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1.
Materials (Basel) ; 17(5)2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38473670

ABSTRACT

A new heat source combination, consisting of a uniform body heat source and a tilted double ellipsoidal heat source, has been developed for cold metal transfer (CMT) wire-arc additive manufacturing of Mg-Gd-Y-Zn-Zr alloy. Simulations were conducted to analyze the temperature field and stress distribution during the process. The optimal combination of feeding speed and welding speed was found to be 8 m/min and 8 mm/s, respectively, resulting in the lowest thermal accumulation and residual stress. Z-axis residual stress was identified as the main component of residual stress. Electron Backscatter Diffraction (EBSD) testing showed weak texture strength, and Kernel Average Misorientation (KAM) analysis revealed that the 1st layer had the highest residual stress, while the 11th layer had higher residual stress than the 6th layer. Microhardness in the 1st, 11th, and 6th layers varies due to residual stress impacts on dislocation density. Higher residual stress increases dislocation density, raising microhardness in components. The experimental results were highly consistent with the simulated results.

2.
Materials (Basel) ; 15(24)2022 Dec 09.
Article in English | MEDLINE | ID: mdl-36556592

ABSTRACT

This paper investigated the effect of silicon phase morphology and size on microstructure, mechanical properties, and corrosion resistance of the AlSi10Mg alloys fabricated by selective laser melting (SLM). Using different heat treatment conditions for SLM-fabricated alloys, the microstructure characteristics and mechanical properties are analyzed. The corrosion behavior analysis is also performed using potentiodynamic polarization, electrochemical and immersion tests. Results show that the AlSi10Mg alloy directly fabricated by SLM has a continuous eutectic silicon network, which has a small driving force for corrosion and facilitates the deposition of corrosion products and generates a dense protective film. On the contrary, the formation of large isolated and uniformly distributed silicon particles produces a greater corrosion driving force after heat treatment, which makes most of the corrosion products transfer to the solution. The corrosion resistance of AlSi10Mg alloy directly fabricated by SLM is better than that of the alloys with heat treatment. Moreover, the heat treatment reduces the hardness of AlSi10Mg alloys due to the decrease in the solid solution strengthening effect.

3.
Materials (Basel) ; 15(4)2022 Feb 12.
Article in English | MEDLINE | ID: mdl-35207900

ABSTRACT

In order to obtain a high-performance heat-resistant Mg alloy during the rheo-rolling process, the electronic structure, elastic constants, binding energy and thermodynamic properties of the MgSnLa compounds were conducted by first-principle calculations. The results show that the MgSnLa compounds (La5Sn3, Mg17La2 and Mg2Sn) all show certain metallicity, and La5Sn3 has better mechanical properties (higher bulk modulus (46.47091 GPa) and shear modulus (26.40561 GPa)) than the other two phases. The binding energy reveals that La5Sn3 is the most stable phase in these composite phases (5.33 eV/atom); additionally, thermodynamic studies show that the structural stability of the MgSnLa compounds increases with the increase in temperature, and the temperature has the greatest effect on the stability of Mg17La2. These all provide an efficient guide for the widespread engineering applications of high-performance heat-resistant Mg alloy.

4.
Materials (Basel) ; 14(6)2021 Mar 23.
Article in English | MEDLINE | ID: mdl-33807033

ABSTRACT

TiB2/316L stainless steel composites were prepared by selective laser melting (SLM), and the adhesion work, interface energy and electronic structure of TiB2/γ-Fe interface in TiB2/316L stainless steel composites were investigated to explore the heterogeneous nucleation potential of γ-Fe grains on TiB2 particles using first principles. Six interface models composed of three different stacking positions and two different terminations were established. The B-terminated-top 2 site interface ("B-top 2") was the most stable because of the largest adhesion work, smallest interfacial distances, and smallest interfacial energy. The difference charge density and partial density of states indicated that a large number of strong Fe-B covalent bonds were formed near the "B-top 2" interface, which increased the stability of interface. Fracture analysis revealed that the bonding strength of the "B-top 2" interface was higher than that of the Fe matrix, and it was difficult to fracture at the interface. The interface energy at the Ti-poor position in the "B-top 2" interface model was smaller than that of the γ-Fe/Fe melt, indicating that TiB2 had strong heterogeneous nucleation potency for γ-Fe.

