scholarly journals OPTIMIZED METHOD OF BOVINE PERICARDIUM DECELLULARIZATION FOR TISSUE ENGINEERING

2021 ◽  
Vol 74 (4) ◽  
pp. 815-820
Author(s):  
Nataliia V. Shchotkina ◽  
Anatoliy A. Sokol ◽  
Oleksandr Yu. Galkin ◽  
Glib I. Yemets ◽  
Liudmyla V. Dolinchuk ◽  
...  
Keyword(s):  
Cardiac Surgery ◽  
Human Fibroblasts ◽  
Molecular Genetic ◽  
Sodium Dodecyl ◽  
Laboratory Animals ◽  
Cross Linking ◽  
Prolonged Contact ◽  
Potential Possibility ◽  
High Degree

The aim: To investigate the effectiveness of using low concentrations of sodium dodecyl sulfate (SDS) and cross-linking with EDC/NHS in the decellularization process to create a potential bioimplant for cardiac surgery. Materials and methods: Pericardial sacs were derived from 12-18 months bulls. Tissue decellularization was performed by using 0.1% SDS with the following EDC/NHS cross-linking. The experiment included standard histological, microscopic, molecular genetic and biomechanical methods. Scaffold was tested in vitro for cytotoxicity and biocompatibility. Results: A high degree of extracellular decellularized matrix purification from cells and their components was shown. Structure-function properties remained similar to those or even improved after the decellularization. During prolonged contact of BP with human fibroblasts, no cytotoxic effect was observed. The biointegration of the scaffold in laboratory animals tissues was noted confirming the potential possibility of the implant use in cardiac surgery. Conclusions: Decellularization of BP by 0.1 % SDS with NHS/EDC cross-linking is promising in manufacturing of the tissue-engineered materials in cardiac surgery.

scholarly journals PROSPECTS FOR APPLICATION OF BOVINE PERICARDIAL SCAFFOLD FOR CARDIAL SURGERY

2020 ◽  
Vol 13 (6) ◽  
pp. 41-49
Author(s):  
A. A. Sokol ◽  
◽  
◽  
Keyword(s):  
Cytotoxic Effect ◽  
Molecular Genetic ◽  
Sodium Dodecyl ◽  
Bovine Pericardium ◽  
Laboratory Animals ◽  
Prolonged Contact ◽  
Potential Possibility ◽  
High Degree

The aim of the\* study was to estimate the properties of the scaffold obtained by decellularization of bovine pericardium with a 0.1% solution of sodium dodecyl sulfate. The experiment included standard histological, microscopic, molecular genetic, and biomechanical methods. Scaffold was tested in vitro for cytotoxicity and in vivo for biocompatibility. A high degree of removal of cells and their components from bovine pericardium-derived matrix was shown. Biomechanical characteristics of artificial scaffold were the same as those of the native pericardium. With prolonged contact, no cytotoxic effect on human cells was observed. The biointegration of the scaffold in laboratory animals tissues was noted, which confirms the potential possibility of the implant applicationin cardiac surgery.

scholarly journals MicroRNA-19b is a potential biomarker of increased myocardial collagen cross-linking in patients with aortic stenosis and heart failure

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Javier Beaumont ◽  
Begoña López ◽  
Susana Ravassa ◽  
Nerea Hermida ◽  
Gorka San José ◽  
...  
Keyword(s):  
Heart Failure ◽  
Aortic Stenosis ◽  
Severe Aortic Stenosis ◽  
Human Fibroblasts ◽  
Tissue Growth ◽  
Cross Linking ◽  
Collagen Network ◽  
Potential Biomarker ◽  
Myocardial Collagen

AbstractThis study analyzed the potential associations of 7 myocardial fibrosis-related microRNAs with the quality of the collagen network (e.g., the degree of collagen fibril cross-linking or CCL) and the enzyme lysyl oxidase (LOX) responsible for CCL in 28 patients with severe aortic stenosis (AS) of whom 46% had a diagnosis of chronic heart failure (HF). MicroRNA expression was analyzed in myocardial and blood samples. From the studied microRNAs only miR-19b presented a direct correlation (p < 0.05) between serum and myocardium. Compared to controls both myocardial and serum miR-19b were reduced (p < 0.01) in AS patients. In addition, miR-19b was reduced in the myocardium (p < 0.01) and serum (p < 0.05) of patients with HF compared to patients without HF. Myocardial and serum miR-19b were inversely correlated (p < 0.05) with LOX, CCL and LV stiffness in AS patients. Inin vitrostudies miR-19b inhibition increased (p < 0.05) connective tissue growth factor protein and LOX protein expression in human fibroblasts. In conclusion, decreased miR-19b may be involved in myocardial LOX up-regulation and excessive CCL, and consequently increased LV stiffness in AS patients, namely in those with HF. Serum miR-19b can be a biomarker of these alterations of the myocardial collagen network in AS patients, particularly in patients with HF.

