"Pertussis toxin induces tachycardia and impairs the increase in blood pressure produced by alpha 2-adrenergic agonists"

Administration of purified pertussis toxin to rats induced persistent tachycardia, (observed in conscious rats but not after pithing); as little as 0.05 microgram/100 g produced a significant effect. Pertussis toxin-treatment did not affected the pressor response produced in the pithed rats by the alpha 2-adrenergic agonist methoxamine but markedly diminished the pressor effect of the alpha 2-adrenergic agonists clonidine and azepexole. A role of adenylate cyclase inhibition in the action of postsynaptic vascular alpha 2-adrenergic receptors is suggested.     

Protein-bound sulfhydryl groups and thiolesters in mitochondria and submitochondrial particles and their relationships to oxidative phosphorylation

Procedures are described for measurement of total protein-bound —SH groups and apparent thiolester linkages in mitochondrial preparations. The procedure for total —SH is based on reaction with radioactive p-mercuribenzoate in 6 m guanidine hydrochloride at pH 9 containing 1 m ammonia buffer. The procedure for apparent thiolesters is based on alkylation of available — SH groups in 6 m guanidine hydrochloride, protein precipitation with perchloric acid, and ammonolysis at pH 9 to liberate — SH from any thiolester present.No variations in total —SH groups measured in Submitochondrial particles or in mitochondria were noted with presence or absence of ATP, ADP, substrates, or uncouplers of oxidative phosphorylation. Previous reports of changes in measured — SH groups thus appear to reflect solely changes in the reactivity of — SH groups and not in the total — SH groups present.Submitochondrial particles contained less than 0.5 nmole of apparent thiolester per mg of protein; mitochondria contained 1 to 2 nmoles. No variation of apparent thiolester content was noted with change in conditions affecting oxidative phosphorylation. The data do not, of course, eliminate possible changes in thiolester content below the sensitivity of the detection procedures.     

The excretion pattern of biliverdin and bilirubin in bile of the small skate (Raja erinacea)

Summary Bile pigment composition (biliverdin, bilirubin and their conjugates) was analyzed in stored gallbladder bile and newly synthesized hepatic bile from the small skate (Raja erinacea). During a five day period of captivity, gallbladder volume remained relatively constant while bilirubin and biliverdin content increased two to three fold. Biliverdin which accounted for 50% of the pigments did not increase as a percentage of tetrapyrroles during this period. The relative proportion of bilirubin and its conjugates also remained constant, averaging 65% for bilirubin monoglucuronide, 30% for bilirubin diglucuronide and 5% for unconjugated bilirubin as measured by HPLC methods. Intravenous administration of biliverdin resulted in significant increases in the biliary excretion of both biliverdin and all bilirubin tetrapyrroles. Insignificant quantities of³H-biliverdin were detected in hepatic bile following the intravenous administration of³H-bilirubin. These studies indicate that the small skate excreted both biliverdin and bilirubin conjugates in bile and that the biliverdin was not produced by in vitro oxidation of bilirubin or its metabolites.     

Specific Inhibition of Lignification in Bryonia dioica: Effects on Thigmomorphogenesis

Rubbing-induced lignification in Bryonia dioica internodes is significantly impaired by N(o-hydroxyphenyl) and N(o-aminophenyl) sulfinamoyl tertiobutyl acetate, specific inhibitors of cinnamyl alcohol dehydrogenase, an enzyme which is strictly associated with lignin monomer synthesis. Along with the reduction of lignification, these inhibitors counteract the inhibition of elongation due to rubbing. These results indicate that lignification participates in the thigmomorphogenetic growth response of Bryonia dioica internodes. In a general way, the data point to the causal role of lignification in the limitation of plant growth.     

Clustering of null mutations in the EcoRI endonuclease

EcoRI endonuclease mutants were isolated in a methylase-deficient background following in vitro hydroxylamine mutagenesis of plasmid pKG2 (Kuhn et al.: Gene 44:253-263, 1986). Mutants which survived high-level endonuclease expression (IPTG induction) were termed null mutants. Sixty-two of 121 null mutants tested by Western blot contained normal levels of endonuclease cross-reacting protein. The complete endonuclease gene was sequenced for 27 null mutants. This group was found to consist of 20 single base-change missense mutations, 6 double mutations, and 1 triple mutation. Ten of the 20 single mutations were clustered between residues 139 and 144. When examined with respect to the structure of the EcoRI-DNA complex (McClarin et al.: Science 234:1526-1541, 1986), these alterations were found to fall predominantly into two classes: substitutions at the protein-DNA interface or substitutions at the protein-protein (dimer) interface. Protein from several of the mutants was purified and sized by using HPLC. Wild-type EcoRI endonuclease and protein from three of the DNA interface mutations (Ala139----Thr, Gly140----Ser, Arg203----Gln) appeared to be dimeric, while protein from subunit interface mutations (Glu144----Lys, Glu152----Lys, Gly210----Arg) migrated as monomers.     

Failure of familiar males to prevent alien male-induced implantation block (the Bruce effect) in laboratory mice

Implantation failure in newly inseminated females induced by exposure to alien males (the Bruce effect) was significantly reduced when the females were housed with the stud male. By contrast, newly inseminated females housed with a familiar male during exposure to alien males exhibited a high rate of implantation failure. The results suggest that the protective effect of the stud male on implantation is not because of the familiarity of the female with his odour cues. The results are consistent with the view that the newly inseminated female mouse identifies her coital partner as an individual because she becomes 'imprinted' with his odour during the pericopulatory period.     

Purification and characterization of the restriction endonuclease RsrI, an isoschizomer of EcoRI

Rhodobacter sphaeroides strain 630 produces restriction enzyme RsrI which is an isoschizomer of EcoRI. We have purified this enzyme and initiated a comparison with the EcoRI endonuclease. The properties of RsrI are consistent with a reaction mechanism similar to that of EcoRI: the position of cleavage within the -GAATTC-site is identical, the MgCl2 optimum for the cleavage is identical, and the pH profile is similar. Methylation of the substrate sequence by the EcoRI methylase protects the site from cleavage by the RsrI endonuclease. RsrI cross-reacts strongly with anti-EcoRI serum indicating three-dimensional structural similarities. We have determined the sequence of 34 N terminal amino acids for RsrI and this sequence possesses significant similarity to the EcoRI N terminus.