5.
Materials (Basel) ; 13(18)2020 Sep 08.
Article in English | MEDLINE | ID: mdl-32911692

ABSTRACT

AlSi10Mg alloy branches were fabricated by selective laser melting (SLM), and the branches were employed to evaluate their effect on the mechanical properties. When the porous branches were compressed along its building direction, the tree column structures-type AlSi10Mg alloy branches collapsed twice, which had typical elastic, shear, collapse, and densification stages. The compressive stress concentration at the interface between the support and the porous body caused the fracture of the tree column structures-type AlSi10Mg alloy branches. The fracture surface indicated that the prepared tree-type branches were distributed with different shapes of dimples, and the Si content inside the dimples was higher than that of the edge. The morphology of the Al-Si eutectic structure formed by SLM and the stress concentration at the Al/Al-Si-eutectic interface affected the fracture morphology and Si content distribution.

6.
Materials (Basel) ; 13(3)2020 Feb 04.
Article in English | MEDLINE | ID: mdl-32033202

ABSTRACT

The Al4C3 phase was precipitated via a reaction of graphene (Gr) with Al during selective laser melting (SLM). The interfacial nature of the Gr (0001)/Al4C3 (0001) interface was determined using the first-principle calculation. The simulation results showed that the influence of the stacking site on the interfacial structure was limited and the Al-termination interface presented a more stable structure than the C-termination interface. The Al-termination-CH site interface had the largest work of adhesion (6.28 J/m2) and the smallest interfacial distance (2.02 Å) among the four interfacial structures. Mulliken bond population analysis showed that the bonding of the Al-termination interface was a mixture of covalent and ionic bonds and there was no chemical bonding in the C-termination interface.

7.
Materials (Basel) ; 12(6)2019 Mar 21.
Article in English | MEDLINE | ID: mdl-30901913

ABSTRACT

In order to improve the abrasion performance of 316Lss, make full use of its advantages and broaden its application fields, the tribological behavior of the TiC particles reinforced 316Lss composites-which were manufactured by selective laser melting (SLM)-were investigated. In this study, GCr15 bearing steel was selected as the friction material and experiments on the sliding friction and wear under different loads of 15 N, 25 N and 35 N at the sliding speeds of 60, 80 and 100 mm/min were carried out, respectively. The results show that the wear performance of the TiC/316Lss composite is higher than that of the matrix during the friction and wear experiments under all conditions and the wear rate of the TiC/316Lss composite decreases with increasing the friction rate. Similar to the wear mechanism under different loads, it changes from abrasive wear to delamination wear and severe oxidative wear. At the same time, the mechanical mixed layer formed at a high speed has a protective effect on the matrix. The reason for this phenomenon is that the mechanical properties of the TiC/316Lss composites are significantly improved due to the addition of TiC particles, the refinement of cells near the TiC particles and the formation of a large number of dislocations. In addition, due to the presence of the TiC particles, the hardness and strength of the TiC/316Lss composites are greatly improved, thus the processing hardening ability of sub-surface has been improved.

8.
Materials (Basel) ; 12(2)2019 Jan 21.
Article in English | MEDLINE | ID: mdl-30669660

ABSTRACT

This research explored a novel chemical reduction of organic aluminum for plating Al on a graphene surface. The thermodynamics of the Al plating reaction process were studied. The Al plating process consisted of two stages: the first was to prepare (C2H5)3Al. In this reaction, the ΔH(enthalpy) was 10.64 kcal/mol, the ΔG(Gibbs free energy) was 19.87 kcal/mol and the ΔS(entropy) was 30.9 cal/(mol·K); this was an endothermic reaction. In the second stage, the (C2H5)3Al decomposed into Al atoms, which were gradually deposited on the surface of the graphene and the Al plating formed. At 298.15 K, the ΔH was -20.21 kcal/mol, the ΔG was -54.822 kcal/mol, the ΔS was 116.08 cal/(mol·K) and the enthalpy change was negative, thus indicating an endothermic reaction.

