Effector-assisted refolding of recombinant tissue-plasminogen activator produced in escherichia coli

Recombinant tissue-plasminogen activator (r-tPA), expressed in Escherichia coli cells in an aggregated form, was solubilized with a strong chaotrope in the absence of any reducing agent. The solubilized molecule was reactivated by a procedure that was developed to mimic the physiological conditions optimal for the functional folding and activity of the native protein. The use of partially purified fibrinogen, as a source of fibrin (the effector), is shown to facilitate the reactivation process and increase its yield by at least a factor of two. The yield of the process is also shown to be particularly dependent on the recombinant protein concentration. At a concentration level of 3-3.7 mg r-tPA/L in the reactivation mixture, up to a 90% yield of activity was obtained. Purification of the activated form of r-tPA was achieved with a two-step column-chromatography scheme. This included a gel filtration step on a Sephadex G-50 column followed by an affinity chromatography step on a lysine-sepharose column. The product was composed of roughly equal amounts of one-chain and two-chain t-PA. The feasibility of using a two water-soluble polymeric phase system, with a centrifugal partition chromatography (CPC), in scaling up the reactivation process or the purification step was also evaluated.     

Synthesis and characterization of polyaniline nanoparticles in the presence of magnetic field and samarium chloride

A new and economical method of preparing polyaniline (PANI) nanoparticles will be introduced in this article. Compared with conventional methods, this method is much more simple and convenient. Scanning electron microscope (SEM) shows that the diameter of particles are between 30 and 50 nm, which is in good agreement with the results of a transmission electron microscope (TEM). Polyaniline/SmCl₃/Bp, polyaniline/SmCl₃ and polyaniline/HCl were prepared in a solution containing 1.0 mol dm⁻³ aniline, 1.0 mol dm⁻³ HCl with and without 0.5 mol dm⁻³ SmCl₃, in the presence and in the absence of a magnetic field, respectively. Their conductivity, UV-vis spectra, FTIR spectra, X-ray diffraction (XRD) and thermogravimetric analysis (TGA) were investigated. Changes in UV-vis and FTIR spectra indicate a strong interaction between Samarium ions (SmCl₃) and polyaniline chains. The conductivity of PANI depends on magnetization and concentration of Sm³⁺. Polyaniline/SmCl₃/Bp has the higher degree of crystallinity than that of polyaniline/HCl.     

Solvation of iron and chromium complexes in alcohol–water mixtures

Solubilities are reported for the perchlorates of five iron(II)-diimine complexes in t-BuOH–H₂O and one in MeOH–H₂O mixtures, for three iron(III)-3-hydroxy-4-pyranonate and three iron(III)-3-hydroxy-4-pyridinonate complexes in MeOH–H₂O and t-BuOH–H₂O, and for two chromium(III)-3-hydroxy-4-pyranonate complexes in MeOH–H₂O. Transfer chemical potentials are thence derived for the various iron(II), iron(III) and chromium(III) complexes, for transfer from H₂O into the respective mixed solvents (at 298.2 K). These results are combined with values reported earlier for related complexes, and for other alcohol–H₂O mixtures, to give an overall picture of solvation, expressed in the thermodynamic format of transfer chemical potentials, for iron(II)-diimine, iron(III)-3-hydroxy-4-pyridinonate and chromium(III)-3-hydroxy-4-pyranonate complexes in H₂O-rich aqueous-alcohol mixtures. Some spectroscopic (¹H-n.m.r.; i.r.) and kinetic (aquation rate constants at 298.2 K) data are reported for the chromium(III) complexes.     

Properties of glass-fiber reinforced composites based on liquid diglycidyl ether of bisphenol-c-epoxy resin

Glass-fiber reinforced epoxy composites were fabricated from the matrix resin liquid diglycidyl ether of bisphenol-C (DGEBC) using various amines as curing agents with and without fortifier (20 phr). The epoxy laminates were evaluated for their mechanical properties, such as flexural strength, interlaminar shear strength (ILSS), tensile strength and shore-D hardness. Dielectric properties, such as the dielectric constant, tan δ, dielectric loss and the resistivity of the laminated samples, were measured. The effect of the chemical reagents on the mechanical properties (i.e. flexural strength, lLSS) was also studied.     

