Matrix effects
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Due to high biocompatibility, miniaturization, optical transparency and low production cost together with high radiation hardness the diamond-based sensors are considered promising for radiation medicine and biomedicine in general. Here we present detection of fibroblast cell culture properties by nanocrystalline diamond solution-gated field-effect transistors (SG-FET), including effects of gamma irradiation. We show that blank nanocrystalline diamond field-effect biosensors are stable at least up to 300 Gy of γ irradiation. On the other hand, gate current of the diamond SG-FET biosensors with fibroblastic cells increases exponentially over an order of magnitude with increasing radiation dose. Extracellular matrix (ECM) formation is also detected and analyzed by correlation of electronic sensor data with optical, atomic force, fluorescence, and scanning electron microscopies.
- Klíčová slova
- Atomic force microscopy, Biosensors, Cells, Diamond thin films, Field-effect transistors, Gamma irradiation, Proteins,
- MeSH
- biosenzitivní techniky * MeSH
- diamant * MeSH
- extracelulární matrix MeSH
- fibroblasty MeSH
- mikroskopie elektronová rastrovací MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- diamant * MeSH
RATIONALE: Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is used for the fast qualitative and quantitative analysis of phosphatidylcholines (PC). Fatty acyl chain lengths and the number of double bonds (DB) affect relative responses of PC; hence the determination of correction factors of individual PC is important for the accurate quantitation. The signal intensity in MALDI-MS strongly depends on the matrix; therefore, the following matrices typically used in lipidomics are studied in the present work: 2,5-dihydroxybenzoic acid (DHB), 1,5-diaminonaphthalene (DAN) and 9-aminoacridine (9AA). METHODS: Series of PC with various fatty acyl chain lengths are synthesized for this study. PC concentrations over two orders of magnitude are studied with MALDI-MS. These experiments provide sets of calibration curves for each of the synthesized PC and the further analysis of parameters of calibration curves is performed. RESULTS: Correction factors for PC decrease with increasing fatty acyl chain length for all matrices. These dependences are steeper for unsaturated PC than for saturated ones. MALDI matrices also have a significant effect on this dependence. The weakest dependence on fatty acyl chain length is found for saturated PC in 9AA. In the case of the other matrices, the effect of fatty acyl chain length on the response is essential for both saturated and unsaturated PC. Calibration curves and parameters of calibration curves for both saturated and monounsaturated PC are fitted by a linear function with regression coefficients decreasing in the order 9AA > DAN > DHB. CONCLUSIONS: Differences in relative responses for PC in MALDI-MS measurements must be taken into account for accurate quantitation. Parameters of calibration curves can be used for the determination of PC concentrations using a single internal standard (IS). This method gives good results for the 9AA matrix, but the reproducibility of measurements for the DHB and DAN matrices is lower and the method can be used for a rough estimation only. These matrices are less convenient for the quantitation of PC.
- MeSH
- fosfatidylcholiny krev chemie MeSH
- lidé MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- fosfatidylcholiny MeSH
A significant number of myocardial diseases are accompanied by increased synthesis and degradation of the extracellular matrix (ECM) as well as by changed maturation and incorporation of ECM components. Important groups of enzymes responsible for both normal and pathological processes in ECM remodeling are matrix metaloproteinases (MMPs). These enzymes share a relatively conserved structure with a number of identifiable modules linked to their specific functions. The most important function of MMPs is the ability to cleave various ECM components; including such rigid molecules as fibrillar collagen molecules. The amount and activity of MMPs in cardiac tissue are regulated by a range of activating and inhibiting processes. Although MMPs play multifarious roles in many myocardial diseases, here we have focused on their function in ischemic cardiac tissue, dilated cardiomyopathy and hypertrophied cardiac tissue. The inhibition of MMPs by means of synthetic inhibitors seems to be a promising strategy in cardiac disease treatment. Their effects on diseased cardiac tissue have been successfully tested in several experimental studies.