9.
Materials (Basel) ; 11(8)2018 Jul 30.
Article in English | MEDLINE | ID: mdl-30061504

ABSTRACT

In this research, the effect of several heat treatments on the microstructure and microhardness of TC4 (Ti6Al4V) titanium alloy processed by selective laser melting (SLM) is studied. The results showed that the original acicular martensite α'-phase in the TC4 alloy formed by SLM is converted into a lamellar mixture of α + ß for heat treatment temperatures below the critical temperature (T0 at approximately 893 °C). With the increase of heat treatment temperature, the size of the lamellar mixture structure inside of the TC4 part gradually grows. When the heat treatment temperature is above T0, because the cooling rate is relatively steep, the ß-phase recrystallization transforms into a compact secondary α-phase, and a basketweave structure can be found because the primary α-phase develop and connect or cross each other with different orientations. The residence time for TC4 SLM parts when the treatment temperature is below the critical temperature has little influence: both the α-phase and the ß-phase will tend to coarsen but hinder each other, thereby limiting grain growth. The microhardness gradually decreases with increasing temperature when the TC4 SLM part is treated below the critical temperature. Conversely, the microhardness increases significantly with increasing temperature when the TC4 SLM part is treated above the critical temperature.

10.
Materials (Basel) ; 11(9)2018 Aug 28.
Article in English | MEDLINE | ID: mdl-30154379

ABSTRACT

Ti/TiBCN composite coatings were prepared on a 7075 aluminum alloy surface by laser cladding. The relation between the main processing parameters (i.e., laser power, scanning speed, and powder feeding rate) and the geometrical characteristics (i.e., height, width, penetration depth, dilution and wetting angle) of single clad tracks is studied by linear regression analysis. The microstructure, micro-hardness and electrochemical corrosion were investigated by scanning electron microscopy, a Vickers micro-hardness machine, and a standard three-electrode cell, respectively. The results showed that all geometrical track characteristics are observed with high values of the correlation coefficient (R > 0.95). In addition, the average hardness value (750 HV0.2) was obtained of the Ti/TiBCN composite coating, and polarization curves indicated that the composite coatings were harder to corrode than the substrate.

11.
Materials (Basel) ; 11(9)2018 Aug 24.
Article in English | MEDLINE | ID: mdl-30149554

ABSTRACT

In this paper, GH4169 alloy's distributions of temperature and stress during the selective laser melting (SLM) process were studied. The SLM process is a dynamic process of rapid melting and solidification, and we found there were larger temperature gradients near the turning of scan direction and at the overlap of the scanning line, which produced thermal strain and stress concentration and gave rise to warping deformations. The stresses increased as the distance became further away from the melt pool. There was tensile stress in the most-forming zones, but compressive stress occurred near the melt pool area. When the parts were cooled to room temperature after the SLM process, tensile stress was concentrated around the parts' boundaries. Residual stress along the z direction caused the warping deformations, and although there was tensile stress in the parts' surfaces, but there was compressive stress near the substrate.

