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ELISA Kit for Matrix Metalloproteinase 3 (MMP3) Organism: Equus caballus; Equine (Horse)

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[#E90101Eq] ELISA Kit for Matrix Metalloproteinase 3 (MMP3) Organism: Equus caballus; Equine (Horse)


E90101Eq | ELISA Kit for Matrix Metalloproteinase 3 (MMP3) Organism: Equus caballus; Equine (Horse), 96T
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(1) Controllable and durable release of BMP-2-loaded 3D porous sulfonated polyetheretherketone (PEEK) for osteogenic activity enhancement.[TOP]

Pubmed ID :30107340
Publication Date : //
Polyetheretherketone (PEEK) is ideal for dental and orthopedic applications because its mechanical properties are similar to cortical bones. However, its inherent inert ability hinders its clinical applications. In this work, bone morphogenetic protein-2 (BMP-2) was immobilized onto the sulfonated PEEK (SPEEK) using lyophilization technology. The surface morphologies of the samples were analyzed by field-emission scanning electron microscopy (FE-SEM), and the chemical compositions were analyzed by energy-dispersive X-ray spectrometry (EDS). The release content of BMP-2 of the samples immersed in the PBS (pH = 7.4) was detected by a human BMP-2 ELISA kit. The results indicated that controllable and durable BMP-2 release was accomplished due to the three-dimensional (3D) network of sulfonated PEEK. The in vitro cellular experiments showed that the BMP-2-immobilized samples significantly enhanced the initial adhesion and spreading of rat bone mesenchymal stem cells (rBMSCs). Moreover, the collagen secretion, extracellular matrix mineralization and ALP activity were also improved. Thus, the BMP-2-immobilized samples greatly promoted the osteogenic differentiation of rBMSCs, which revealed that BMP-2 immobilization paves the way for the use of PEEK in clinical applications.

Authors : Sun Zhenjie, Ouyang Liping, Ma Xiaohan, Qiao Yuqin, Liu Xuanyong,

(2) Exhaled matrix metalloproteinase 9 in patients with stable COPD - an observational study.[TOP]

Pubmed ID :30057384
Publication Date : //
Introduction Data on the measurement of matrix metalloproteinase 9 (MMP-9) in exhaled breath condensate (EBC) from patients with COPD are scarce and inconsistent. Objectives to 1/ assess the feasibility and agreement between enzyme-linked immunosorbent assay (ELISA) and immunoenzymatic assay (IEA) in MMP-9 measurement in EBC, and 2/ assess the relationships between EBC total and active MMP-9 and clinical and functional COPD characteristics. Patients and methods Total (ELISA and IEA) and active (IEA) MMP-9 were assessed in EBC from 70 patients with stable COPD and 21 controls and correlated with pulmonary function and COPD symptom level. Results EBC MMP-9 level did not reach the sensitivity threshold of the applied ELISA kit in all COPD patients and 11 controls.  EBC total and active MMP-9 (IEA) did not differ between COPD patients and controls. In COPD patients, total MMP-9 correlated positively with FEV1 and FEV1/FVC and inversely with RV/TLC. A weak positive correlation between EBC active MMP-9 concentration and CAT score (r = 0.31, p = 0.01) was found. Conclusions the utility of ELISA in the assessment of MMP-9 in EBC  is limited in patients with COPD. The measurement of EBC MMP-9 by IEA is feasible in these patients. The positive correlation between EBC active MMP-9 form and CAT score in our patients, as well as the inverse relationship between EBC total MMP-9 concentration and the degree of airway obstruction reflect the complex role of MMP-9 in COPD. Our results also point to some technical considerations that need to be taken into account in MMP-9 evaluation in EBC from patients with this disease.

