Interaction of apolipoprotein B-containing lipoprotein secreted by Hep G2 cells with receptors for low-density lipoprotein.

Radioligand and immunoenzymatic techniques were used to characterize the receptor binding properties of apolipoprotein B-containing lipoprotein produced by HepG2 cell line (H-LpB). It was found that compared to plasma low-density lipoprotein (LDL), the interaction of H-LpB nonseparated from conditioned medium with normal fibroblasts was 6-8-times lower and only slightly exceeded the nonspecific binding of LDL modified by malondialdehyde, while the uptake of the indicated lipoproteins by LDL receptor-negative strain of fibroblasts were identical. The uptake of H-LpB by normal fibroblasts increased 1.5-2-times after isolation from the culture medium by immunoaffinity chromatography. The effect of isolation could be explained by the finding that apolipoprotein E-containing lipoprotein secreted by HepG2 cells effectively competed for the binding with LDL-receptors. The obtained results suggest that H-LpB produced by HepG2 cells is poorly recognized by the LDL-receptors.     

Electrostatic effects in the alpha-chymotrypsin-catalyzed acyl transfer. II. Efficiency of nucleophiles bearing charged groups in various locations

We investigated the alpha-chymotrypsin-catalyzed acyl transfer to a series of glycine oligomers. It could be established that the electrostatic interactions between the carboxylate group of the nucleophiles and the S'-subsites of the enzyme fall off with the length of the nucleophile molecule. Additional negatively charged residues in the nucleophile lead to a considerable reduction of the acyl transfer efficiency. An arginine residue in P'1- or P'3-position, but not in P'2-position, makes favourable interactions with the appropriate S'-subsites of the enzyme.     

Change in charge of the erythrocyte membrane during exposure to a pulsed electromagnetic field]

By means of hydrophobic anion phenyldicarbaundecaboron electrophoresis in PAA gels and filtration through "Millipore" filters an increase of the total membrane charge and decrease in the amount of membrane-bound proteins (actin, glyceraldehyde-3-phosphatedehydrogenase) after erythrocyte ghosts exposition in pulsed electromagnetic field (PEMF, 1-10 Hz pulse frequency, 24 mTesla) were demonstrated. The effects of PEMF are explained by the changes in the distribution of electrostatic charges near the membrane protein-protein binding sites.     

The role of 4-oxyproline in the phylogenetic change of collagen thermostability]

Analysing the data on imino acid content and thermostability of collagens distinguished phylogenetically it is shown that one hydroxyproline residue (per 100 residues) which is localized in the third position of the triplet increases thermostability per 0.6 degree in the average. The magnitude obtained exceeds the value 4 times (0.16 degree). This happens at the expense of hydroxylated proline in the third position.     

Plasma sex steroid-binding protein in mature heifers: effects of reproductive status, nutritional level, and porcine growth hormone, and estradiol-17 beta treatments

In heifers, the specific 5 alpha-dihydrotestosterone (5 alpha-DHT) binding to sex steroid-binding protein (SBP) was 53 nM in midluteal phase and was 20% lower (p less than 0.05) in early luteal phase. In ovariectomized heifers on a high (H) or a low (L) energy diet, SBP bound testosterone with high specificity and estradiol-17 beta (E2) with a lower specificity; the affinity constant with 5 alpha-DHT was similar in the two groups independent of diet (Ka = 1.0 and 1.3 x 10(9) x M-1 for H and L heifers, respectively). Electrophoretic mobility of SBP was not affected by undernutrition. The specific 5 alpha-DHT binding to SBP in anestrous heifers compared to heifers in early luteal phase was 17% lower 18 days after cessation of ovarian activity (p less than 0.05) and 35% lower after 78 days of anestrus (p less than 0.001). Specific 5 alpha-DHT binding to SBP was 33% lower (p less than 0.001) in L heifers than in H heifers (40 nM). Porcine growth hormone (pGH) and E2 injections increased specific 5 alpha-DHT binding to SBP in both H and L heifers, but the effect depended on diet. Simultaneous injections of pGH and E2 increased specific binding in H heifers (p less than 0.02), whereas it had no effect in L heifers. We conclude that SBP binding capacity varies with ovarian activity in heifers and that nutritional status is one factor of regulation of SBP binding capacity because it affects binding and modifies the SBP response to stimulating factors such as pGH and E2.     

