Ultracentrifugal Study of Liposome Solubilization by Sodium Dodecylsulfate

2002 ◽  
Vol 23 (4) ◽  
pp. 483-490 ◽  
Author(s):  
Hyungki Lee ◽  
Namita Deo ◽  
P. Somasundaran
1987 ◽  
Author(s):  
L Goretzki ◽  
E Miller ◽  
A Henschen

Plasmin and leucocyte elastase are regarded as the two medically most important fibrin(ogen)-degrading proteolytic enzymes. There is, however, a considerable difference in information available about the cleavage specificities and fragmentation pathways of these two enzymes. Degradation by plasmin has been studied already for a long time in great detail so that now the time course of the degradation, the cleavage sites and the functional properties of many fragments are well known. In contrast, relatively little is known about the degradation by leucocyte elastase, except that the overall cleavage pattern resembles that obtained with plasminIn this investigation the leucocyte elastase-mediated degradation of fibrinogen has been examined by means of proteinchemi-cal methods. Human fibrinogen was incubated with human enzyme material for various periods of time and at some different enzyme concentrations. The split products formed at the various stages were isolated in pure form by gel filtration followed by reversed-phase high-performance liquid chromatography. The fragments were identified by N-terminal amino acid sequence and amino acid composition. The course of the degradation was also monitored by sodium dodecylsulfate-polyacrylamide gel electrophoresis. All cleavage patterns were compared with the corresponding patterns from plasmic degradation. It could be confirmed that X-, D- and E-like fragments are formed also with elastase. However, several early elastolytic Aα-chain fragments are characteristically different from plasmic fragments. The previously identified N-terminal cleavage site in the Aα-chain, i.e. after position 21, was found to be the most important site in this region of fibrinogen. The very early degradation of the Aα-chain N-terminus by elastase is in strong contrast to the stability against plasmin. Several cleavage sites in N-terminal region of the Bβ-chain were observed, though the low amino acid specificity of elastase partly hampered the identification. The γ-chain N-terminus was found to be as highly stable towards elastase as towards plasmin. The results are expected to contribute to the understanding of the role of leucocyte elastase in pathophysiologic fibrino(geno)lysis


2014 ◽  
Vol 79 (4) ◽  
pp. 457-468 ◽  
Author(s):  
Jaroslav Katona ◽  
Sandra Njaradi ◽  
Verica Sovilj ◽  
Lidija Petrovic ◽  
Brankica Marceta ◽  
...  

Rheological properties of mixtures of hydroxypropylmethyl cellulose (HPMC), a nonionic associative cellulose ether, and sodium dodecylsulfate (SDS), an anionic surfactant, were investigated by viscosity measurements performed at different shear rates (0.1-6000 s-1). HPMC/SDS mixtures containing different concentrations of SDS (CSDS=0.00-3.50 % w/w) and HPMC concentrations which corresponded to the overlap parameter c/c*=3, 6, and 12 were prepared. All HPMC/SDS mixtures were found to be shear-thinning when examined in a low-end-to mid-range of the applied shear rates. The degree of shear-thinning, n, and viscosity of the mixtures were influenced by composition of HPMC/SDS mixtures and HPMC-SDS complex formation. The changes in n ranged from values typical for highly shear thinning to almost perfectly Newtonian liquids, and were more pronounced as c/c* was increased from 3 to 6 and 12. A change in flow profile and a buildup of the first normal stress difference (N1) was observed in HPMC/SDS mixtures with c/c*=6 and 12 and CSDS 0.55-1.00 % and 0.55-2.50 %, respectively, when a critical shear rate, crit. was exceeded, suggesting that a shear-induced structure formation in the mixtures took place.


2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Francesco Amadeo ◽  
Gianluca Polvani ◽  
Marco Agrifoglio ◽  
Federica Boschetti ◽  
Maurizio Pesce ◽  
...  

Background: Aldehyde-fixed pericardium is commonly used in valve implant manufacturing. Despite its wide employment, this tissue undergoes chronic rejection that limits implant performance and durability. In this work, we employed a method to engineer a leaflet-like tissue by seeding aortic valve interstitial cells (VIC) into fixative-free decellularized pericardium using a bioreactor based approach. Methods: Following treatment with hypotonic buffer (Tris-HCl) to induce cell lysis, porcine pericardium was incubated with TritonX-100, to remove adipose tissue and then treated with sodium dodecylsulfate to wash cellular debris. DNA was removed by incubation into a DNAse I solution. Pericardium permeability was measured on samples before and after decellularization (pressure from 735 Pa to 2200 Pa). A direct perfusion bioreactor was employed to seed (3days, 3ml/min) porcine VICs (6.5E+5 cell/scaffold) into decellularized pericardium patches (6mm diameters) and perform long-term culture (up to 14 days, 0.03ml/min). Cell seeding efficiency (Day 3) and cell proliferation (Day 7-14) were evaluated by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) staining, histological analyses and fluorescence staining for quiescent/activated VIC markers (DAPI, vimentin, α-smooth muscle actin). Results: Permeability tests revealed a significant increase in decellularized samples (1-way ANOVA p < 0.05), thus supporting its use in a direct perfusion bioreactor system. MTT staining revealed homogeneous cell seeding distribution, supported by DAPI staining, showing efficient cellularization through the whole patch volume. Computer-based cell nuclei counting revealed a significant cell increase from day 3 to 7 and 14 ( p <0.05 1-way ANOVA). Immunofluorescence showed a marked reduction of αSMA in cells populating the inner layers. Conclusions: Our data show, for the first time, the capability to seed and culture VICs into a cell/fixative free pericardium with a direct perfusion system. Moreover, the use of pericardium treated with our decellularization procedure and recellularized under dynamic conditions supports a more physiological growth of VIC, as suggested by downregulation of αSMA in the inner pericardium layer.


2019 ◽  
Vol 6 (3) ◽  
pp. 181979 ◽  
Author(s):  
K. M. Sachin ◽  
Sameer A. Karpe ◽  
Man Singh ◽  
Ajaya Bhattarai

The micellar property of mixed surfactant systems, cationic (dodecyltrimethylammonium bromide, DTAB) and anionic (sodium dodecylsulfate, SDS) surfactants with variable molar ratios in aqueous system has been reported by using surface tension and conductivity measurements at T = 293.15, 298.15 and 303.15 K. DTAB concentrations are varied from 1.0 × 10 −4 to 3 × 10 −4 mol l −1 in 1.0 × 10 −2 mol l −1 SDS solution while the SDS concentration is varied from 1.0 × 10 −3 to 1.5 × 10 −2 mol l −1 in approximately 5.0 × 10 −3 mol l −1 DTAB, so that such concentrations of DTAB-SDS (DTAB-rich) and SDS-DTAB (SDS-rich) solutions were chosen 3 : 1 ratio. The critical micellar concentration, as well as surface and thermodynamic properties for DTAB-rich and SDS-rich solutions, were evaluated by the surface tension ( γ ) and conductivity ( κ ) methods. The pseudo phase separation model was coupled with the dissociated Margules model for synergism. The Krafft temperature behaviour and optical analysis of mixed surfactants are studied using conductivity and UV–Vis spectroscopy, respectively. The dispersibility and stability of DTAB-rich and SDS-rich solutions with and without dyes (2.5 × 10 −5 mol l −1 of methyl orange and methylene blue) are carried out by using UV–Vis spectroscopy and dynamic light scattering.


Sign in / Sign up

Export Citation Format

Share Document