Diazepam induces relaxation of chick embryo muscle fibers in vitro and inhibits myosin synthesis

Diazepam (Valium/Roche) causes an immediate cessation of spontaneous contraction in chick embryo skeletal muscle fibers growing in vitro. Between 24–48 h later in the presence of 100 μM diazepam the relaxed muscle fibers no longer accumulate myosin as measured by the total amount of myosin heavy-chain peptide extracted from the cell cultures and identified by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The myosin heavy chain assay procedure was standardized by quantitative precipitation of myosin with antibody to column purified chicken skeletal muscle myosin. Failure to accumulate myosin is related to a progressive inhibition of myosin synthesis. Diazepam-treated cultures showed an 80% inhibition of myosin heavy-chain synthesis over a period of 4 days. At the same time the rate of myosin heavy-chain degradation increases in diazepam-treated cultures relative to matched control cultures. Total protein synthesis was only marginally affected suggesting that diazepam may differentially inhibit myofibrillar protein synthesis. All of the observed effects of diazepam were reversible if drug exposure was limited to 48 h. The apparent specificity and reversibility of diazepam suggests that the drug will be useful in probing the mechanisms of terminal skeletal muscle cell differentiation and the hypotrophic relationship between chronic relaxation and inhibition of accumulation of myosin and perhaps other myofibrillar proteins.     

Liver as a source of transformed cell growth factor

Isolated rat hepatocytes release an acidic glycoprotein(s) that can selectively promote the growth of transformed cells. This factor has a molecular weight of 60 000–70 000 D. Liver microsomal and cytosol fractions contain two species of stimulatory activity—44 000 and 3 500 D. Mitochondrial and nuclear fractions contain only the lower molecular weight factor.     

The Prophage of SPβc2DcitK1, a Defective Specialized Transducing Phage of BACILLUS SUBTILIS

The defective specialized transducing phage SP beta c2dcitK1 carries two known bacterial genes, kauA and citK, as well as SP beta hage markers including the heat-sensitive repressor allele, c2. Some phage genes (including essential ones) are missing. When SP beta c2dcitK1 transduces SP beta-sensitive cells of Bacillus subtilis, the defective prophage is inserted into sites in the homologous bacterial DNA of the attSP beta-kauA-citK region of the recipient chromosome. During the growth of these transductants, occasional excisions occur that result in the loss of the phage genes and of the heterogenotic state. These excisions increase greatly in frequency during growth at repressor-inactivating temperatures. The kinds of insertions and excisions seen suggest that a Campbell-type (CAMPBELL 1962) circular phage genome may occur transiently. If the transductants are superinfected by SP beta c2 or by the clear-plaque mutant SP beta c1, the resulting double lysogen can be heat induced to release high-frequency-of-transduction (HFT) lysates for kauA and citK.     

Congenital Dislocation of the Hip: Prognostic Implications of Early Diagnosis

A retrospective analysis of 62 patients with 78 congenital dislocations of the hip (CDH) referred to Children''s Hospital at Stanford was done to ascertain whether CDH was being diagnosed as early as had been thought possible and whether various treatments were effective. In this series there were a substantial number of cases of CDH that were not diagnosed within the first 48 hours of life. At Children''s Hospital the Pavlik harness was the most effective form of treatment of congenital dislocations of the hip in the perinatal period.     

Extracellular Maltase of Bacillus brevis

Bacillus brevis NRRL B-4389 produced extracellular maltase (alpha-glucosidase; EC 3.2.1.20) only in the presence of short alpha-1,4-glucosidic polymers, such as maltose and maltotriose. An optimum medium was developed; it contained 2.5% maltose, 0.5% nonfat dry milk, 0.4% yeast extract, and 0.01% CaCl(2). The enzyme was produced extracellularly during the logarithmic phase of growth; no cell-bound activity was detected at any time. Partial purification of the maltase was accomplished by using diethylaminoethyl cellulose batch adsorption, ammonium sulfate precipitation, and Sephadex G-200 gel filtration. Maltase, isomaltase (oligo-1,6-glucosidase), and glucosyltransferase activities were purified 20.0-, 19.1-, and 11.5-fold, respectively. Some properties of the partially purified maltase were determined: optimum pH, 6.5; optimum temperature, 48 to 50 degrees C; pH stability range, 5.0 to 7.0; temperature stability range, 0 to 50 degrees C; isoelectric point, pH 5.2; and molecular weight, 52,000. The relative rates of hydrolysis of maltose (G(2)), maltotriose (G(3)), G(4), methyl-alpha-d-maltoside, G(40), dextrin, and isomaltose were 100, 22, 12, 10, 10, 8, and 5%, respectively; the K(m) on maltose was 5.8 mM; d-glucose, p-nitrophenyl-alpha-d-glucoside, and tris (hydroxymethyl) aminomethane were competitive inhibitors; transglucosylase activity of the enzyme on maltose resulted in the synthesis of isomaltose, isomaltotroise, and larger oligosaccharides.     