12.
Acta Biomater ; 48: 499-520, 2017 01 15.
Article in English | MEDLINE | ID: mdl-27746360

ABSTRACT

Crystalline Mg-Zinc (Zn)-Strontium (Sr) ternary alloys consist of elements naturally present in the human body and provide attractive mechanical and biodegradable properties for a variety of biomedical applications. The first objective of this study was to investigate the degradation and cytocompatibility of four Mg-4Zn-xSr alloys (x=0.15, 0.5, 1.0, 1.5wt%; designated as ZSr41A, B, C, and D respectively) in the direct culture with human umbilical vein endothelial cells (HUVEC) in vitro. The second objective was to investigate, for the first time, the early-stage inflammatory response in cultured HUVECs as indicated by the induction of vascular cellular adhesion molecule-1 (VCAM-1). The results showed that the 24-h in vitro degradation of the ZSr41 alloys containing a ß-phase with a Zn/Sr at% ratio ∼1.5 was significantly faster than the ZSr41 alloys with Zn/Sr at% ∼1. Additionally, the adhesion density of HUVECs in the direct culture but not in direct contact with the ZSr41 alloys for up to 24h was not adversely affected by the degradation of the alloys. Importantly, neither culture media supplemented with up to 27.6mM Mg2+ ions nor media intentionally adjusted up to alkaline pH 9 induced any detectable adverse effects on HUVEC responses. In contrast, the significantly higher, yet non-cytotoxic, Zn2+ ion concentration from the degradation of ZSr41D alloy was likely the cause for the initially higher VCAM-1 expression on cultured HUVECs. Lastly, analysis of the HUVEC-ZSr41 interface showed near-complete absence of cell adhesion directly on the sample surface, most likely caused by either a high local alkalinity, change in surface topography, and/or surface composition. The direct culture method used in this study was proposed as a valuable tool for studying the design aspects of Zn-containing Mg-based biomaterials in vitro, in order to engineer solutions to address current shortcomings of Mg alloys for vascular device applications. STATEMENT OF SIGNIFICANCE: Magnesium (Mg) alloys specifically designed for biodegradable implant applications have been the focus of biomedical research since the early 2000s. Physicochemical properties of Mg alloys make these metallic biomaterials excellent candidates for temporary biodegradable implants in orthopedic and cardiovascular applications. As Mg alloys continue to be investigated for biomedical applications, it is necessary to understand whether Mg-based materials or the alloying elements have the intrinsic ability to direct an immune response to improve implant integration while avoiding cell-biomaterial interactions leading to chronic inflammation and/or foreign body reactions. The present study utilized the direct culture method to investigate for the first time the in vitro transient inflammatory activation of endothelial cells induced by the degradation products of Zn-containing Mg alloys.


Subject(s)
Alloys/pharmacology , Human Umbilical Vein Endothelial Cells/pathology , Inflammation/pathology , Cell Adhesion/drug effects , Cell Death/drug effects , Corrosion , Culture Media , Electrochemical Techniques , Human Umbilical Vein Endothelial Cells/drug effects , Humans , Ions , Magnesium/pharmacology , Solubility , Spectrometry, X-Ray Emission , Surface Properties
13.
J Biomed Mater Res A ; 103(9): 2974-86, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25690931

ABSTRACT

Magnesium (Mg) alloy is an attractive class of metallic biomaterial for cardiovascular applications due to its biodegradability and mechanical properties. In this study, we investigated the degradation in blood, thrombogenicity, and cytocompatibility of Magnesium-Zinc-Strontium (Mg-Zn-Sr) alloys, specifically four Mg-4 wt % Zn-xSr (x = 0.15, 0.5, 1, and 1.5 wt %) alloys, together with pure Mg control and relevant reference materials for cardiovascular applications. Human whole blood and platelet rich plasma (PRP) were used as the incubation media to investigate the degradation behavior of the Mg-Zn-Sr alloys. The results showed that the PRP had a greater pH increase and greater concentration of Mg(2+) ions when compared with whole blood after 2 h of incubation with the same respective Mg alloys, suggesting that the Mg alloys degraded faster in PRP than in whole blood. The Mg alloy with 4 wt % Zn and 0.15 wt % Sr (named as ZSr41A) was identified as the most promising alloy for cardiovascular stent applications, because it showed slower degradation and less thrombogenicity, as indicated by the lower concentrations of Mg(2+) ions released and less deposition of platelets. Additionally, ZSr41 alloys were cytocompatible with fibroblasts in direct exposure culture in which the cells adhered and proliferated around the samples, with no statistical difference in cell adhesion density compared with the blank reference. Future studies on the ZSr41 alloys are necessary to investigate their direct interactions with other important cells in cardiovascular system, such as vascular endothelial cells and smooth muscle cells.


Subject(s)
Alloys/chemistry , Biocompatible Materials/chemistry , Absorbable Implants , Adult , Alloys/pharmacokinetics , Alloys/pharmacology , Animals , Biocompatible Materials/pharmacokinetics , Biocompatible Materials/pharmacology , Blood/drug effects , Blood Platelets/drug effects , Cells, Cultured , Fibroblasts/drug effects , Humans , In Vitro Techniques , Magnesium/chemistry , Materials Testing , Mice , Platelet Adhesiveness/drug effects , Platelet-Rich Plasma/drug effects , Strontium/chemistry , Surface Properties , Zinc/chemistry
14.
Acta Biomater ; 12: 298-321, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25449917