- MeSH
- extracelulární matrix metabolismus MeSH
- inhibitory matrixových metaloproteinas MeSH
- kardiomyopatie metabolismus MeSH
- lidé MeSH
- matrixové metaloproteinasy metabolismus MeSH
- myokard metabolismus MeSH
- remodelace komor MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- inhibitory matrixových metaloproteinas MeSH
- matrixové metaloproteinasy MeSH
BACKGROUND/AIMS: Podocytes are typically cultured on collagen I; however, collagen I is absent from healthy glomerular basement membranes. Erythropoietin (EPO) is thought to protect podocytes in vivo. Here, we studied how various types of extracellular matrix (ECM) proteins and EPO affect podocytes in culture. METHODS: Primary rat podocytes were replated on collagen I, collagen IV, whole ECM extract, laminin, or bare plastic. Cellular adhesion (8 hours after plating), proliferation (5 days, 10 % serum), and resistance to serum deprivation (3 days, 0.5 % serum) were assessed. BrdU incorporation and expression of podocyte-specific markers were employed as measures of cellular proliferation and differentiation, respectively. qPCR was used to verify expression of EPO receptor in cultured podocytes. RESULTS: Cellular adhesion was similar on all ECM proteins and unaffected by EPO. Proliferation was accelerated by laminin and the ECM extract, but the final cell density was similar on all ECM surfaces. Collagen IV supported the serum-deprived cells better than the other ECM proteins. EPO (2-20 ng/ml) improved viability of serum-deprived podocytes on collagen I, collagen IV, and ECM, but not on laminin or bare plastic. The cells expressed mRNA for EPO receptor. CONCLUSION: The physiological ECM proteins are more supportive of primary podocytic cultures compared with collagen I. The protective effects of EPO during serum deprivation are modulated by the cultivation surface.
- MeSH
- barvicí látky MeSH
- erythropoetin farmakologie MeSH
- extracelulární matrix - proteiny fyziologie MeSH
- extracelulární matrix účinky léků metabolismus MeSH
- glomerulus účinky léků MeSH
- krysa rodu Rattus MeSH
- kultivované buňky MeSH
- podocyty účinky léků MeSH
- primární buněčná kultura MeSH
- receptory erythropoetinu biosyntéza účinky léků MeSH
- rekombinantní proteiny farmakologie MeSH
- tetrazoliové soli MeSH
- thiazoly MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- barvicí látky MeSH
- erythropoetin MeSH
- extracelulární matrix - proteiny MeSH
- receptory erythropoetinu MeSH
- rekombinantní proteiny MeSH
- tetrazoliové soli MeSH
- thiazoly MeSH
- thiazolyl blue MeSH Prohlížeč
Lithium 2,5-dihydroxybenzoate (LiDHB) is shown to be a very effective matrix for matrix-assisted laser desorption/ionization (MALDI) analysis of nonpolar long-chain lipids, hydrocarbons and polymers. Under standard desorption and ionization conditions using a conventional nitrogen UV laser (337 nm), hydrocarbons (C(24)-C(40)), diverse lipids (triglycerides, diglycerides, wax esters from leaves) and saturated polymers are effectively lithiated providing [M+Li](+) ions. The formation of lithiated hydrocarbons is not accompanied by an elimination of hydrogen or other fragmentation reactions and, due to the relatively simple isotopic distribution of lithium, seems to be more useable for analysis of hydrocarbon mixtures than the previously used silver cationization agents. The mass calibration can be conveniently performed either externally or internally using poly(ethylene glycol) commercial standards.
- MeSH
- gentisáty * MeSH
- glyceridy analýza chemie MeSH
- lipidy analýza chemie MeSH
- listy rostlin chemie MeSH
- lithium * MeSH
- molekulová hmotnost MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- uhlovodíky analýza chemie MeSH
- vosky chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 2,5-dihydroxybenzoic acid MeSH Prohlížeč
- carnauba wax MeSH Prohlížeč
- gentisáty * MeSH
- glyceridy MeSH
- lipidy MeSH
- lithium * MeSH
- uhlovodíky MeSH
- vosky MeSH
BACKGROUND: Clinical outcome after intracerebral hemorrhage (ICH) remains poor. Recent trials in ICH, focusing on hematoma reduction, have not yielded significant clinical improvement. The modulation of matrix metalloproteinase (MMP)-9 may represent a potential therapeutic target for reducing perihematomal edema (PHE) and improving clinical outcome. METHODS: We searched Cochrane Library, Ovid/Medline, and PubMed databases using combinations of the following MeSH search terms: "intracerebral hemorrhage," "matrix metalloproteinase," "minocycline," "inhibition," and "neuroprotection". RESULTS: MMP-9 levels in animal models have largely shown detrimental correlations with mortality, clinical outcome, hematoma volume, and PHE. Animal models and clinical studies have established a timeline for MMP-9 expression and corresponding PHE that include an initial peak on days 1-3 and a secondary peak on day 7. Clinical studies evaluating MMP-9 levels in the acute phase (days 1-3) and subacute phase (day 7) of ICH suggest that MMP-9 may be detrimental in the acute phase through destruction of basal lamina, activation of vascular endothelial growth factor, and activation of apoptosis but assist in recovery in the subacute phase through angiogenesis. CONCLUSIONS: MMP-9 inhibition represents a potentially effective target for neuroprotection in ICH. However, as a ubiquitous protein, the inhibition of pathologic processes must be balanced against the preservation of neuroprotective angiogenesis. As the opposing roles of MMP-9 may have similar mechanisms, the most important factor may be the timing of MMP-9 inhibition. Further studies are necessary to delineate these mechanisms and their temporal relationship.
