Auxin-induced H+-pump stimulation does not depend on the presence of epidermal cells in corn coleoptiles

Cell-wall acidification and electrical reactions (depolarization and hyperpolarization) are typical auxin responses in maize (Zea mays L.) coleoptiles. In an attempt to test the role of the outer epidermis in these responses, they have been measured and compared in intact and peeled coleoptile fragments. To exclude interactions between parenchymal and epidermal cells, the coleoptile pieces were completely stripped of their outer epidermis. This preparation was monitored by means of a scanning electron microscope. When externally applied indole-3-acetic acid was tested, we found that neither cell-wall acidification nor the electrical membrane responses depended on the presence of intact epidermal cells.Abbreviations IAA Indole-3-acetic acid - MES 2-[N morpholino-ethane-sulfonic acid - TRIS 2-Amino-2-hydroxymethyl-1,3-propanediol We thank Kuki Kaethner for her excellent technical assistance. This work was supported by the Hessische Graduiertenförderung and the Deutsche Forschungsgemeinschaft.     

A high-affinity bradykinin receptor in membranes from rat myometrium is coupled to pertussis toxin-sensitive G-proteins of the Gi family

In rat myometrial membranes, two 3H-Bradykinin binding sites with KD values of 16 pM and 1.0 nM were identified. Employed at pM concentrations, bradykinin stimulated high affinity GTPases. This effect was abolished by the bradykinin antagonist, [D-Arg(Hyp3-Thi5,8, D-Phe7)]bradykinin (10 microM), and by treatment of membranes with pertussis toxin. Myometrial membranes contained two pertussis toxin substrates of 40 and 41 kDa, which corresponded immunologically to alpha-subunits of Gi-type G-proteins. The faster migrating substrate was tentatively identified as Gi2 alpha-subunit. The electrophoretic mobility of the slower migrating Gi alpha-subunit was very similar to that of the Gi3 alpha-subunit. Go alpha-subunits were not detected. Thus, in uterine smooth muscle, G-proteins of the Gi-family (Gi2, Gi3) couple high-affinity bradykinin receptors to their effector enzymes.     

Mapping of mglB, the structural gene of the galactose-binding protein of Escherichia coli

Summary The tetracycline resistance transposon Tn10 was inserted into the E. coli chromosome near mglB550, a structural gene for the galactose-binding protein. P1 transductions established the position of these Tn10 insertions (zee-700, 701, 702::Tn10) close to the genes ptsF, fpk, cdd, mglB550, his, and gatA with 85%–95%, 85%, 36%, 20%–40%, 12%–15%, and 0.5% contransduction frequency. Three factor crosses revealed the relative sequence of the genes as: mglB550, zee-700::Tn10, ptsF, fpk, cdd, his. gatA was found to be 1.3% cotransducible with mglB550. Two Tn10 insertions near gatA were isolated and characterized. One, zef-704::Tn10, was 3% cotransducible with fpk, 8% with mglB550, and 42% with gatA. The other, zef-703::Tn10, was 98% cotransducible with gatA but not with mglB550 or fpk. Neither of these two Tn10 insertions was cotransducible with cdd. Four factor crosses revealed the sequence gatA, zef-704::Tn10, mglB550, fpk.Neither zee-700::Tn10 nor zef-703::Tn10 showed any (0/300) contransduction with either glpT or gyrA. The clockwise order of genes is then: his, cdd, fpk, ptsF, zee-700::Tn10, mglB550, zef-704::Tn10, gatA. With a fix-point for his at 44 min, fpk would be placed at 45 min and mglB550 at 45.5 min. During the course of this work we noticed that the cotransduction frequency between Tn10 insertions and nearby markers tended to increase when new P1 lysates were prepared from freshly reisolated strains. This may indicate loss of nonessential genes adjacent to Tn10 insertions. Using insertion zee-702::Tn10, we isolated deletions extending into an mgl gene other than mglB. Crosses between such a deletion mutant and an mglB550 mutant were done. The analysis of the periplasmic proteins of these as well as other transductants or recombinants involving the mglB550 or the mglB551 gene revealed the existence of strains synthesizing both the wild-type as well as the corresponding mutant protein. Strains containing both proteins exhibit either wild-type or mutant phenotype. These strains appeared unstable. Upon reisolation from purified stock cultures kept in glycerol at-20°C, colonies could be isolated that carried only mutant or wild-type protein.     