ABSTRACT

Crystalline Mg-Zn-Ca ternary alloys have recently attracted significant interest for biomedical implant applications due to their promising biocompatibility, bioactivity, biodegradability and mechanical properties. The objective of this study was to characterize as-cast Mg-xZn-0.5Ca (x=0.5, 1.0, 2.0, 4.0wt.%) alloys, and determine the adhesion and morphology of bone marrow derived mesenchymal stem cells (BMSCs) at the interface with the Mg-xZn-0.5Ca alloys. The direct culture method (i.e. seeding cells directly onto the surface of the sample) was established in this study to probe the highly dynamic cell-substrate interface and thus to elucidate the mechanisms of BMSC responses to dynamic alloy degradation. The results showed that the BMSC adhesion density on these alloys was similar to the cell-only positive control and the BMSC morphology appeared more anisotropic on the rapidly degrading alloy surfaces in comparison with the cell-only positive control. Importantly, neither culture media supplemented with up to 27.6mM Mg(2+) ions nor media intentionally adjusted up to alkaline pH 9 induced any detectable adverse effects on BMSC responses. We speculated that degradation-induced dynamic surface topography played an important role in modulating cell morphology at the interface. This study presents a clinically relevant in vitro model for screening bioresorbable alloys, and provides useful design guidelines for determining the degradation rate of implants made of Mg-Zn-Ca alloys.


Subject(s)
Alloys/pharmacology , Bone Marrow Cells/drug effects , Mesenchymal Stem Cells/drug effects , Alloys/chemistry , Animals , Calcium/chemistry , Calcium/pharmacology , Cells, Cultured , Female , Magnesium/chemistry , Magnesium/pharmacology , Microscopy, Electron, Scanning , Rats , Rats, Sprague-Dawley , Zinc/chemistry , Zinc/pharmacology
15.
Mater Sci Eng C Mater Biol Appl ; 33(7): 3661-9, 2013 Oct.
Article in English | MEDLINE | ID: mdl-23910262

ABSTRACT

A new biodegradable magnesium-zinc-strontium (Mg-Zn-Sr) alloy was developed and studied for medical implant applications. This first study investigated the alloy processing (casting, rolling, and heat treatment), microstructures, mechanical properties, and degradation properties in simulated body fluid (SBF). Aging treatment of the ZSr41 alloy at 175 °C for 8h improved the mechanical properties when compared to those of the as-cast alloy. Specifically, the aged ZSr41 alloy had an ultimate tensile strength of 270 MPa, Vickers hardness of 71.5 HV, and elongation at failure of 12.8%. The mechanical properties of the ZSr41 alloy were superior as compared with those of pure magnesium and met the requirements for load-bearing medical implants. Furthermore, the immersion of the ZSr41 alloy in SBF showed a degradation mode that progressed cyclically, alternating between pitting and localized corrosion. The steady-state average degradation rate of the aged ZSr41 alloy in SBF was 0.96 g/(m(2)·hr), while the pH of SBF immersion solution increased. The corrosion current density of the ZSr41 alloy in SBF solution was 0.41 mA/mm(2), which was much lower than 1.67 mA/mm(2) for pure Mg under the same conditions. In summary, compared to pure Mg, the mechanical properties of the new ZSr41 alloy improved while the degradation rate decreased due to the addition of Zn and Sr alloying elements and specific processing conditions. The superior mechanical properties and corrosion resistance of the new ZSr41 alloy make it a promising alloy for next-generation implant applications.


Subject(s)
Alloys/chemistry , Biomedical Technology , Materials Testing , Mechanical Phenomena , Alloys/pharmacology , Biodegradation, Environmental , Body Fluids/drug effects , Corrosion , Hardness/drug effects , Hydrogen-Ion Concentration/drug effects , Mechanical Phenomena/drug effects , Microscopy, Electron, Scanning , Spectrometry, X-Ray Emission , Surface Properties , Tensile Strength/drug effects , X-Ray Diffraction
16.
J Biomed Mater Res A ; 100(4): 999-1015, 2012 Apr.
Article in English | MEDLINE | ID: mdl-22307984

