) min (P<0.05). Moreover, the size and shape of the RFM can be changed according to the body weight of rats. In conclusion, RFM is an appropriate device for the establishment of myocardial infarction or ischemia-reperfusion in rats.
Journal of Zhejiang University SCIENCE B 05/2009; 10(5):391-4. · 1.10 Impact Factor
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ABSTRACT: To investigate the antagonism of LY333531 on the increased permeability of cardiac microvascular endothelial cells (CMECs) induced by high glucose.
The cultured CMECs from rats were randomly divided into four groups: normal group, high glucose group (25 mmol/L), high glucose+LY333531 (10 micromol/L) group and high glucose+saline group. The permeability of cell monolayer was detected using in vitro vascular permeability assay kit. Cell apoptosis was determined by TUNEL and the expression of PKCbeta II was analyzed by immunofluorescence and Western blot in each group.
Compared with normal group, the permeability (400.0+/-20.00 vs 223.3+/-25.17; P<0.01) of cell monolayer cultured in high glucose medium was increased at a higher apoptosis rate (55.00%+/-5.000% vs 2.333%+/-1.155%; P<0.01) and PKCbeta II expression (0.4767+/-0.0751 vs 0.1733+/-0.0208; P<0.01). However, the high glucose+LY333531 group showed noticeable attenuation on both permeability (360+/-17.32 vs 400.0+/-20.00; P<0.05) and apoptosis (25.00%+/-5.000% vs 55.00%+/-5.000%; P<0.01) with reduced PKCbeta II expression (0.2800+/-0.0700 vs 0.4767+/-0.0751; P<0.01). No significant effects of saline on the cell permeability, apoptosis and PKCbeta II expression were observed.
The antagonism of LY333531 has shown obvious effects on the impairment of high glucose to the permeability of CMECs.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology 04/2009; 25(4):303-5.
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ABSTRACT: The present study defines the expression of Toll-like Receptor 2 (TLR2), and the modulatory role of Glycogen synthase kinase (GSK)-3beta inhibitor on TLR2/Nuclear Factor-kappa B (NF-kappaB) signaling following myocardial ischemia-reperfusion (MI-R) injury in rats.
Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) were used to analyze the presence and quantity of TLR2 mRNA and protein. Tumor necrosis factor (TNF)-alpha mRNA and interleukin-6 (IL-6) mRNA were analyzed by RT-PCR. The activation of NF-kappaB was detected by Western Blot and the myocardial infarct size by Evans blue-TTC staining.
Following 30 min of myocardial ischemia, a significant up-regulation of TLR2 mRNA was revealed by RT-PCR from 1 to 24 h post reperfusion. IHC demonstrated high protein expression levels of TLR2. Administration of the GSK-3beta inhibitor 4-benzyl-2-methyl-1, 2, 4-thiadiazolidine-3, 5-dione (TDZD-8) 5 min prior to reperfusion following 1 h reperfusion down-regulated mRNA levels of TLR2 and downstream proinflammatory cytokines (P < 0.05 vs. MI-R), decreased the activity of NF-kappaB and the size of the myocardial infarct (P < 0.05 vs. MI-R).
Our results demonstrate that TLR2 and its signaling components are activated by MI-R. TDZD-8 administration attenuates TLR2/NF-kappaB signaling, suggesting a possible mechanism whereby GSK-3beta inhibition improves the outcome of MI-R.
Agents and Actions 03/2009; 58(7):377-83. · 1.59 Impact Factor
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ABSTRACT: To investigate the effects of constant magnetic field (CMF) on proliferation and migration of bone marrow-derived endothelial progenitor cells (EPCs) under rapamycin intervention.
EPCs were isolated from rat bone marrow by density gradient centrifugation and cultured on fibronectin-coated dishes. Six days later the attached cells were divided into 5 groups: control group, rapamycin (1 ng/ml) group, and 3 rapamycin + CMF groups (treated with CMF of the doses 0.1 mT, 0.5 mT, and 1.0 mT respectively). Samples were collected 24 hours after incubation. Cell proliferation was measured by MTT chromatometre. EPC migration was detected with modified Boyden chamber assay.
The EPC proliferation ability of the rapamycin group, expressed by absorbance, was (0.252 +/- 0.006), significantly lower than that of the control group [(0.328 +/- 0.025), P < 0.05]. The number of migrating EPC was (31 +/- 3) cells, significantly lower than that of the control group [(48 +/- 5), P < 0.05]. The EPC proliferation ability of the rapamycin + CMF 0.5 mT and 1.0 mT groups, expressed by absorbance, were (0.278 +/- 0.008) and (0.280 +/- 0.010) respectively, both significantly higher than that of the control group (both P < 0.05). The migrating EPC number of the rapamycin + CMF 0.5 mT and 1.0 mT groups were (37 +/- 3) and (38 +/- 4) respectively, both significantly higher than that of the control group (both P < 0.05).