Molecular cytogenetic analysis of wheat-alien hybrids and derivatives

2002 ◽  
Vol 50 (3) ◽  
pp. 303-311 ◽  
Author(s):  
M. Molnár-Láng ◽  
G Linc ◽  
E. D. Nagy ◽  
Keyword(s):  
Ssr Markers ◽  
Molecular Genetic ◽  
Colchicine Treatment ◽  
Translocation Chromosome ◽  
Chromosome Polymorphism ◽  
Translocation Breakpoints ◽  
Mother Cells ◽  
High Degree

New wheat × barley, wheat × Aegilops biuncialis and wheat × rye hybrids were produced with the aim of alien gene transfer from these species into wheat. Amphiploids were produced with the help of colchicine treatment from the last two combinations. The new wheat × barley hybrids were multiplied in tissue culture because of the high degree of sterility and then pollinated with wheat to obtain backcross progenies. Wheat-barley chromosome pairing was detected using genomic in situ hybridization (GISH) in two combinations (Mv9 kr1 × Igri, Asakazekomugi × Manas). In vitro conditions caused an increase in chromosome arm association frequency in both combinations and in fertility in some regenerants. Five wheat-barley translocations were produced in a wheat background and characterized through the combination of cytogenetic and molecular genetic approaches (GISH, FISH and SSR markers). The following translocations were identified: 2DS.2DL-1HS, 3HS.3BL, 6BS.6BL-4HL, 4D-5HS and 7DL.7DS-5HS. Physical mapping of the SSR markers on chromosomes 1H and 5H was carried out using the intragenomic and interspecific translocation breakpoints and the centromere as physical landmarks.  Disomic wheat-Aegilops biuncialis additions were produced after backcrossing the wheat-Ae. biuncialis amphiploids. Fluorescence in situ hybridization (FISH) was carried out using two repetitive DNA clones (pSc119.2 and pAs1) on Ae. biuncialis and its two diploid progenitor species to detect chromosome polymorphism. The 7M and 3M disomic chromosome additions were selected and five more lines still need to be characterized.  The octoploid triticale (Mv9 kr1 × Lovászpatonai) produced in Martonvásár was crossed with a 1RS.1BL wheat cultivar Matador. GISH analysis detected pairing between the 1RS arm of the translocation chromosome and that of Lovászpatonai rye in 32 % of the pollen mother cells, making it possible to select recombinants from this combination. The new recombinants between the 1RS of Petkus and the 1RS of Lovászpatonai rye cultivars are being analysed with the help of microsatellite markers.

scholarly journals Effect of tea root-derived proanthocyanidin fractions on protection of dentin collagen

2019 ◽  
Vol 48 (5) ◽  
pp. 030006051989130
Author(s):  
Honglin Yang ◽  
Bingqing Xie ◽  
Yue Wang ◽  
Yayun Cui ◽  
Hui Yang ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Butyl Alcohol ◽  
Degree Of Polymerization ◽  
Cross Linking ◽  
Protective Mechanisms ◽  
Dentin Collagen ◽  
And Function ◽  
High Degree ◽  
Collagen Cross Linking

Objectives Proanthocyanidins (PAs) have been widely used as effective agents for dentin collagen cross-linking to enhance the biomechanics and biostability of dentin in vitro. However, the effects and protective mechanisms of various tea root-derived PA components on dentin remain undefined. This study evaluated the effects of these tea root-derived PA components on dentin biomechanics and biostability. Methods In this study, ethyl acetate and n-butyl alcohol were used to extract PAs with different degrees of polymerization from tea roots; the effects of these PA extracts on dentin were evaluated. Results Dentin was treated with glutaraldehyde, ethyl acetate, n-butyl alcohol, or water. PAs with a high degree of polymerization, extracted using n-butyl alcohol, were able to more effectively improve dentin collagen cross-linking, increase resistance to bacterial collagenase digestion, and enhance dentin elasticity, relative to treatment with glutaraldehyde or PAs with a low degree of polymerization (extracted using ethyl acetate). Additionally, treatment with aqueous extract of tea roots was detrimental to dentin stability and function. Conclusions PAs with a high degree of polymerization were effective for dentin protection and restoration in vitro, suggesting clinical treatment potential for tea root-derived PAs.

