A new method for rapeseed hulls purification – Proof of concept

A new method for the purification of Qβ RNA-dependent RNA polymerase

In this paper a new method of purification of raw mixtures of alkaloids by dialysis of aqueous solutions of their salts through a semipermeable membrane is described. The results obtained by using this method of purification of raw mixtures of alkaloids from plant extracts and mother liquors after crystallization of the major alkaloid have been given. In all cases under investigation this method of purification gave crystalline substances.

Nearly 3 decades ago, a dendritic structure was stepwise synthesized for the first time as a new type of molecules with promising applications. During years a huge effort has been devoted to implement the synthetic skills concerning the synthesis of these molecules and especially, new methods for purification and characterization of these compounds that are in the nanoscale range. The chemical manipulation of the surface and inner core of dendrimers were strategically used to allow a tailor-made control of physical-chemical properties and to discover new applications in material science and biomedicine. Although several examples have been reported in the literature describing applications of functionalized dendrimers and acclaiming a key role of these molecules, very scarce examples are actually close to the market. This review summarizes the state of the art of dendrimers and dendritic polymers as anti-infective agents, with a special focus on the strategies to block receptors used by pathogens for attachment, cell entry and dissemination. These nanometre size molecules are very attractive compounds as new drugs easily to be manipulated to improve their activity and scope. This is already a very active area of research, where we are involved, with interesting potential as demonstrated by the Phase I clinical trial of a functionalized dendrimer with real possibilities to reach the market soon. The success of this compound should provoke an enormous stimulus to scientists working in this area as well as in the industrial companies for investment in this topic. Keywords: pathogen-associated molecular patterns, pattern recognition receptors, C type lectin receptors, Dendritic-Cell-Specific ICAM-3 Grabbing Non integrin, Cholera Toxin, Vero Toxins, Streptococcus suis

The purification and characterization of Dictyostelium discoideum plasma membranes

Summary Active C′1s was isolated from the euglobulin fraction of human serum by column chromatography on CM and DEAE cellulose and Pevikon block electrophoresis. The final product revealed a single precipitin line against horse anti-whole human serum on immunoelectrophoresis and did not contain C′1q. A spectrophotometric assay was introduced for measuring the esterase activity of C′1s using TAMe as a substrate. Active C′1s was capable of generating SAC′4,2a although it did not bind to EAC′4 cells and EA. The esterase activity and the hemolytic activity as well as the C′2 and C′4 destructive activity of active C′1s were associated with a single homogeneous protein which migrated to the α-globulin region and had a molecular weight of approximately 110,000. A molecular titration of active C′1s is not possible because active C′1s transfers from site to site; however, SAC′4,2a generating activity could be correlated with TAMe esterase activity. As the ratio of hemolytic to esterase activity of active C′1s was 20 to 40 times less than that of C′1a, the firm binding of C′1a mediated through the combining site for antibody, presumably on the C′1q subunit, increases the efficiency of cleavage and transfer of C′2 at SAC′4.

Authentic hSAA related with AA amyloidosis: New method of purification, folding and amyloid polymorphism

A New Method of Purification and Sensitive Bioassay of Platelet-Activating Factor (PAF) in Human whole Blood.

A procedure of isolation of oxalate oxidase from barley seedling and a new method for purification is described. A purification step was accomplished by the second partition using polyethylene glycol (PEG-6000) and Devi-Iran-500 (4.5%1 4), and affinity chromatography using oxalate immobilized on activated CNBR-Sepharose 6MB as a ligand. After partition some protein were separated, and the specific activity were increased by 5-30 fold. The affinity chromatography using Oxalate-Sepharose effectively separated the oxalate oxidase. The specific activity of pure enzyme was 154.3 U/mg protein and the purity was 41 fold.

Recovery of antithrombin III from milk by expanded bed chromatography

A new affinity gel for purification of carbonic anhydrase isozymes was prepared using EUPERGIT C-250L derivatized with p-aminobenzenesulfonamide, an inhibitor of carbonic anhydrase. The binding capacity of the affinity gel was determined at different temperatures, pH values, elution buffers, and ionic strengths. Human carbonic anhydrase isozymes (HCA I and HCA II) and bovine carbonic anhydrase (BCA) were purified in high yields from erythrocytes.

A new method for purification of mature elastin

Enhancement of Smooth Muscle Contraction with Protein Phosphatase Inhibitor 1: Activation of Inhibitor 1 by cGMP-Dependent Protein Kinase

High specific activity insulin-I¹³¹ can be prepared with oxidizing agents, but these methods are associated with chemical damage of the insulin. This can be overcome by a suitable purification technique. A new method of purification of insulin-I¹³¹ was developed in which the damaged iodinated insulin is removed by buffer elution on a dextran column. In this way, using chloramine T as an oxidizing agent, and dextran column purification, it was possible to prepare insulin-I¹³¹ with a specific activity of 1 c/mg, the final product containing less than 3% damaged insulin. Crystalline bovine insulin (2.5 mu g in 0.025 ml pH 8 borate buffer) and 4.5 mc I¹³¹ in 0.05 ml sodium hydroxide were mixed with 50 mu g chloramine T in 0.025 ml borate buffer for 1 min. The reaction was terminated by adding 120 mu g sodium metabisulfite in 0.1 ml buffer. Free I¹³¹ was removed by dialyzing the iodinated-insulin mixture against distilled water. The iodinated insulin solution was passed through a 1-g Sephadex G-75 column made up with barbitone buffer pH 8.5. The damaged insulin-I/sup 131/ came out in the first 3-ml elute fraction. Purified insulin-I¹³¹ was collected in later fractions. With chloramine T oxidative system, an iodinationmore » efficiency over 50% was achieved. (H.H.D.)« less

Review of Astragalus membranaceus polysaccharides: Extraction process, structural features, bioactivities and applications

A new method for purification of “polypeptide synthetase” from Alcaligenes faecalis is described in which nucleic acids are removed by purification on DEAE‐cellulose with a recovery of 60–75% of the initial enzymatic activity.The enzyme preparation catalyses the binding of the naturally occuring l‐amino acids to a messenger‐like RNA whose properties have been previously described. The reaction requires the addition of any one of the four nucleoside triphosphates (ATP, GTP, CTP or UTP), which is degraded to the corresponding nucleoside diphosphate and orthophosphate. For certain amino acids, however, the requirement for a nucleoside triphosphate is limited to two of the four.The purified enzyme preparation, unlike crude extracts, does not catalyse an amino dependent ATP‐[32P]PPi exchange. Neither crude extracts, nor purified preparations show detectable exchange between [32P]PPi and GTP, CTP, or UTP in the presence or in the absence of l‐amino acids.The presence of more than one enzyme in the purified preparation was demonstrated by further substantial fractionation of [14C]leucine binding activity and separation of this from [14C]alanine binding activity. However, no fractionation, which was dependent on a specific nucleoside triphosphate, could be achieved.