CMF of the doses of 0.5 mT and 1.0 mT antagonizes the effects of rapamycin on EPCs, increasing the proliferation and migration of EPCs.
Zhonghua yi xue za zhi 10/2008; 88(38):2719-21.
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ABSTRACT: SO3 belongs to the O-superfamily of conotoxins and is known to have analgesic effects in experimental animals. In order to explore the mechanism of its potential pharmacological actions, the effect of SO3 on synchronized spontaneous calcium spikes was examined in cultured hippocampal networks by calcium imaging. Spontaneous oscillations of intracellular concentrations of calcium (Ca(2+)) in the form of waves and spikes are found in cultured hippocampal networks. Exposure to increasing concentrations of SO3 resulted in a progressive decrease in synchronized spontaneous calcium spikes. The higher concentrations (0.1 micromol/L and 1 micromol/L) of SO3 showed the strongest inhibition. The rank order of inhibition was 1 micromol/L > 0.1 micromol/L > 10 micromol/L > 0.01 micromol/L. This action of SO3 in reducing synchronized calcium spikes suggests a possible application for therapeutic treatment of epilepsy.
Cell Biology and Toxicology 02/2008; 24(1):11-7. · 2.51 Impact Factor
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ABSTRACT: Alkaline phosphatases are ubiquitous enzymes found in most species including the pearl oyster, Pinctada fucata, where it is presumably involved in nacreous biomineralization processes. In the present study, we have purified alkaline phosphatases from the pearl oyster and modified the tryptophan residues using N-bromosuccinimide (NBS). We show that the resulting inactivation of purified alkaline phosphatase by NBS is dependent on modification of only one of five tryptophan residues in the enzyme. Substrate protection experiments showed that the tryptophan residue was not located at the substrate-binding site but was involved in the catalytic activity.
Biochemistry (Moscow) 02/2008; 73(1):87-91. · 1.06 Impact Factor
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ABSTRACT: A spherical symmetric design-response surface methodology was applied to optimize the preparation of daidzein-loaded chitosan microspheres by the emulsification/chemical cross-linking technique. The influence of polymer concentration, ratio of drug to polymer, and the stirring speed on the encapsulation efficiency, particle size, particle size distribution, and accumulative drug release percent in microspheres were evaluated. Scan electron microscopy of the optimized microspheres showed spherical particles, loading with drug microcrystal uniformly on the surface of and inside the microspheres. In vivo pharmacokinetic characteristics were evaluated after intramuscular injection of the microspheres in rats. The time-resolved fluoroimmunoassay method was used to determine plasma concentrations of daidzein. The data showed that the release of daidzein in the microspheres in vitro and in vivo almost lasted for 35 days. The bioavailability of daidzein in the microspheres by intramuscular injection increased up to 39% in rats, suggesting that the cross-linked chitosan microspheres are a valuable system for the long-term delivery of isoflavones.
International Journal of Pharmaceutics 07/2007; 338(1-2):142-51. · 3.35 Impact Factor
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ABSTRACT: To elucidate the mechanism of nacre biomineralization, the mantle of Pinctada fucata (P. fucata) from the South China Sea was used. Using the mantle cDNA library and the ESTs we have cloned through suppression subtractive hybridization (SSH), ten novel genes including PFMG1 were obtained through nested PCR. Bioinformative results showed that PFMG1 had a high homology (40%) with Onchocerca volvulus calcium-binding protein CBP-1 and had two EF-hand calcium-binding domains from the 81st to the 93rd amino acid and from the 98th to the 133rd amino acid in the deduced amino acid sequence. The results of multitissue RT-PCR and in situ hybridization demonstrated the high expression of PFMG1 in the mantle of P. fucata and confirmed the SSH method. The results of GST-PFMG1 on CaCO3 crystallization showed significant effects on nucleation and precipitation of CaCO3. PFMG1 was cloned into the pcDNA.3.1/myc-HisA vector and was subsequently transfected into MC3T3-E1 cells. RT-PCR revealed upregulation of the marker genes related to cell growth, differentiation, and mineralization, and BMP-2, osterix, and osteopontin were upregulated as a result. This research work suggests that PFMG1 plays an important role in the nacre biomineralization, and the SSH method can pave the way for the bulk cloning and characterization of new genes involved in biomineralization in P. fucata and may accelerate research on the mechanism of pearl formation.
Biochemistry 02/2007; 46(3):844-51. · 3.42 Impact Factor
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ABSTRACT: To prepare an inclusion complex of daidzein and hydropropyl-beta-cyclodextrin to enhance the solubility of daidzein.