scholarly journals Exocrine pancreas transcription factor 1 binds to a bipartite enhancer element and activates transcription of acinar genes.

1991 ◽  
Vol 11 (10) ◽  
pp. 4985-4997 ◽  
Author(s):  
S L Weinrich ◽  
A Meister ◽  
W J Rutter
Keyword(s):  
Transcription Factor ◽  
Exocrine Pancreas ◽  
Sodium Dodecyl ◽  
Binding Activity ◽  
Cross Linking ◽  
Major Protein ◽  
Enhancer Element ◽  
Transcription Factor 1

Exocrine pancreas (XP) enhancers, which contain a conserved core sequence, are active only in XP cells. A core enhancer-binding activity also appears to be restricted to XP nuclei. Here we describe the properties of a factor, purified approximately 100,000-fold from pancreas nuclei, which displays core enhancer-binding activity. It is not identical to previously characterized factors and is termed exocrine pancreas transcription factor 1 (XPF-1). In the highly purified preparation, only a single major protein of 60 kDa was detected by silver staining on sodium dodecyl sulfate-gels and by UV cross-linking. XPF-1 binds to the core enhancer of all tested XP genes and not to a mutant sequence which is inactive in vivo. High-affinity binding sites are bipartite. The results of competition binding and UV-cross-linking assays suggest that XPF-1 interacts with both motifs. XPF-1 selectively stimulates transcription of core enhancer templates in an in vitro transcription system. We hypothesize that XPF-1 plays a role in activation of the transcription of XP-specific genes.

scholarly journals Biosynthesis of the first component of complement by human fibroblasts

1977 ◽  
Vol 167 (3) ◽  
pp. 647-660 ◽  
Author(s):  
Kenneth B. M. Reid ◽  
Ellen Solomon
Keyword(s):  
Human Serum ◽  
Cell Lines ◽  
De Novo ◽  
Culture Media ◽  
Human Fibroblasts ◽  
Sodium Dodecyl ◽  
Fibroblast Cell ◽  
Haemolytic Activity ◽  
Fibroblast Cell Lines

1. Haemolytic activity corresponding to that of the first component of complement (C1) was synthesized and secreted by all nine human fibroblast cell lines examined. No activity was found in the culture media of a variety of other human cell lines. 2. The component-C1 haemolytic activity secreted by the fibroblast lines behaved in an identical manner, in most respects, with that of the component-C1 haemolytic activity of human serum. The component-C1 haemolytic activity secreted by fibroblasts, however, was less susceptible to inhibition by rabbit fragment F(ab′)2 anti-(human subcomponent C1q) than was the component-C1 haemolytic activity of human serum. 3. Biosynthesis of fibroblast component-C1 haemolytic activity was inhibited by the presence of cycloheximide and regained on its removal. 4. Incorporation of radioactivity into proteins secreted by the fibroblasts and release of component-C1 haemolytic activity by the fibroblasts both increased in a linear manner until several days after the cultures had reached a state of confluent growth. 5. Radioactivity was incorporated into subcomponents C1q, C1r and C1s, as judged by the formation of specific immunoprecipitates and by absorption with immune aggregates. 6. The immunoprecipitates formed by using antisera against subcomponents C1r and C1s were run on polyacrylamide gels in sodium dodecyl sulphate, and this provided convincing physiochemical evidence for the biosynthesis of these subcomponents de novo. 7. The results obtained with immunoprecipitates formed by using anti-(subcomponent C1q) suggest that subcomponent C1q may be synthesized and secreted by fibroblast cell lines in vitro, in a form with a higher molecular weight than that of subcomponent C1q which is isolated by conventional techniques of protein fractionation from fresh serum.