ABSTRACT

A novel biodegradable Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy was successfully produced using a series of metallurgical processes; including melting, casting, rolling, and heat treatment. The hardness and ultimate tensile strength of the alloy sheets increased to 71.2HV and 320 MPa after rolling and then aging for 12 h at 175°C. These mechanical properties were sufficient for load-bearing orthopedic implants. A hydroxyapatite (HA) coating was deposited on the Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy using a novel coating process combining alkali heat pretreatment, electrodeposition, and alkali heat posttreatment. The microstructure, composition, and phases of the Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy and HA coating were characterized using scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), and X-ray diffraction (XRD). The degradation, hemolysis, and cytocompatibility of the HA-coated and uncoated Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy were studied in vitro. The corrosion potential (E(corr)) of Mg-4.0Zn-1.0Ca-0.6Zr alloy (-1.72 V) was higher than Mg (-1.95 V), Mg-0.6Ca alloy (-1.91 V) and Mg-1.0Ca alloy (-1.97 V), indicating the Mg-Zn-Ca-Zr alloy would be more corrosion resistant. The initial corrosion potential of the HA-coated Mg alloy sample (-1.51 V) was higher than the uncoated sample (-1.72 V). The hemolysis rates of the HA-coated and uncoated Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy samples were both <5%, which met the requirements for implant materials. The HA-coated and uncoated Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy samples demonstrated the same cytotoxicity score as the negative control. The HA-coated samples showed a slightly greater relative growth rate (RGR%) of fibroblasts than the uncoated samples. Both the HA-coated and uncoated Mg-4.0Zn-1.0Ca-0.6Zr (wt %) alloy provided evidence of acceptable cytocompatibility for medical applications.


Subject(s)
Alloys , Cell Survival , Durapatite/chemistry , Electrochemical Techniques , Hemolysis , Calcium/chemistry , Humans , In Vitro Techniques , Magnesium/chemistry , Microscopy, Electron, Scanning , X-Ray Diffraction , Zinc/chemistry , Zirconium/chemistry
17.
Article in English | MEDLINE | ID: mdl-23366416

ABSTRACT

Magnesium-based alloys have attracted great interest for medical applications due to their unique biodegradable capability and desirable mechanical properties. When considered for medical applications, the degradation rate of these alloys must be tailored so that: (i) it does not exceed the rate at which the degradation products can be excreted from the body, and (ii) it is slow enough so that the load bearing properties of the implant are not jeopardized and do not conflict prior to and during synthesis of new tissue. Implant integration with surrounding cells and tissues and mechanical stability are critical aspects for clinical success. This study investigated Magnesium-Zinc-Strontium (ZSr41) alloy degradation rates and the interaction of the degradation products with human embryonic stem cells (hESC) over a 72 hour period. An in vitro hESC model was chosen due to the higher sensitivity of ESCs to known toxicants which allows to potentially detect toxicological effects of new biomaterials at an early stage. Four distinct ZSr41 compositions (0.15 wt.%, 0.5 wt.%, 1 wt.%, and 1.5 wt.% Sr) were designed and produced through metallurgical processing. ZSr41 alloy mechanical properties, degradation, and cytocompatibility were investigated and compared to pure polished Magnesium (Mg). Mechanical properties evaluated included hardness, ultimate tensile strength, and elongation to failure. Degradation was characterized by measuring total weight loss of samples and pH change in the cell culture media. Cytocompatibility was studied by comparing fluorescence and phase contrast images of hESCs after co-culture with Mg alloys. Results indicated that the Mg-Zn-Sr alloy with 0.15 wt.% Sr improved cytocompatibility and provided slower degradation as compared with pure Mg.


Subject(s)
Alloys/metabolism , Embryonic Stem Cells/metabolism , Humans , Hydrogen-Ion Concentration
18.
Protein Expr Purif ; 75(2): 133-7, 2011 Feb.
Article in English | MEDLINE | ID: mdl-20736068

ABSTRACT

Escherichia coli has proved to be a successful host for the expression of many heterologous proteins, and much efforts have been made toward improving recombinant protein expression including the usage of strong promoters and co-expression with chaperones. But little attention was paid on the relation between expression level and function of the target protein. Glycerophosphate oxidase (GPO) is a protein with FAD cofactor (without free cysteine and disulfide bonds).It was observed that the specific activity of GPO dramatically decreased with the increase of inducer IPTG. In addition, the stability of it decreased correspondingly. The structural difference of samples expressed under varying IPTG was investigated using size-exclusion and reverse-phase high performance liquid chromatography, together with CD spectrum. It was found that the conformation of peptide and organization of subunits were not affected. The loss of specific activity and stability were correlated to incomplete attachment of FAD onto GPO. These results revealed that synthesis speed should be controlled either by reduction of IPTG amount or using weak promoters in the production of GPO.