Authors : Maskey-Warzęchowska Marta, Górska Katarzyna, Nejman-Gryz Patrycja, Paplińska-Goryca Magdalena, Grzela Tomasz, Krejner Alicja, Grzela Katarzyna, Krenke Rafał,

(3) TIMP‑3 suppresses the proliferation and migration of SMCs from the aortic neck of atherosclerotic AAA in rabbits, via decreased MMP‑2 and MMP‑9 activity, and reduced TNF‑α expression.[TOP]

Pubmed ID :29956789
Publication Date : //
The present study investigated the role of tissue inhibitor of matrix metalloproteinase‑3 (TIMP‑3) in regulating the proliferation, migration, apoptosis and activity of matrix metalloproteinase (MMP)‑2 and ‑9, during the development of an atherosclerotic abdominal artery aneurysm (AAA). Experiments were conducted using rabbit AAA neck (NA) smooth muscle cells (SMCs), to investigate the potential for TIMP‑3 to be used as a novel stent coating in preventing aortic dilation adjacent to the AAA. The atherosclerotic AAA model was induced in New Zealand white rabbits via a 6‑week high‑cholesterol diet, followed by incubation of the targeted aortic region with elastase. SMCs were isolated from the aorta adjacent to the aneurysm 30 days after AAA model induction, and stimulated with 3, 10, 30 or 100 ng/ml TIMP‑3. Cell proliferation was investigated using Cell Counting Kit‑8 reagent, migration was examined using a Boyden chamber assay and apoptotic rate was analyzed using the Annexin V‑fluorescein isothiocyanate Apoptosis Detection kit. Gelatin zymography and ELISA were used to measure the activity of MMP‑2 and MMP‑9, and the expression of tumor necrosis factor‑α (TNF‑α), respectively. Analysis of cell proliferation indicated that 10, 30 and 100 ng/ml TIMP‑3 reduced cell viability. Cell migration was decreased by 10, 30 and 100 ng/ml TIMP‑3. MMP‑2 activity was inhibited by 10, 30 and 100 ng/ml TIMP‑3, and MMP‑9 activity was suppressed by 30 and 100 ng/ml TIMP‑3. The protein levels of secreted TNF‑α were reduced by 10, 30 and 100 ng/ml TIMP‑3. The present study demonstrated the ability of 30 and 100 ng/ml TIMP‑3 to attenuate migration and proliferation, and to inhibit the activity of MMP‑2, MMP‑9 and TNF‑α secretion of NA SMCs. In conclusion, TIMP‑3 may be considered a potential therapeutic drug for use in a novel drug‑eluting stent, to attenuate the progressive dilation of the aortic NA.

Authors : Zhai Huan, Qi Xun, Li Zixuan, Zhang Wei, Li Chenguang, Ji Lu, Xu Ke, Zhong Hongshan,

(4) Uveal melanocytes express high constitutive levels of MMP-8 which can be upregulated by TNF-α via the MAPK pathway.[TOP]

Pubmed ID :29935949
Publication Date : //
Matrix metalloproteinase (MMP)-8 is the most potent MMP for degrading collagen type-1 and plays an important role in inflammatory reactions and tissue remolding processes. MMP-8 is expressed mainly by polymorphonuclear leukocytes and is not expressed constitutively by most non-leukocytes. We studied the constitutive and TNF-α-induced expression of MMP-8 in cultured human uveal melanocytes (UM) and the relevant signal pathways involved. Conditioned media and cells were collected from UM and other cell types. MMP-8 proteins and mRNA were measured using ELISA kit, western blot and real time RT-PCR, respectively. Phosphorylated p38 MAPK, ERK1/2, and JNK1/2 were measured by ELISA kit and western blot. Very high levels of MMP-8 proteins and mRNA were detected in the conditioned media and cell lysates in 11 UM cell lines and three uveal melanoma cell lines cultured without serum, but not in media and cell lysates from other ocular resident cells or 12 malignant cell lines from other tissues, with exception of cutaneous melanoma cells. TNF-α moderately increased MMP-8 mRNA and protein levels in a dose- and time-dependent manner, accompanied by a significant increase of phosphorylated JNK1/2 and ERK1/2 in cell lysates. ERK1/2 (U0126) and JNK1/2 (SP600125) inhibitors significantly blocked TNF-α-induced and constitutive expression of MMP-8 in UM. This is the first report on the expression and secretion of MMP-8 by UM and uveal melanoma cells. The data suggest that UM may play a role in the remolding process and pathogenesis of inflammatory-related diseases in the eye via secretion of MMP-8.

