scholarly journals Validation of chicken-specific pro-angiogenic growth factors three-dimensional structures and analysis of their role in high molecular weight heparin-induced angiogenesis

2021 ◽  
Author(s):  
Reji Manjunathan ◽  
Vijayalakshmi Periyaswami ◽  
Malathi Ragunathan
Keyword(s):  
Molecular Weight ◽  
Growth Factors ◽  
High Molecular Weight ◽  
Binding Affinity ◽  
In Silico ◽  
Three Dimensional ◽  
Angiogenic Growth Factors ◽  
Transcription Level ◽  
Physiological Conditions ◽  
Micromolar Concentration

Abstract Background - High-molecular weight heparin (HMWH), a molecule which is extensively in use as an anticoagulant shows concentration-dependent angiogenic and anti-angiogenic potential. Based on the concentration, HMWH can bind with both angiogenic and anti-angiogenic factors and exerts diverse effect. Our earlier data suggested that HMWH (15 kDa) can induce concentration-dependent neovascularization on chicken chorioallantoic membrane (CAM). The diffusion pattern of HMWH through various layers of CAM supports its internalized action with the various cellular components of angiogenesis. So far, no studies have reported the interactive potential of HMWH with various pro-angiogenic growth factors under physiological conditions. Hence, we aimed to find the transcription level interaction of HMWH with major pro-angiogenic growth factors. In connection to the research, for the first time, we validated the three-dimensional structures of chicken-specific pro-angiogenic growth factors such as FGF2, MMP2, MMP9, NOS3, VEGF A, and VEGF C to find the binding affinity of HMWH with the core-functional units of these growth factors. Methods - CAMs are incubated with 50, 100, and 150 µM concentration of HMWH. Changes in the transcription level of specified pro-angiogenic growth factors are analyzed by semi-PCR method. The functional aspects of these molecules are identified with zymogram and immunohistochemical approaches. Scanning electron microscopic technique is applied to find the morphological changes on CAM under HMWH incubation. Three-dimensional structure validation and molecular docking are performed using the SWISS-MODEL web server and AutoDock vina-PyRx software version 8.0. Results - HMWH can enhance the transcription level of major pro-angiogenic growth factors with a significant impact on FGF2 and MMP2 under 100 µM concentration. The in-silico analysis reveals that HMWH shows a higher binding affinity with FGF2 followed by MMP2, MMP9, NOS3, VEGF A, and VEGF C, respectively. Conclusion - The combined results from the experimental and in-silico analysis reveal that HMWH can interact with pro-angiogenic growth factors under micromolar concentration in physiological conditions while inducing angiogenesis. This observation further supports the therapeutic benefits of HMWH as an angiogenic factor under micromolar concentration.

scholarly journals Identification of B-cell growth factors (interleukin-14; high molecular weight-B-cell growth factors) in effusion fluids from patients with aggressive B-cell lymphomas

Blood ◽  
1995 ◽  
Vol 86 (1) ◽  
pp. 283-293 ◽  
Author(s):  
R Ford ◽  
A Tamayo ◽  
B Martin ◽  
K Niu ◽  
K Claypool ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Growth Factors ◽  
Cell Growth ◽  
B Cell ◽  
High Molecular Weight ◽  
Large Cell ◽  
Neoplastic Cell ◽  
Western Blots

The molecular basis of neoplastic B-cell growth is complex and poorly understood. Cytokines have been postulated to contribute to neoplastic cell growth, and many in vitro studies have confirmed this prediction, but little is known about the in vivo role of these growth factors. We have examined the production of interleukin-14 (IL-14) (high molecular weight [HMW], B-cell growth factor [BCGF]) by aggressive intermediate (diffuse large cell) lymphomas of the B-cell type non-Hodgkin's lymphoma (NHL-B) in four patients with lymphomatous effusions. In these studies, IL-14 was detected in the effusion fluids by Western blots and IL-14 mRNA was constitutively expressed in the freshly isolated lymphoma cells that also expressed the receptor for IL-14 (IL14R). Lymphoma B cells placed at low serum and cell density proliferated in vitro to either purified IL-14 or IL-14 derived from effusion fluids. Antibodies to IL-14 removed the growth-stimulating cytokine(s) from the effusions. Cell lines developed from these patients produced IL-14 in vitro and antisense oligos to IL-14 blocked their growth in vitro. Thus, autocrine or paracrine production of IL-14 may play a significant role in the rapid proliferation of aggressive NHL-B. Interrupting this pathway could be a useful goal of therapy for patients resistant to conventional chemotherapy.

