Modulation of xanthine dehydrogenase and oxidase activities during the hormonal induction of vaginal epithelial differentiation in ovariectomized mice

In situ hybridization and immunocytochemical techniques have been used to examine the distribution of vitamin-D-induced calbindin mRNA and calbindin protein in enterocytes lining the crypts and villi of chicken small intestine. Basal villus enterocytes contained approximately twice as much calbindin but over three times as much calbindin mRNA compared to values found in basal crypt and upper villus enterocytes, all values being measured 2 days after vitamin D injection into D-deficient chickens. Virtually no calbindin mRNA was detected in tissues taken from control D-deficient birds. Direct proportionality found between calbindin mRNA and calbindin content in enterocytes of basal crypt, mid and upper villus suggests pre-translational control over calbindin synthesis. The implications of possible inefficient translation of calbindin mRNA in basal villus enterocytes are discussed. Present methods of analysis provide a novel way to study mechanisms controlling gene expression throughout the whole process of enterocyte differentiation.     

Multispecific DNA methyltransferases from Bacillus subtilis phages. Properties of wild-type and various mutant enzymes with altered DNA affinity

Temperate Bacillus subtilis phages SPR, phi 3T, rho 11 and SP beta code for DNA methyltransferases, each having multiple sequence specificities. The SPR wild-type and various mutant methyltransferases were overproduced 1000-fold in Escherichia coli and were purified by three consecutive chromatographic steps. The stable form of these multispecific enzymes in solution are monomers with a relative molecular mass (Mr) of about 50,000. The methyl-transfer kinetics of the SPR wild-type and mutant enzymes were determined with DNA substrates carrying either none or one of the three recognition sequences (GGCC, CCGG, CCATGG). Evaluation of the catalytic properties for DNA and S-adenosylmethionine binding suggested that the NH2-terminal part of the protein is important for both non-sequence-specific DNA binding and S-adenosylmethionine binding as well as transfer of methyl groups. On the other hand, mutations in the COOH-terminal part lead to weaker site-specific interactions of the enzyme. Antibodies raised against the purified SPR enzyme specifically immunoprecipitated the phi 3T, rho 11 and SP beta methyltransferases, bu failed to precipitate the chromosomally coded enzymes from B. subtilis (BsuRI) and B. sphaericus (BspRI). Immunoaffinity chromatography is an efficient purification step for the related phage methyltransferases.     

Cytosine-specific DNA modification interferes with plasmid establishment in Escherichia coli K12: involvement of rglB

Summary Several chimeric pBR322/328 derivatives containing genes for cytosine-specific DNA methyltransferases (Mtases) can be transformed into the Escherichia coli K12/E. coli B hybrid strains HB101 and RR1 but not into other commonly used E. coli K12 strains. In vitro methylation of cytosine residues in pBR328 and other unrelated plasmids also reduces their potential to transform such methylation sensitive strains, albeit to a lesser degree than observed with plasmids containing Mtase genes. The extent of reduced transformability depends on the target specificity of the enzyme used for in vitro modification. The role of a host function in the discrimination against methylated plasmids was verified by the isolation of K12 mutants which tolerate cytosine methylated DNA. The mutations map in the vicinity of the serB locus. This and other data indicate that the host rglB function is involved in the discrimination against modified DNA.     

Topographic control of vegetation in a mountain big sagebrush steppe

Mountain big sagebrush steppes in Wyoming have strong spatial patterning associated with topography. We describe the spatial variability of vegetation in a sagebrush steppe, and test the relationship between topography and vegetation using canonical correlation. Results of the analysis suggest that the main control over vegetation distribution in this system is wind exposure. Exposed sites are characterized by cushion plant communities and Artemisia nova, and less exposed sites by the taller sagebrush species Artemisia tridentata ssp. vaseyana. Topographic depressions and leeward slopes are characterized by aspen stands and nivation hollows. Measurements of soil microclimate suggest that a major influence of topographic position on vegetation is snow redistribution and its effect on soil moisture and temperature.Abbreviations ARNO Artemisia nova - ARTRW Artemisia tridentata ssp. wyomingensis - ARTRV Artemisia tridentata ssp. vaseyana - PUTR Purshia tridentata - RIP riparian community - POTR Populus tremuloides - NIV nivation hollow community     

Planar asymmetric lipid bilayers of glycosphingolipid or lipopolysaccharide on one side and phospholipids on the other: membrane potential, porin function, and complement activation.

We have determined some physicochemical properties of the monosaccharide-type fraction (GSL-1) of glycosphingolipids, the major glycolipid components of the outer leaflet of the Gram-negative species Sphingomonas paucimobilis. These properties included the state of order of the hydrocarbon moiety, the effective molecular area, surface charge density, and intrinsic transmembrane potential profile of reconstituted planar asymmetric GSL-1/phospholipid bilayer membranes. We have, furthermore, investigated the insertion into and the function of porin channels in the reconstituted bilayers and the complement-activating capability of GSL-1 surfaces. All results were compared with respective data for deep rough mutant lipopolysaccharide of Salmonella minnesota R595. We found a remarkable agreement in most functional properties of the two glycolipids.     

Nitrate limitation of N2O production and denitrification from tropical pasture and rain forest soils

Nitrous oxide production was measured in intact cores taken from active pasture and old-growth forest Inceptisols in the Atlantic Lowlands of Costa Rica. Following additions of aqueous KNO₃ or glucose, or the two combined amendments, the cores were incubated in the laboratory to determine if N₂O production rates were either N-limited or C-limited in the two land use types. Differences in rates of denitrification (N2₂O + N₂ production) among amended forest and pasture soils were determined by addition of 10% C₂H₂.The forest soils were relatively insensitive to all amendment additions, including the acetylene block. Forest N₂O production rates among the treatments did not differ from the controls, and were consistently lower than those of the pasture soils. With the addition of glucose plus nitrate to the forest soils, production of N₂O was three times greater than the controls, although this increase was not statistically significant. On the other hand, the pasture soils were definitely nitrogen-limited since N₂O production rates were increased substantially beyond controls by all the amendments which contained nitrate, despite the very low N level (5 mg N kg^–1 soil) relative to typical fertilizer applications. With respect to the nitrate plus glucose plus acetylene treatment, denitrification was high in the pasture soils; N₂O production in the presence of C₂H₂ was 150% of the rate of N₂O production measured in the absence of the acetylene block. The results are discussed in relation to the effects of agricultural land use practices and subsequent impacts of disturbance on N₂O release.