- Klíčová slova
- Matrix metalloproteinase, blood brain barrier, gelatinase B, intracerebral hemorrhage, neuroprotection, perihematomal edema,
- MeSH
- apoptóza účinky léků MeSH
- časové faktory MeSH
- cerebrální krvácení komplikace farmakoterapie metabolismus mortalita patologie MeSH
- edém mozku farmakoterapie metabolismus mortalita MeSH
- hematom farmakoterapie metabolismus mortalita MeSH
- inhibitory matrixových metaloproteinas terapeutické užití MeSH
- lidé MeSH
- matrixová metaloproteinasa 9 metabolismus MeSH
- neuroprotektivní látky aplikace a dávkování MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- metaanalýza MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- inhibitory matrixových metaloproteinas MeSH
- matrixová metaloproteinasa 9 MeSH
- neuroprotektivní látky MeSH
Exercise induced bone response although established, little is known about the molecular components that mediate bone response to mechanical loading (ML). In our recent QTL study, we identified one such possible molecular component responding to ML: cartilage oligomeric matrix protein (COMP). To address the COMP role in mediating ML effects on bone formation, COMP expression was evaluated as a function of duration and age in response to ML in female B6 mice. A 9N load was applied using a four-point bending device at 2Hz frequency for 36 cycles, once per day for 2-, 4- and 12-days on the right tibia. The left tibia was used as an internal control. Loading caused an increase in COMP expression by 1.3-, 2- and 4-fold respectively after 2-, 4- and 12-days of loading. This increase was also seen in 16 and 36-week old mice. Based on these findings, we next used COMP knockout (KO) mice to evaluate the cause and effect relationship. Quantitative analysis revealed 2 weeks of ML induced changes in vBMD and bone size in the KO mice (5.9 % and 21 % vs. unloaded bones) was not significantly different from control mice (7 % and 24 % vs. unloaded bones). Our results imply that COMP is not a key upstream mediator of the anabolic effects of ML on the skeleton.
- MeSH
- extracelulární matrix - proteiny genetika metabolismus MeSH
- glykoproteiny genetika metabolismus MeSH
- kosti a kostní tkáň fyziologie MeSH
- lokus kvantitativního znaku MeSH
- matriliny MeSH
- mechanický stres MeSH
- myši knockoutované MeSH
- myši MeSH
- osteogeneze fyziologie MeSH
- zatížení muskuloskeletálního systému fyziologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Názvy látek
- extracelulární matrix - proteiny MeSH
- glykoproteiny MeSH
- Matn1 protein, mouse MeSH Prohlížeč
- matriliny MeSH
INTRODUCTION: Preeclampsia is a life-threatening condition for the mother and foetus. Globally, it is dia-gnosed in 10 mil. women every year, which accounts for 3% to 8% of all pregnancies. Currently there is no proven effective treatment for preeclampsia. The aforesaid text actualises the issue of predicting this complication. To determine the prognostic significance of matrix metalloproteinases-2 and -9 levels as early markers of preeclampsia, the present prospective study was conducted. MATERIALS AND METHODS: The levels of matrix metalloproteinases-2 and -9 were assessed in 72 patients. Thirty-four of them subsequently developed preeclampsia during pregnancy (20 patients with moderate preeclampsia, 14 patients with severe preeclampsia), and constituted the basic group; 38 patients made up the control group. RESULTS: In pregnant women with the subsequent development of preeclampsia, the level of matrix metalloproteinase-2 at 11-13 weeks of gestation was 155 ± 73.4 ng/mL and significantly exceeded its level in pregnant women without hypertensive disorders - 75.0 ± 32.8 ng/mL. The study conducted demonstrates a significantly lower concentration of matrix metalloproteinase-9 in pregnant women with preeclampsia compared to the control - 749 ± 296 ng/mL and 1,667 ± 552 ng/mL (P < 0.001). The performed research figures that in the first trimester, the cut-off value of matrix metalloproteinase-2 for predicting the development of preeclampsia is 102 ng/mL (sensitivity 88.24% and specificity 82.76%). For matrix metalloproteinase-9, a level of 980 ng/mL in the first trimester predicts the development of preeclampsia with a sensitivity of 85.29% and a specificity of 84.48%. CONCLUSION: The study established the cut-off values of matrix metalloproteinases-2 and -9 for predicting the development of preeclampsia in the first trimester.