Characterization of Fatty Acids,Polysaccharides, Amino Acids,and Minerals in Marine Macroalga Chaetomorpha crassa and Evaluation of Their Potentials in Skin Cosmetics

Cosmetic industries are highly committed to finding natural sources of functional active constituents preferable to safer materials to meet consumers’ demands. Marine macroalgae have diversified bioactive constituents and possess potential benefits in beauty care products. Hence, the present study was carried out to characterize the biochemical profile of marine macroalga Chaetomorpha crassa by using different techniques for revealing its cosmetic potentials. In results, the FTIR study characterized the presence of different bioactive functional groups that are responsible for many skin-beneficial compounds whereas six and fifteen different important phycocompounds were found in GCMS analysis of ethanolic and methanolic extracts, respectively. In the saccharide profile of C. crassa, a total of eight different carbohydrate derivatives were determined by the HRLCMS Q-TOF technique, which showed wide varieties of cosmetic interest. In ICP AES analysis, Si was found to be highest whereas Cu was found to be lowest among other elements. A total of twenty-one amino acids were measured by the HRLCMS-QTOF technique, which revealed the highest amount of the amino acid, Aspartic acid (1207.45 nmol/mL) and tyrosine (106.77 nmol/mL) was found to be the lowest in amount among other amino acids. Their cosmetic potentials have been studied based on previous research studies. The incorporation of seaweed-based bioactive components in cosmetics has been extensively growing due to its skin health-promoting effects.     

Al–SnSe2 Schottky Barrier

Shottky barrier diodes are fabricated on SnSe₂ crystal grown by direct vapour transport method. The barrier is produced by thermal evaporation of aluminium on as grown single crystal of SnSe₂. The barrier height of Schottky diodes are estimated using current-voltage characteristics, and activation energy measurement. The barrier height is estimated about 0.49 eV. The implications are discussed.     

Fabrication and characterization of ZnTe/CdSe thin film solar cells

Thin film solar cells have been prepared and investigated by using indium-doped n-CdSe in conjunction with p-ZnTe. The films are prepared on indium tin oxide (ITO) coated glass substrates by the thermal evaporation. Thin film layers have been deposited without breaking vacuum by utilizing a mask rotating system. Silver has been used as a stimulator for bringing improvement in the crystallinity and electrical behaviour of ZnTe thin films. Indium is used for obtaining the back ohmic contacts. The solar cells thus produced have been characterized by I–V-, C–V-, and spectral response measurements. The maximum open circuit voltage, the short circuit current density, and the efficiency are obtained as 415 mV, 11.60 mA cm⁻² and 1.86%, respectively under 100 mW cm⁻² input optical power. The diffusion potential and depletion width are found to be 1.42 eV and 21 nm, respectively.     

Analysis of cross-linked human hemoglobin by conventional isoelectric focusing, immobilized pH gradients, capillary electrophoresis, and mass spectrometry.

Diaspirin cross-linked hemoglobin (DCLHb), a hemoglobin-based oxygen carrier exhibiting near physiological oxygen binding capability and devoid of nephrotoxic side effects, was previously found, by gel permeation, reversed-phase high performance liquid chromatography (RP-HPLC) and mass spectrometry, to consist of ca. 94% cross-linked product (reacted on the Lys 99 of two alpha-chains), accompanied by ca. 6% cross-linked Hb, which also reacted on the Lys 132 and/or Lys-144 of the beta-chains and a small amount of intermolecularly cross-linked dimers. However, conventional isoelectric focusing in carrier ampholyte buffers (CA-IEF) gave an unexpected spectrum of four major, almost equally represented, pI species in the pH range of 6.82-7.01, a band of mid-intensity with a pI of 7.11, and two minor components with pls of 6.73 and 6.77. This extraordinary polydispersity was reevaluated by other surface charge probes, such as immobilized pH gradients (IPG) and capillary zone electrophoresis (CZE) of native and denatured globin chains. IPGs of DCLHb gave the expected spectrum of bands, consisting of a main component (92%) with pl 7.337 and three additional minor bands, with lower pIs, representing ca. 8% of the total. These data were in agreement with CZE profiles of native DCLHb, which resolved, in addition to the main DCLHb peak, 3-4 minor components representing ca. 10% of the total. Also, CZE of denatured, heme-free globin chains gave the expected pattern with only traces of minor, extrareacted species. The latter technique, in addition to resolving alpha- and beta-globin chains in a 1:1 ratio in control Hb, resolved a free beta- and the alpha-alpha-dimer in DCLHb. In a 1:1 mixture of control and DCLHb, three peaks were observed, eluting in the order alpha-, alpha-alpha- and beta-globin chains. The identity of the major DCLHb and of the minor species was ascertained by mass spectrometry.     

Ethyl 3‐amino‐6‐phenyl‐4‐tolylthieno[2,3‐b]pyridine‐2‐carboxylate

In the title compound, C₂₃H₂₀N₂O₂S, the central thieno­pyridine ring system is essentially planar, the dihedral angle between the planes of the two rings being 0.3 (2)°. The terminal ethyl carboxyl­ate group is twisted by 26.7 (3)° away from the central ring system. A short intramolecular hydrogen bond involving the amino N atom and the carbonyl O atom [N⋯O = 2.806 (4) Å] forms a pseudo‐six‐membered ring. Significant intermolecular C—H⋯N, C—H⋯O and C—H⋯π interactions contribute strongly to the stability of the structure, along with weak π–π‐stacking interactions.