Exocrine pancreas transcription factor 1 binds to a bipartite enhancer element and activates transcription of acinar genes

1991 ◽  
Vol 11 (10) ◽  
pp. 4985-4997
Author(s):  
S L Weinrich ◽  
A Meister ◽  
W J Rutter
Keyword(s):  
Transcription Factor ◽  
Exocrine Pancreas ◽  
Sodium Dodecyl ◽  
Binding Activity ◽  
Cross Linking ◽  
Major Protein ◽  
Enhancer Element ◽  
Transcription Factor 1

Exocrine pancreas (XP) enhancers, which contain a conserved core sequence, are active only in XP cells. A core enhancer-binding activity also appears to be restricted to XP nuclei. Here we describe the properties of a factor, purified approximately 100,000-fold from pancreas nuclei, which displays core enhancer-binding activity. It is not identical to previously characterized factors and is termed exocrine pancreas transcription factor 1 (XPF-1). In the highly purified preparation, only a single major protein of 60 kDa was detected by silver staining on sodium dodecyl sulfate-gels and by UV cross-linking. XPF-1 binds to the core enhancer of all tested XP genes and not to a mutant sequence which is inactive in vivo. High-affinity binding sites are bipartite. The results of competition binding and UV-cross-linking assays suggest that XPF-1 interacts with both motifs. XPF-1 selectively stimulates transcription of core enhancer templates in an in vitro transcription system. We hypothesize that XPF-1 plays a role in activation of the transcription of XP-specific genes.

Anti-Diabetic Activity of a Mineraloid Isolate, in vitro and in Genetically Diabetic Mice

2011 ◽  
Vol 81 (1) ◽  
pp. 34-42 ◽  
Author(s):  
Joel Deneau ◽  
Taufeeq Ahmed ◽  
Roger Blotsky ◽  
Krzysztof Bojanowski
Keyword(s):  
Dna Microarrays ◽  
Human Fibroblasts ◽  
Diabetic Animal ◽  
In Vitro Tests ◽  
Diabetic Mice ◽  
Fossil Material ◽  
Expression Of Genes ◽  
Enhanced Expression ◽  
Genetically Diabetic Mice

Type II diabetes is a metabolic disease mediated through multiple molecular pathways. Here, we report anti-diabetic effect of a standardized isolate from a fossil material - a mineraloid leonardite - in in vitro tests and in genetically diabetic mice. The mineraloid isolate stimulated mitochondrial metabolism in human fibroblasts and this stimulation correlated with enhanced expression of genes coding for mitochondrial proteins such as ATP synthases and ribosomal protein precursors, as measured by DNA microarrays. In the diabetic animal model, consumption of the Totala isolate resulted in decreased weight gain, blood glucose, and glycated hemoglobin. To our best knowledge, this is the first description ever of a fossil material having anti-diabetic activity in pre-clinical models.

Tridegin, a Novel Peptidic Inhibitor of Factor XIIIa from the Leech, Haementeria ghilianii, Enhances Fibrinolysis In Vitro

1997 ◽  
Vol 77 (05) ◽  
pp. 0959-0963 ◽  
Author(s):  
Lisa Seale ◽  
Sarah Finney ◽  
Roy T Sawyer ◽  
Robert B Wallis
Keyword(s):  
Potent Inhibitor ◽  
Platelet Rich Plasma ◽  
Factor Xiiia ◽  
Cross Linking ◽  
Fibrinolytic Enzymes ◽  
Small Polypeptide ◽  
Tissue Plasminogen ◽  
Protein Cross Linking

SummaryTridegin is a potent inhibitor of factor Xllla from the leech, Haementeria ghilianii, which inhibits protein cross-linking. It modifies plasmin-mediated fibrin degradation as shown by the absence of D-dimer and approximately halves the time for fibrinolysis. Plasma clots formed in the presence of Tridegin lyse more rapidly when either streptokinase, tissue plasminogen activator or hementin is added 2 h after clot formation. The effect of Tridegin is markedly increased if clots are formed from platelet-rich plasma. Platelet-rich plasma clots are lysed much more slowly by the fibrinolytic enzymes used and if Tridegin is present, the rate of lysis returns almost to that of platelet- free clots. These studies indicate the important role of platelets in conferring resistance to commonly used fibrinolytic enzymes and suggest that protein cross-linking is an important step in this effect. Moreover they indicate that Tridegin, a small polypeptide, may have potential as an adjunct to thrombolytic therapy.

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