- Klíčová slova
- matrix metalloproteinases, matrix metaloproteinases, prediction of preeclampsia, preeclampsia, pregnancy,
- MeSH
- biologické markery MeSH
- lidé MeSH
- matrixová metaloproteinasa 2 * MeSH
- matrixová metaloproteinasa 9 MeSH
- preeklampsie * diagnóza MeSH
- prospektivní studie MeSH
- první trimestr těhotenství MeSH
- těhotenství MeSH
- Check Tag
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- matrixová metaloproteinasa 2 * MeSH
- matrixová metaloproteinasa 9 MeSH
PURPOSE: Matrix metalloproteinases (MMPs) are postulated to be involved in the development of retinal angiogenesis through the regulation of extracellular matrix. The objective of the present study was to test for a possible association of five single nucleotide polymorphisms (SNPs) in the MMP-2 gene and two polymorphisms in the MMP-9 gene with proliferative diabetic retinopathy (PDR) and to determine their plasma levels. METHODS: The study comprised 490 Caucasian participants, who were divided into three groups: diabetics with PDR, diabetics without PDR, and nondiabetics. Genotypes were detected by polymerase chain reactions followed by restriction analyses with specific endonucleases and their frequencies determined. Plasma levels of MMP-2 and MMP-9 proteins were analyzed by ELISA. RESULTS: Neither MMP-2 SNPs nor MMP-9 SNPs revealed significant association with PDR in single-locus comparisons; similarly, MMP-2 haplotype frequencies did not differ notably between groups, although the C-allele of the -1306C/T polymorphism and the C-allele containing haplotype (CGCG) in MMP-2 exhibited marginally significant association with PDR in males (p<0.05, p(corr)=NS). Both MMP-2 and MMP-9 plasma levels showed statistically significant differences among the studied groups (p<0.001 and p=0.001, respectively) with highest levels in the PDR group. MMP-2 plasma levels were markedly higher in carriers of either the -1306CC and -1306CT genotypes and (p=0.009) or CGCG haplotype (p=0.043). CONCLUSIONS: These findings indicate that genotype- and haplotype-specific effects on MMP-2 expression corresponding with its plasma levels may contribute to the susceptibility to PDR.
- MeSH
- diabetická retinopatie enzymologie genetika patologie MeSH
- dospělí MeSH
- genetická variace MeSH
- haplotypy MeSH
- lidé středního věku MeSH
- lidé MeSH
- matrixová metaloproteinasa 2 krev genetika MeSH
- matrixová metaloproteinasa 9 krev genetika MeSH
- pohlavní dimorfismus MeSH
- progrese nemoci MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- matrixová metaloproteinasa 2 MeSH
- matrixová metaloproteinasa 9 MeSH
The extracellular matrix (ECM) consists of proteins, glycosaminoglycans and glycoproteins, that support the dynamic interactions between cells, including intercellular communication, cell attachment, cell differentiation, cell growth and migration. As such, the ECM represents an essential and very sensitive system within the tissue microenvironment that is involved in processes such as tissue regeneration and carcinogenesis. The aim of the present review is to evaluate its diversity through Ca(2+) signaling and its role in muscle cell function. Here, we discuss some methodological approaches dissecting Ca(2+) handling mechanisms in myogenic and non-myogenic cells, e.g. the importance of Ca(2+) and calpains in muscle dystrophy. We also consider the reconstruction of skeletal muscle by colonization of decellularized ECM with muscle-derived cells isolated from skeletal muscle. Therefore, it is necessary to establish new methodological procedures based on Ca(2+) signaling in skeletal muscle cells and their effect on ECM homeostasis, allowing the monitoring of skeletal muscle reconstruction and organ repair.