Preparation and crystal structure of [NEt4]3[Fe6W2S8(SEt)9]; structural and electrochemical comparisons with its molybdenum analogue

[NEt₄]₃[Fe₆M₂S₈(SEt)₉] (M = Mo or W) compounds are isomorphous and contain molybdenum and tungsten atoms in an essentially identical environment. These complexes undergo an irreversible one-electron oxidation at −0.46 V (Mo) and −0.51 V (W) and two one-electron reductions at −1.56 and −1.76 V (Mo) and −1.52 and −1.84 V (W), in DMSO solution versus (0.1 M). The only distinction between the behavior of these molybdenum and tungsten complexes identified thus far is that, for the former the reductions are reversible whereas for the latter they are irreversible. This difference may be relevant to the low activity found for nitrogenases reconstituted with tungsten in place of molybdenum.     

Does high-mobility-group non-histone protein HMG 1 interact specifically with histone H1 subfractions?

The interaction of the non-histone chromosomal protein HMG (high-mobility group) 1 with histone H1 subfractions was investigated by equilibrium sedimentation and n.m.r. sectroscopy. In contrast with a previous report [Smerdon & Isenberg (1976) Biochemistry 15, 4242--4247], it was found, by using equilibrium-sedimentation analysis, that protein HMG 1 binds to all three histone H1 subfractions CTL1, CTL2, and CTL3, arguing against there being a specific interaction between protein HMG 1 and only two of the subfractions, CTL1 and CTL2. Raising the ionic strength of the solutions prevents binding of protein HMG 1 to total histone H1 and the three subfractions, suggesting that the binding in vitro is simply a non-specific ionic interaction between acidic regions of the non-histone protein and the basic regions of the histone. Protein HMG 1 binds to histone H5 also, supporting this view. The above conclusions are supported by n.m.r. studies of protein HMG 1/histone H1 subfraction mixtures. When the two proteins were mixed, there was little perturbation of the n.m.r. spectra and there was no evidence for specific interaction of protein HMG 1 with any of the subfractions. It therefore remains an open question as to whether protein HMG 1 and histone H1 are complexed together in chromatin.     

Larval adhesive organs and metamorphosis in ascidians

Summary The cup-shaped adhesive papillae of Distaplia occidentalis evert at the onset of metamorphosis and each transforms into a hyperboloidal configuration. The rate of transformation is a function of temperature. At 14° C complete eversion takes about 30 seconds. Myoepithelial cells that extend from the rim to the base on the cup contract. Simultaneously the central part of the papilla advances 60–70 m. During the last phases of eversion, collocytes (cells that secrete adhesives) on the inner wall of the cup and on the sides of the axial protrusion flow outward and form a collar-like structure.The myoepithelial cells contain arrays of thick and thin filaments. These become compacted during contraction. The surfaces of these cells become extensively folded as they shorten to about 1/3 of rest length. According to the proposed model the myoepithelial cells are the driving force in papillary eversion.Immediately after eversion is completed the papillae begin to retract. Eversion of the papillae is not inhibited by cytochalasin B, but the process of retraction is reversibly inhibited.Some histological characteristics of five types of everting papillae in four families of ascidians are compared.     

Structural changes occurring in interrenal tissue of the duck (Anas platyrhynchos) following adenohypophysectomy and treatment in vivo and in vitro with corticotropin

Adrenal glands from ACTH-treated intact ducks and chronically adenohypophysectomized ducks showed clear zonation into a subcapsular zone (SCZ) and an inner zone (IZ). Adenohypophysectomy caused ultrastructural changes in the IZ but not in the SCZ cells. These included increases in lipid droplets, changes in mitochondrial cristae from tubular to shelf-like, and changes in the shape of the nuclei from spherical to crenated. These changes were reversed by treatment with ACTH. Also, cells of the IZ, but not the SCZ, of adrenals from intact birds given ACTH showed more SER, more dense bodies, fewer lipid droplets and more prominent Golgi complexes. IZ cells incubated in buffer containing no ACTH developed mitochondria with shelf-like cristae and numerous opaque granules in the matrix. Exposure to buffer containing ACTH caused the mitochondrial cristae to become tubular and the matrix granules either decreased in number or disappeared. The granules could be extracted by incubating sections with chelating agents. The mitochondria in SCZ cells did not respond structurally to the presence of ACTH in the incubation medium but the matrix granules, like those in IZ cells, responded to the presence of chelating agents.