The inclusion complex of daidzein was prepared by the solution stirring method. The binary system of daidzein and HP-beta-CD was confirmed by differential thermal, thermogravimetry analysis, infrared spectroscopy and X-ray diffractometry.
The drug content in the inclusion complex was 6. 76% and the solubility was 13.68 mg x mL(-1). The identification results showed that the inclusion complex was formed.
The preparation method of the inclusion complex of daidzein and hydropropyl-beta-cyclodextrin is simple and available, with a increased solubility of daidzein.
Zhongguo Zhong yao za zhi = Zhongguo zhongyao zazhi = China journal of Chinese materia medica 01/2007; 31(24):2039-41.
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ABSTRACT: To study the effect of alendronate on artery calcification in rats.
(1) 4-week SD male rats were randomly divided into 3 groups: alendronate group (AL, n = 6), calcification group (CA, n = 6) and normal group (N, n = 6). In AL and CA group, artery calcification of rat was established by subcutaneous injection of vitamin D3 (300,000 U x kg(-1) x d(-1) for 3 days) and Warfarin (15 mg x 100 g(-1) x 12 h(-1) for 4 days); In AL group, at 4 days before establishment of artery calcification, alendronate (1 mg x kg(-1) x 24 h(-1)) was administered with subcutaneous injection and continued to be given to the end of the study. Abdominal aortae were collected for paraffin section and stained with von Kossa staining to observe the area of calcification. (2) Rat aortic vascular smooth muscle cells (VSMC) were cultured in vitro with tissue explant. All cells were divided into 5 groups: normal group, calcification group (control group), and alendronate 10(-9), 10(-7) and 10(-5) mol/L group. Before inducing calcification, alendronate 10(-9), 10(-7) and 10(-5) mol/L group were individually pre-treated with final concentrations of 10(-9), 10(-7) and 10(-5) mol/L alendronate for 24 hours. Beta-glycerophosphate were then added in the calcification group and in all the alendronate groups to induce VSMC calcification. All cells were cultured for 14 days. Cell crawling slice was applied to Alizarin red S staining to observe VSMC calcification. Colorimetric method was applied to measure the contents of Ca2+, cell proteins, and ALP activity. The ratio of contents of Ca2+ and cell proteins was cell calcium deposits. Cell proliferation was measured with tetrazolium salt (MTT) method.
(1) With von Kossa staining the black deeply stained structure was found to be decreased in AL group. (2) As compared with the control group, in all the alendronate groups, showed that the number of calcium nodules [(6.8 +/- 2.7, 6.2 +/- 4.2, 5.3 +/- 2.4) % vs (7.4 +/- 3.8)%], and cell calcium depositions [(5.2 +/- 1.2, 4.8 +/- 1.7, 3.5 +/- 1.8)% vs (5.6 +/- 1.6)%], cell ALP activity and cell proliferation decreased significantly and dose-dependently.
Alendronate can inhibit the artery calcification in rats.
Zhonghua nei ke za zhi [Chinese journal of internal medicine] 07/2006; 45(6):489-92.
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ABSTRACT: To evaluate the possibility and efficiency of nanoparticle as a new vector in vascular endothelial growth factor (VEGF) gene transference, and investigate the efficacy of direct gene transfer of nanoparticle with VEGF(165) gene into ischemic myocardium.
Nanoparticle-VEGF (Np/VEGF) complex was prepared with poly (D, L-lactide-co-glycolide) (PLGA) loading VEGF(165) gene and the envelopment efficiency and size of the complex were determined. The Np/VEGF was transfected into the cultured myocardial cells, RT-PCR and ELISA were used to evaluate the transfection of VEGF. Suspension of Np/VEGF was injected into the myocardial tissue of 4 rabbits. 96 hours after operation myocardial tissue was obtained, made into sections, and observed with electron microscope. New Zealand White rabbits underwent thoracotomy followed by ligation of left anterior descending coronary artery to establish ischemic models. The New Zealand White rabbits were divided into 3 groups: Np/VEGF group (n = 12, nanoparticle with VEGF(165) were injected into the cordial myocardium), blank plasmid group (n = 12, injected with blank VEGF(165) plasmid), and control group (n = 8, injected with normal saline). Ultrasonography and immunohistochemistry with factor VIII related antigen were conducted to evaluate the cardiac function and the collateral circulation of the occluded artery. One month later the rabbits were killed to observe the vascularization of capillaries in the ischemic myocardium.