scholarly journals STUDI MOLECULAR DOCKING SENYAWA GOLONGAN FLAVONOL SEBAGAI ANTIBAKTERI

2018 ◽  
Vol 1 (2) ◽  
pp. 20-27
Author(s):  
Isna Wardaniati ◽  
Muhammad Azhari Herli
Keyword(s):  
Molecular Docking ◽  
Binding Energy ◽  
Binding Affinity ◽  
Bioactive Compounds ◽  
In Silico ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Three Dimensional Structure ◽  
Receptor Interactions

In this paper we studied the bioactive compounds of Flavonol-D-alanil D-alanin dekarboksipeptidase receptor interactions In silico. First, prepared three dimensional structure of D-alanil D-alanin dekarboksipeptidase as receptor. Preparation of fourth bioactive compounds of flavonol which will be as ligands, klokasilin and D-alanil D-alanin as a comparison. The fourth bioactive compounds of flavonol, klokasilin and D-alanil D-alanin were docked with D-alanil D-alanin dekarboksipeptidase until energy values were obtained. The fourth bioactive compounds of flavonol had lesser binding energy values than D-alanil D-alanin, Quercitrine and rutin also predicted to have greater binding energy and binding affinity than klokasilin (antibiotic) and D-alanil D-alanin (nature ligand).

scholarly journals Clinical importance of proteinuria in diagnostic strategy for kidney diseases

2002 ◽  
Vol 21 (3) ◽  
pp. 291-295 ◽  
Author(s):  
Dejan Petrovic ◽  
Radmila Obrenovic ◽  
Nada Majkic-Singh ◽  
Mileta Poskurica ◽  
Biljana Stojimirovic
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Immunoglobulin G ◽  
Total Protein ◽  
Plasma Proteins ◽  
Kidney Diseases ◽  
Clinical Importance ◽  
Basal Membrane ◽  
Diagnostic Strategy ◽  
Physiological Conditions

Basal glomerular membrane represents mechanical and electrical obstacle for passing of plasma proteins. In physiological conditions only plasma proteins of low molecule weight are completely filtered through basal membrane. Due to damages of basal glomerular membrane there is increase in filtration of plasma protein of middle and high molecular weight. Depending on etiology of proteinuria it can be prerenal, renal and postrenal. By analyzing albumin a1-microglobulin, immunoglobulin G and a2-macroglobulin, together with total protein in urine, it is possible to detect and differentiate causes of prerenal, glomerular, tubular and postrenal proteinuria. The adequate and early differentiation of proteinuria is of an immense diagnostic and therapeutic importance.

A Modified Model of Type 2B von Willebrand Disease: Taking ADAMTS13-Mediated Cleavage out of the Equation

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 23-23
Author(s):  
Mia Golder ◽  
Cynthia M. Pruss ◽  
Kate Sponagle ◽  
Carol Hegadorn ◽  
Erin Burnett ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Binding Affinity ◽  
Full Range ◽  
Von Willebrand Disease ◽  
Platelet Glycoprotein ◽  
Platelet Counts ◽  
Platelet Binding ◽  
Von Willebrand ◽  
High Molecular Weight Multimers