DNA interpolyelectrolyte complexes as a tool for efficient cell transformation.

A tool was developed for enhancement of plasmid penetration into an intact cell, based on increasing DNA hydrophobicity via inclusion into a soluble interpolyelectrolyte complex (IPC) with polycations. The characteristics of formation of DNA IPC with synthetic polycations [poly(N-ethyl-4-vinylpyridinium)bromide (PVP) and PVP modified with 3% of N-cetyl-4-vinylpyridinium units (PVP-C)] were studied using ultracentrifugation and polyacrylamide gel electrophoresis methods. The conditions were established under which the mixing of DNA and polycation aqueous solutions results in the self-assembly of soluble IPC species. Incorporation of DNA into IPC results in the enhancement of DNA binding with isolated Bacillus subtilis membranes. A considerable increase in the efficiency of transformation of B. subtilis cells with pBC16 plasmid resulted from incorporation of the plasmid into the IPC with PVP and CVP.     

A novel magnetic silica support for use in chromatographic and enzymatic bioprocessing

A limited number of support matrices have so far been developed for use in magnetically stabilized fluidized bed (MSFB) applications. We have developed a versatile magnetic silica support which can be derivatized readily for both adsorption chromatography and enzyme immobilization by well-known techniques. A magnetic pellicular bead is prepared by electrostatically depositing alternating layers of colloidal silica and cationic polymer onto macroscopic nickel core particles. The polymer is then burned out and the silica partially sintered to yield a porous shell with 5-80 m(2)/g of surface area. This magnetic composite was tested as a support for immobilizing invertase. Sucrose was continuously converted to its component monosaccharides with nearly constant activity over the first 8 days and retention of 50% of initial activity after 25 days.     

Active immunization to exogenous and endogenous antidepressants. The formation of antibodies to biogenic amines and peptide regulators

The active immunization of rabbits and white rats to antidepressant sydnophen results in the formation of antibodies binding norepinephrine, dophamine, serotonine as well peptide regulatory compounds: substance P, pynorphine, vasopressin and beta-endorphin. The immunization against endogenic antidepressant thyroliberin induces the formation of antibodies to the same biogenic amines and to the gamma-aminobutyric acid, oxytocin and leu encephalin. The data obtained are discussed in connection with some physiological and biochemical changes found earlier during immunization to antidepressants.     

The effect of the acidity of the medium on the magnitude of the light-dependent antibacterial activity of chlorophyllides]

The influence of medium acidity on the intensity of light-dependent antibacterial activity of chlorophyllides isolated from biomass of microscopic green alga Westella botryoides has been studied. This light-dependent antibacterial activity has been shown to be maximum at low pH values. The ability to suppress the bacterial growth under the action of light decreases in neutral and alkaline media. It has been concluded that the reason of increase of light-dependent antibacterial activity of chlorophyllides at low pH values is formation of pheophorbides out of them     

100ps molecular dynamic simulation of d(TATCACC)2.

A heptanucleotide sequence d(TATCACC)2 from OR3 region of bacteriophage lambda is considered sufficient for the recognition of Cro protein. We present here results on molecular dynamic simulations on this sequence for 100 ps in 0.02 ps interval. The simulations are done using computer program GROMOS. The conformational results are averaged over each ps. The IUPAC torsional parameters for 100 conformations are illustrated using a wheal and a dial systems. Several other stereochemical parameters such as H-bonding lengths and angles, sugar puckers, helix twist and roll angles as also distances between opposite strand phosphorus are depicted graphically. We find that there is rupture of terminal H-bonds. The bases are tilted and shifted away from the helix axis giving rise to bifurcated H-bonds. H-bonds are seen even in between different base pairs. The role of these dynamic structural changes in the recognition of OR3 operator by Cro protein is discussed in the paper.