Subject(s)
Enterococcus faecium/enzymology , Escherichia coli , Glycerolphosphate Dehydrogenase , Isopropyl Thiogalactoside , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Chromatography , Circular Dichroism , Cloning, Molecular , Enterococcus faecium/genetics , Enzyme Stability , Flavin-Adenine Dinucleotide/metabolism , Gene Expression Regulation, Bacterial , Glycerolphosphate Dehydrogenase/genetics , Glycerolphosphate Dehydrogenase/metabolism , Holoenzymes/genetics , Holoenzymes/isolation & purification , Holoenzymes/metabolism , Isopropyl Thiogalactoside/metabolism , Protein Conformation , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Spectrum Analysis
19.
Transl Res ; 156(1): 1-6, 2010 Jul.
Article in English | MEDLINE | ID: mdl-20621031

ABSTRACT

Endothelial lipase, which is a newly identified member of the lipase family, plays an important role in high-density lipoprotein metabolism, which catalyzes the hydrolysis of high-density lipoprotein phospholipids and facilitates the clearance of high-density lipoprotein from the circulation. In addition, inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta), upregulate endothelial lipase expression, and endothelial lipase also affects the expression of cytokines, which in turn play an important role in atherogenesis. Endothelial lipase expression has been associated with macrophages within human atherosclerotic lesions. However, an important challenge is to determine how endothelial lipase alters the progression of atherosclerosis. Although few data are available from human studies, it seems that plasma endothelial lipase levels in individuals with atherosclerosis might be higher than that measured in healthy individuals. Therefore, we believe that endothelial lipase might be a promising marker for atherosclerosis in clinical settings in the future.


Subject(s)
Atherosclerosis/enzymology , Inflammation/enzymology , Lipase/blood , Lipoproteins/metabolism , Gene Expression Regulation, Enzymologic/physiology , Humans , Lipase/chemistry , Lipase/genetics
20.
Gen Comp Endocrinol ; 143(2): 184-92, 2005 Sep 01.
Article in English | MEDLINE | ID: mdl-15894316

ABSTRACT

Pituitary, a master gland of neuroendocrine system, secretes hormones that orchestrate many physiological processes, under the regulation of multiple signaling pathways. To investigate the genes involved in hormones expression of human pituitary, homemade cDNA microarray containing 14,800 human genes/ESTs were used to profile the gene expression in both fetal and adult pituitaries. Seven hundred and twelve known genes changed over 2-fold between the both tissues. Of which, 23 genes were changed with hormones expression in aging were confirmed by RT-PCR, not only the known regulators such as Pit1, GATA4, ESRRA, GABA-A, and EMK, but also LOC55884, DUSP3, PNN, and RCL, which had not been reported to be involved in the hormones expression. Correspondingly, the mRNAs of GH, PRL, POMC, TSH-beta, FSH-beta, and LH-beta, was increased as much as 6- to 20-fold in adult pituitary than those in fetal pituitary, by real-time quantitative RT-PCR assay. In addition, the mRNAs of signaling pathways, such as cAMP-PKA-CREB, PI3K-Akt, and PKA-ERK were further investigated. Of them, it was only cAMP-PKA-CREB pathway, but not PI3K-Akt and PKA-ERK have the same expressing pattern as hormones. It suggested that cDNA microarray is highly advantages to profile the differential expressed genes that were involved in hormones expression of human pituitary, but it might ignore some responding proteins regulated posttranscriptionally.


Subject(s)
Gene Expression Profiling , Gene Expression Regulation, Developmental/genetics , Oligonucleotide Array Sequence Analysis , Pituitary Gland/metabolism , Pituitary Hormones/genetics , Signal Transduction/genetics , Adult , Aged , Aging/genetics , Aging/physiology , Female , Fetus/metabolism , Humans , Male , Middle Aged , Pituitary Hormones/metabolism , RNA, Messenger/analysis , Signal Transduction/physiology
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