Abstract Abstract 23 Introduction: Type 2B von Willebrand disease (2B VWD), a qualitative variant of VWD, is characterized by thrombocytopenia and loss of high molecular weight multimers. 2B VWF has both an increased affinity for platelet glycoprotein Ibα (GPIbα) and increased susceptibility to ADAMTS13-mediated cleavage. In order to determine whether the loss of high molecular weight (HMW) multimers associated with 2B VWD occurs due to the increased ADAMTS13 susceptibility or due to the increased binding to platelet GPIbα, we created a modified type 2B mouse model, in which the mice expressed ADAMTS13-insensitive 2B VWF. Methods: Three common type 2B VWD mutations (R1306W, V1316M, and R1341Q) were independently introduced into the mouse Vwf (mVWF) cDNA sequence containing the ADAMTS13 cleavage site knockout, Y1605A/M1606A (CSKO). Recombinant mutant mVWF was produced using transient transfection of HEK293T cells. ADAMTS13 digestion of the recombinant mVWF was performed using a Tris-Urea system. Platelet binding affinity was assessed using a botrocetin-induced platelet-binding ELISA assay. 8–9 week old C57Bl6 VWF KO mice were hydrodynamically injected with 100 μ g of naked plasmid DNA containing the liver specific ET promoter and wildtype (WT) or mutant mouse Vwf DNA. The mice were sampled weekly; complete blood counts, VWF:Ag levels, VWFpp levels, and VWF multimer structure were examined. Results: Following transient transfections, mutant and wildtype mVWF were secreted at similar levels, with a full range of VWF multimer sizes. Recombinant 2B/CSKO mVWF is markedly ADAMTS13 cleavage insensitive, with >80 U mADAMTS13 required for 50% cleavage (WT mVWF requires 1 U mADAMTS13). Similar to 2B VWF, 2B/CSKO mVWF showed enhanced platelet binding affinity in the presence of botrocetin. Hydrodynamic injections of the naked expression plasmids did not result in adverse events in the mice. Mean 2B/CSKO platelet counts at day 7 were significantly reduced compared to WT platelet counts (149 and 370, respectively). At day 14, only the mice expressing V1316M/CSKO remained thrombocytopenic, with platelet counts similar to those seen with the V1316M mutation alone (170 and 171, respectively). By day 14, the mice expressing 2B/CSKO VWF had significantly lower mean VWF:Ag levels than WT mice or mice expressing the CSKO alone (see table below). The mean VWFpp/VWF:Ag ratios were significantly increased in the 2B/CSKO mice when compared to the WT mice (1.53 and 1.00, respectively). There was no loss of HMW multimers in the mice expressing R1306W/CSKO and R1341Q/CSKO and, indeed, these mice showed significantly more multimer bands than WT mice (days 7 to 21; 14.5±0.6 and 13.9±1.2 multimer bands versus 10.2±0.5 bands for WT). In marked contrast, the mice expressing V1316M/CSKO VWF showed complete loss of high molecular weight multimers, similar to mice expressing the same 2B VWF mutation alone (8.3±1.8 and 7.7±0.9 multimer bands, respectively). Conclusions: Mice expressing R1306W/CSKO and R1341Q/CSKO VWF have an absence of thrombocytopenia and show the presence of supranormal high molecular weight VWF multimer bands. These results suggest that enhanced ADAMTS13-mediated cleavage plays an important role in the type 2B phenotype associated with these mutations. In contrast, low V1316M/CSKO VWF:Ag was associated with both thrombocytopenia and loss of HMW multimers, demonstrating that enhanced GPIbα binding affinity is the predominant mechanism associated with this mutation. Thus, the importance of ADAMTS13 susceptibility in 2B VWD is mutation-dependent, and ADAMTS13-mediated cleavage plays an important role in enhancing the severity of the type 2B VWD phenotype associated with the R1306W and R1341Q mutations. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Ultra-High Molecular Weight Polyethylene/Titanium-Hybrid Implant for Bone-Defect Replacement

Materials ◽  
2020 ◽  
Vol 13 (13) ◽  
pp. 3010 ◽  
Author(s):  
Aleksey V. Maksimkin ◽  
Fedor S. Senatov ◽  
Kirill Niaza ◽  
Tarek Dayyoub ◽  
Sergey D. Kaloshkin
Keyword(s):  
Mechanical Properties ◽  
Molecular Weight ◽  
Titanium Alloy ◽  
High Molecular Weight ◽  
Three Dimensional ◽  
Mechanical Compression ◽  
Compression Tests ◽  
Natural Bone ◽  
Ultra High Molecular Weight ◽  
Bone Layer

A hybrid implant with a structure mimicking that of natural bone was developed. Titanium alloy Ti–6Al–4V prepared with three-dimensional (3D)-printing technology was used to simulate the cortical-bone layer. The mismatch in the mechanical properties of bone and titanium alloy was solved by creating special perforations in the titanium’s surface. Porous ultra-high molecular weight polyethylene (UHMWPE) with high osteogenous properties was used to simulate the cancellous-bone tissue. A method for creating a porous UHMWPE structure inside the titanium reinforcement is proposed. The porous UHMWPE was studied with scanning electron microscope (SEM) to confirm that the pores that formed were open, interconnected, and between 50 and 850 μm in size. Mechanical-compression tests done on the obtained UHMWPE/titanium-hybrid-implant samples showed that their mechanical properties simulated those of natural bone.

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