The envelopment efficiency of the Np/VEGF complex thus prepared, 50 - 300 nm in size, were 1.87% y. RT-PCR and ELISA showed that VEGF gene had been successfully transfected into myocardial cells by the nanoparticles. A great number of nanoparticles were observed in the myocardial cytoplasm and nuclei. One month after operation, the ventricular wall motor amplitude of the Np/VEGF group was 1.87 mm +/- 0.32 mm, significantly larger than those of the blank plasmid group (1.59 mm +/- 0.24 mm, P < 0.05) and control group (0.93 mm +/- 0.40 mm, P < 0.05); and the left ventricular ejection fraction of the Np/VEGF group was 60% +/- 10%, significantly higher than those of the blank plasmid group (50% +/- 6%, P < 0.05) and control group (40% +/- 8%, P < 0.05). The capillary density at low power field (x 100) of the Np/VEGF group was 57 +/- 12, significantly higher than those of the VEGF group (41 +/- 14) and control group (24 +/- 8).
Nanoparticle can act as a vector to transfect specific gene in vitro and in vivo. Direct gene transfer of nanoparticle with DNA encoding VEGF into the ischemic rabbit myocardium can increase capillary number; therefore it may be a novel therapeutic approach for myocardial ischemia.
Zhonghua yi xue za zhi 03/2006; 86(8):510-4.
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ABSTRACT: In this study, we report that the steroid extract 5alpha, 8alpha-epidioxycholest-6-ene-3beta-ol (MME) from Meretrix meretrix has the ability to inhibit growth of hepatoma cells and to induce G1-phase cell cycle arrest in two human hepatoma cell lines, HepG2 and Hep3B. HepG2 cells were more sensitive than Hep3B to MME. The extract markedly up-regulated the expression of p53 and p21WAF1/CIP1 in HepG2, suggesting that MME-induced G1 phase cell cycle arrest in HepG2 might be p53-dependent. Therefore, the up-regulation of p27KIP1and p16INK4A in both cell lines indicates that a p53-independent pathway might be involved in the mechanism of MME inducing cell cycle arrest. In conclusion, MME induces G1 phase cell cycle arrest via both p53-dependent and p53-independent pathways.
Cancer Letters 03/2006; 232(2):199-205. · 4.24 Impact Factor
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ABSTRACT: Oridonin, a diterpenoid isolated from Rabdosia rubescens (Hemsl.) Hara, is currently one of the most important traditional Chinese herbal medicines. We investigated the anti-proliferation effect of oridonin on the cultured murine melanoma cell line K1735M2. The growth inhibition activity of oridonin for K1735M2 cells occurred in a time- and dose-dependent manner (IC50 was 7.4+/-0.6 microM). Further studies showed that these inhibition effects were associated with dose-dependent G2/M phase arrest and differentiation induction. Detection of morphological observation showed that oridonin could induce K1735M2 cells to produce dendrite-like structures. The results of the migration indicated that oridonin affected motility of K1735M2 cells in a dose-dependent manner. These results suggested that oridonin is a potential candidate for melanoma cancer therapy.
Journal of Ethnopharmacology 02/2006; 103(2):176-80. · 3.01 Impact Factor
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ABSTRACT: Alkaline phosphatases (ALP, EC 3.1.3.1) are ubiquitous enzymes found in most species. ALP from a pearl oyster, Pinctada fucata (PALP), is presumably involved in nacreous biomineralization processes. Here, chemical modification was used to investigate the involvement of basic residues in the catalytic activity of PALP. The Tsou's plot analysis indicated that the inactivation of PALP by 2,4,6-trinitrobenzenesulfonic acid (TNBS) and phenylglyoxal (PG) is dependent upon modification of one essential lysine and one essential arginine residue, respectively. Substrate reaction course analysis showed that the TNBS and PG inactivation of PALP followed pseudo-first-order kinetics and the second-order inactivation constants for the enzyme with or without substrate binding were determined. It was found that binding substrate slowed the PG inactivation whereas had little effect on TNBS inactivation. Protection experiments showed that substrates and competitive inhibitors provided significant protection against PG inactivation, and the modified enzyme lost its ability to bind the specific affinity column. However, the TNBS-induced inactivation could not be prevented in presence of substrates or competitive inhibitors, and the modified enzyme retained the ability to bind the affinity column. In a conclusion, an arginine residue involved in substrate binding and a lysine residue involved in catalysis were present at the active site of PALP. This study will facilitate to illustrate the role ALP plays in pearl formation and the mechanism involved.
The International Journal of Biochemistry & Cell Biology 08/2005; 37(7):1446-57. · 4.63 Impact Factor
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ABSTRACT: A new biphenyl derivative, 4',5-dimethoxy-3-beta-D-glucopyranosyloxy-4-hydroxy-biphenyl, named kakispyrol (1), has been isolated from the leaves of Diospyros kaki, together with three known compounds, vitexin (2), 2'-O-rhamnosyl vitexin (3) and isorhamnetin-3-O-beta-D-glucopyranoside (4). The structure of compound 1 has been determined on the basis of spectroscopic evidence.
Journal of Asian Natural Products Research 07/2005; 7(3):265-8. · 0.94 Impact Factor