Authors : Hu Dan-Ning, Rosen Richard B, Chan Chi-Chao, Yang Wei-En, Yang Shun-Fa,

(5) γ-tocotrienol inhibits the invasion and migration of human gastric cancer cells through downregulation of cyclooxygenase-2 expression.[TOP]

Pubmed ID :29901169
Publication Date : //
γ-tocotrienol (γ-T3), a tocotrienol isoform belonging to the vitamin E family, has been revealed to exert inhibitory effects on proliferation, migration and invasion in human gastric cancer cells. However, its precise mechanism of action is still unclear and needs to be further tested. Cyclooxygenase-2 (COX-2) is well known for its key role in promoting the migration and invasion abilities of human gastric cancer cells. In light of these data, our study aimed to validate whether the inhibitory actions of γ-T3 could be achieved by downregulation of COX-2 activity in vitro. In the present study, a Cell Counting Kit-8 (CCK-8) assay was performed to observe proliferation in human gastric cancer cells (SGC-7901 and MGC-803 cells), and wound healing and Transwell chamber assays were performed to detect migration and invasion. Western blot analyses were performed to analyse the relative expression of COX-2, matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) proteins, and enzyme-linked immunosorbent assays (ELISA) were used to determine the exocrine roles of MMP-2 and MMP-9. The results revealed that γ-T3 exerted significant inhibitory effects on proliferation, migration, invasion and COX-2 protein expression, as well as on exocrine functions of MMP-2 and MMP-9 in SGC-7901 and MGC-803 cells. Therefore, our results indicated that γ-T3 exerts inhibitory effects on migration and invasion, which may be mediated through downregulation of COX-2 expression in SGC-7901 and MGC-803 cells.

Authors : Zhang Ya Hui, Ma Ke, Liu Jia Ren, Wang Hai Xia, Tian Wen Xia, Tu Yue Hua, Sun Wen Guang,

(6) Circulating cathepsin B and D in pregnancy.[TOP]

Pubmed ID :29884105
Publication Date : //
The purpose of this prospective study was to investigate the changes in the circulating levels of cathepsin B and D in pregnancy. We obtained longitudinal cathepsin B and D levels in 76 healthy pregnant women in the first and third trimesters and compared these levels with 20 non-pregnant controls. The plasma levels of soluble cathepsin B and D were measured using an enzyme-linked immunosorbent assay kit. The cathepsin D concentrations in the third trimester were significantly higher than that in the first trimester (p < .001), and the cathepsin D levels in the first trimester were significantly lower than that in the non-pregnant controls (p = .002). However, there was no significant difference in the cathepsin B level throughout pregnancy compared to the non-pregnant controls. Our study is unique in evaluating the longitudinal changes in the cathepsin B and D levels in pregnancies without obstetric complications. The results implicate that changes in the levels of cathepsins might be essential in placentation. Therefore, molecular and genetic studies on cathepsin B and D are needed to understand the roles of these enzymes in pregnancy, thereby contributing to the understanding of placentation. Impact statement What is already known? Matrix metalloproteinases (MMP) have been widely studied, and their function is very important in the normal implantation process. The level of MMP-9 is known to increase throughout pregnancy, while the level of MMP-2 decreases in the first trimester. In addition to MMPs, other proteases are important for placental development; cathepsins B and D are two of the proteases that are involved in the normal placentation process. The function of cathepsin D is related to MMPs because this protease can activate MMPs either directly or indirectly. Nevertheless, the role of circulating cathepsins in pregnancy has not yet been fully elucidated. What do these results add? This study provides evidence, for the first time, that there are fluctuations of plasma cathepsin D level and there are no changes in the plasma cathepsin B level in a normal pregnancy. Moreover, we demonstrated that a cathepsin D level is significantly decreased in the first trimester compared to the non-pregnant controls, and that the level is markedly elevated in the third trimester. What are the implications of these findings for clinical practice and/or further research? Cathepsins B and D should be further studied locally in the placenta to explain the differences in the concentration of cathepsin D and no changes in cathepsin B, thereby exploring their exact roles.

