Solubilization of porcine intestinal alpha-glucosidases and evidence for the separate identities of isomaltase and limit dextrinase.

The results of a detailed comparison of the relative efficiencies of different procedures for solubilizing the membrane-bound α-glucosidases in hog intestinal mucosa are reported. Procedures for the selective solubilization of certain members of the complex group of α-glucosidases have been developed. The selective solubilization achieved permits the conclusion that separate α-glucosidases are involved in the hydrolysis of isomaltose and the α-limit dextrins formed from starch by α-amylase.     

Encapsidation of yeast killer double-stranded ribonucleic acids: dependence of M on L.

Virus-like particles containing either L or M double-stranded ribonucleic acid (dsRNA) were isolated from a killer toxin-producing strains of Saccharomyces cerevisiae (K+ R+). At least 95% of M- and 87% of L-dsRNA were recovered in virus-like particle-containing fractions. The major capsid polypeptides (ScV-P1) of both L and M virus-like particles were shown to be identical, and 95% of the cellular ScV-P1 was found in the virus-like particle-containing fractions. Since L-dsRNA encodes ScV-P1, provision of this protein for encapsidation of M-dsRNA defines at least one functional relationship between these dsRNA genomes and associates the L-dsRNA with the killer character. If encapsidation of M-dsRNA is essential for its replication or expression, then L-dsRNA plays an essential role in maintenance or expression of the killer phenotype. The relationship between the L- and M-dsRNA genomes would be analogous to that between a helper and a defective virus. The presence of only minor quantities or uncomplexed dsRNA and ScV-P1 suggests that their production is stringently coupled.     

Low-temperature conditional cell division mutants of Escherichia coli.

Fifteen low-temperature conditional division mutants of Escherichia coli K-12 was isolated. They grew normally at 39 degrees C but formed filaments at 30 degrees C. All exhibited a coordinated burst of cell division when the filaments were shifted to the permissive temperature (39 degrees C). None of the various agents that stimulate cell division in other mutant systems (salt, sucrose, ethanol, and chloramphenicol) was very effective in restoring colony-forming ability at 25 degrees C or in stimulating cell division in broth. One of these mutants, strain JS10, was found to have an altered cell envelope as evidenced by increased sensitivity to deoxycholate and antibiotics, as well as leakage of ribonulcease I, a periplasmic enzyme. This mutant had normal rates of DNA synthesis, RNA synthesis, and phospholipid synthesis at both the nonpermissive and permissive temperatures. However, strain JS10 required new protein synthesis in the apparent absence of new RNA synthesis for division of filaments at the permissive temperature. The division of lesion in strain JS10 is cotransducible with malA, aroB, and glpD and maps within min 72 to 75 on the E. coli chromosome.     

Antioxidant activity of brown alga Saccharina bongardiana from Kamchatka (Pacific coast of Russia). A methodological approach

The present study aims to investigate the levels of polyphenols and antioxidant activity in one of the most important commercial species of seaweeds in Kamchatka, an edible brown seaweed Saccharina bongardiana. Six extracts of S. bongardiana, acetone, methanol, ethanol, and the respective 70 % aqueous solutions, were assessed for total phenol content in order to determine the most efficient extracting solvent. The total phenol content was measured by the Folin–Ciocalteu method and expressed as phloroglucinol equivalents (PGE). The antioxidant tests used were 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, linoleic acid-β carotene oxidation inhibiting assay, and Fe²⁺ ion chelating method. Higher phenolic contents were obtained using aqueous organic solvents, as compared to the respective absolute solvents; 70 % acetone was found to be the most efficient solvent (1.039 mg PGE 100 mg^?1 dry algal powder). High significant correlations were noted between total phenol content and the tested antioxidant activities; so the aqueous organic extracts exhibited the highest antioxidant activities versus DPPH radicals (EC₅₀ values of 0.6–1.1 mg dry weight (DW) mL^?1), linoleic acid-β carotene oxidation (74–78 % at 0.8 mg DW mL^?1), as well as ferrous ions (EC₅₀ values of 5.0–7.9 mg DW mL^?1). Some methodological recommendations regarding the assays used and the expression of results are proposed.     

The role of angiotensin II,endothelin-1 and transforming growth factor-β as autocrine/paracrine mediators of stretch-induced cardiomyocyte hypertrophy

Cardiac hypertrophy is a compensatory response of myocardial tissue upon increased mechanical load. Of the mechanical factors, stretch is rapidly followed by hypertrophic responses. We tried to elucidate the role of angiotensin II (AII), endothelin-1 (ET-1) and transforming growth factor- (TGF-) as autocrine/paracrine mediators of stretch-induced cardiomyocyte hypertrophy. We collected conditioned medium (CM) from stretched cardiomyocytes and from other stretched cardiac cells, such as cardiac fibroblasts, endothelial cells and vascular smooth muscle cells (VSMCs). These CMs were administered to stationary cardiomyocytes with or without an AII type 1 (AT₁) receptor antagonist (losartan), an ET-1 type A (ET_A) receptor antagonist (BQ610), or anti-TGF- antibodies. By measuring the mRNA levels of the proto-oncogene c-fos and the hypertrophy marker gene atrial natriuretic peptide (ANP), the molecular phenotype of the CM-treated stationary cardiomyocytes was characterized.Our results showed that c-fos and ANP expression in stationary cardiomyocytes was increased by AII release from cardiomyocytes that had been stretched for 60 min. Stretched cardiomyocytes, cardiac fibroblasts and endothelial cells released ET-1 which led to increased c-fos and ANP expression in stationary cardiomyocytes. ET-1 released by stretched VSMCs, and TGF- released by stretched cardiac fibroblasts and endothelial cells, appeared to be paracrine mediators of ANP expression in stationary cardiomyocytes.These results indicate that AII, ET-1 and TGF- (released by cardiac and vascular cell types) act as autocrine/paracrine mediators of stretch-induced cardiomyocyte hypertrophy. Therefore, it is likely that in stretched myocardium the cardiomyocytes, cardiac fibroblasts, endothelial cells and VSMCs take part in intercellular interactions contributing to cardiomyocyte hypertrophy.