Binding of non-substrate nucleotides to a restriction endonuclease: a model for the interaction of bam HI with its recognition sequence

The kinetic constants of the site-specific endonuclease Bam HI for various substrates were determined and binding of non-substrate nucleotides to the enzyme was studied. Agarose gel assays in combination with an integrated Michaelis-Menten equation were used for the evaluation of data. The turnover number was 2.2 min-1 at 37 degrees C with pJC80 DNA as the substrate. It depends on the conformation and base composition of the substrate. Michaelis constants also depend on substrate conformation. Non-substrate polynucleotides were found to inhibit Bam competitively with KI ranging from 10(-6) to > 10(-3) M depending on base composition, base pairing, and helix conformation. Dinucleotides showed sequence-specific, competitive inhibition with KIs ranging from 10(-5) to > 10(-3) M. Mononucleotides and -nucleosides acted noncompetitively. Binding was influenced by the extent of phosphorylation, but not by the nature of the base. KIs varied between 10(-3) and 10(-2) M. The results are discussed with respect to the recognition requirements of Bam HI.     

Peroxidase cytochemistry and ultrastructure of resident macrophages in fetal rat liver: A developmental study

The peroxidase cytochemistry and the ultrastructural characteristics of resident macrophages in fetal rat liver have been investigated. Livers of 10-, 11-, 14-, 17-, and 20-day-old fetuses were fixed by immersion or perfusion, incubated for peroxidase, and processed for transmission electron microscopy. Some 17- and 20-day-old fetuses were injected prior to sacrifice with carbon or 0.8-μm latex particles through the umbilical vein. Some livers were additionally processed for scanning electron microscopy (SEM). The endogenous peroxidase was present in the nuclear envelope (NE) and endoplasmic reticulum (ER) of fetal macrophages with a negative reaction in the Golgi apparatus, a distribution pattern identical to that in Kupffer cells of adult rat liver. Such peroxidase-positive cells avidly took up the injected latex and carbon particles and were the only cell type in fetal liver involved in erythrophagocytosis. Furthermore, they were associated with erythropoietic elements, forming close contacts with such cells, especially normoblasts. The peroxidase pattern in leukopoietic cells differed at all stages of maturation from that in macrophages. By SEM the macrophages exhibited ruffles and lamellopodia on their surfaces and protruded often into the lumen of fetal sinusoids. Macrophages in fetal liver underwent mitotic divisions. The macrophages were first seen on gestation day 11, whereas the first mature monocytes were found on gestation day 17. These observations suggest that resident macrophages in fetal rat liver form a self-replicating cell line independent of the monocytopoietic series, although they may both arise from a common precursor cell.     

Efficacy of different types of aerobic exercise in fibromyalgia syndrome: a systematic review and meta-analysis of randomised controlled trials

Introduction  

The efficacy and the optimal type and volume of aerobic exercise (AE) in fibromyalgia syndrome (FMS) are not established. We therefore assessed the efficacy of different types and volumes of AE in FMS.     

Proteolytic enzymes in Chlamydomonas I. A survey on the aminopeptidase pattern in asynchronous vegetative cells of Chlamydomonas reinhardii

The supernatant of a crude extract from vegetative cells ofChlamydomonas reinhardii contains three different types of aminopeptidases.They are similar in their substrate specificities to the relativealanine specific aminopeptidases, the relative leucine specificaminopeptidases and the specific proline iminopeptidases describedin many other systems. Relative alanine specific aminopeptidasewhich also cleaves N-terminal Lys and Leu residues has a molecularweight of 92,000 daltons and is inhibited by zinc and manganeseions.Relative leucine specific aminopeptidase shows high activitywith N-terminal Phe besides Leu, and is capable of cleavingTyr, Pro, and to a minor degree Ala. It has a molecular weightof 76,000 daltons. No effects on its activity were detectedin the presence of divalent cations or chelating agents. Theiminopeptidase specifically splits N-terminal Pro and has amolecular weight of about 255,000 daltons. All the enzymes showoptimal activity at pH 8.0–8.5. The two aminopeptidases can be separated from the iminopeptidaseby ammonium sulfate solubilization and from each other by subsequentfractionation on DEAE-cellulose. Relative leucine specific activityappeared as a single enzyme in all the fractionation techniquesused, but it gave two distinct bands when crude extracts wererun on native polyacrylamide gels. Therefore, this enzyme mayexist in multiple molecularforms. (Received October 17, 1978; )