Authors : Kim Ho Yeon, Baek Hye Sung,

(7) Direct determination of neonicotinoid insecticides in an analytically challenging crop such as Chinese chives using selective ELISAs.[TOP]

Pubmed ID :29869926
Publication Date : //
Easy-to-use commercial kit-based enzyme-linked immunosorbent assays (ELISAs) have been used to detect neonicotinoid dinotefuran, clothianidin and imidacloprid in Chinese chives, which are considered a troublesome matrix for chromatographic techniques. Based on their high water solubility, water was used as an extractant. Matrix interference could be avoided substantially just diluting sample extracts. Average recoveries of insecticides from spiked samples were 85-113%, with relative standard deviation of <15%. The concentrations of insecticides detected from the spiked samples with the proposed ELISA methods correlated well with those by the reference high-performance liquid chromatography (HPLC) method. The residues analyzed by the ELISA methods were consistently 1.24 times that found by the HPLC method, attributable to loss of analyte during sample clean-up for HPLC analyses. It was revealed that the ELISA methods can be applied easily to pesticide residue analysis in troublesome matrix such as Chinese chives.

Authors : Watanabe Eiki, Miyake Shiro,

(8) Modulation the crosstalk between tumor-associated macrophages and non-small cell lung cancer to inhibit tumor migration and invasion by ginsenoside Rh2.[TOP]

Pubmed ID :29783929
Publication Date : //
Tumor-associated macrophages (TAMs) play a critical role in modulating the tumor microenvironment and promote tumor metastases. Our studies have demonstrated that ginsenoside Rh2 (G-Rh2), a monomeric compound extracted from ginseng, is a promising anti-tumor agent in lung cancer cells. However, it remains unclear whetherG-Rh2 can modulate the differentiation of TAMs and its interaction with tumor microenvironment. In this study, we investigated how G-Rh2 regulates the phenotype of macrophages and affects the migration of non-small cell lung cancer (NSCLC) cells.

Authors : Li Honglin, Huang Nan, Zhu Weikang, Wu Jianchun, Yang Xiaohui, Teng Wenjing, Tian Jianhui, Fang Zhihong, Luo Yingbin, Chen Min, Li Yan,

(9) Regulation of fibroblast Fas expression by soluble and mechanical pro-fibrotic stimuli.[TOP]

Pubmed ID :29747634
Publication Date : //
Fibroblast apoptosis is a critical component of normal repair and the acquisition of an apoptosis-resistant phenotype contributes to the pathogenesis of fibrotic repair. Fibroblasts from fibrotic lungs of humans and mice demonstrate resistance to apoptosis induced by Fas-ligand and prior studies have shown that susceptibility to apoptosis is enhanced when Fas (CD95) expression is increased in these cells. Moreover, prior work shows that Fas expression in fibrotic lung fibroblasts is reduced by epigenetic silencing of the Fas promoter. However, the mechanisms by which microenvironmental stimuli such as TGF-β1 and substrate stiffness affect fibroblast Fas expression are not well understood.

Authors : Dodi Amos E, Ajayi Iyabode O, Chang Christine, Beard Meghan, Ashley Shanna L, Huang Steven K, Thannickal Victor J, Tschumperlin Daniel J, Sisson Thomas H, Horowitz Jeffrey C,

(10) [Preparation of rat uterine decellularized scaffold and extracellular matrix hydrogel].[TOP]

Pubmed ID :29745529
Publication Date : //
The chemical extraction method was used to prepare the rat uterine decellularized scaffolds, and to investigate the feasibility of preparing the extracellular matrix (ECM) hydrogel. The rat uterus were collected and extracted by 1%sodium dodecyl sulfate (SDS), 3% TritonX-100 and 4% sodium deoxycholate (SDC) in sequence. Scanning electron microscopy, histochemical staining and immunohistochemistry was used to assess the degree of decellularization of rat uterine scaffold. The prepared decellularized scaffold was digested with pepsin to obtain a uterine ECM hydrogel, and the protein content of ECM was determined by specific ELISA kit. Meanwhile, the mechanical characteristic of ECM hydrogel was measured. The results showed that the chemical extraction method can effectively remove the cells effectively in the rat uterine decellularized scaffold, with the ECM composition preserved completely. ECM hydrogel contains a large amount of ECM protein and shows a good stability, which provides a suitable supporting material for the reconstruction of endometrium .

Authors : Xu Jie, Jin Binghui, Zhao Yingzheng,