A single protein catalyzes both N-deacetylation and N-sulfation during the biosynthesis of heparan sulfate

Heparan sulfate is a highly sulfated carbohydrate polymer that binds to and modulates the activities of numerous proteins. The formation of these protein-binding domains in heparan sulfate is dependent on a series of biosynthetic reactions that modify the polysaccharide backbone; the initiating and rate-limiting steps of this process are the N-deacetylation and N-sulfation of N-acetylglucosamine residues in the polymer. We now report that in the rat liver, biosynthesis of heparan sulfate utilizes a single protein that possesses both N-deacetylase and N-sulfotransferase activities. This was accomplished by demonstrating that both activities resided in a purified soluble fusion protein containing the Golgi-lumenal portion of the enzyme. We propose that this protein be renamed the rat liver Golgi heparan sulfate N-deacetylase/N-sulfotransferase.     

Evidence for a role for the cyclic adenosine 3',5'-monophosphate/protein kinase-A pathway in regulation of the gonadotropin subunit messenger ribonucleic acids

cAMP regulation of gonadotropin secretion and subunit mRNA levels was studied in pituitary cells perifused with pulses of GnRH. Pituitary cells from 7-week-old male rats castrated at 5 weeks of age were stimulated hourly for 9-24 h with 1-min pulses of GnRH, the adenylate cyclase activator forskolin, the cell-permeable cAMP analog 8-bromo-cAMP (8Br-cAMP), or control medium. Cells were also treated with the nonsteroidal antiinflammatory drug flufenamic acid, which reduces pituitary cAMP levels. During perifusion, the effluent was collected in 10-min fractions for FSH and LH assay. At the completion of perifusion, total RNA was extracted, and gonadotropin subunit mRNA levels were quantitated by Northern analysis. Continuous administration of flufenamic acid gradually reduced the amplitude of GnRH-stimulated FSH and LH pulses to nadir values of 40 +/- 4.7% and 62 +/- 12% of the control value, respectively. Flufenamic acid decreased (P < 0.05) FSH beta and alpha-subunit mRNA levels and blocked the effect of GnRH to lengthen LH beta mRNA. Pulses of forskolin or 8Br-cAMP released LH and FSH, and continuous forskolin or 8Br-cAMP potentiated the gonadotropin stimulatory effect of GnRH. Forskolin or 8Br-cAMP increased (P < 0.05) FSH beta mRNA and alpha-subunit mRNA levels when administered in pulses, but not when administered continuously, and lengthened LH beta mRNA. The Nal-Glu GnRH antagonist blocked the effects of GnRH pulses, but not the effects of 8Br-cAMP or forskolin. In conclusion, lowering intracellular cAMP levels with flufenamic acid attenuated GnRH-stimulated gonadotropin secretion, decreased alpha-subunit and FSH beta mRNA levels, and blocked the effect of GnRH to lengthen LH beta mRNA, whereas 8Br-cAMP or forskolin produced the opposite effect. These data extend previous results which suggested that cAMP modulates gonadotropin secretion and indicate that the cAMP/A-kinase pathway regulates each of the gonadotropin subunit mRNAs.     

Chloramphenicol antibody causing interference in antibody detection and identification tests

Investigation of the serum of three patients with positive antibody detection tests demonstrated the cause in each to be an antibody against chloramphenicol, a bacteriostatic agent used in commercial red blood cell reagents. Washing of these red cells prior to use prevented agglutination. All three examples of anti-chloramphenicol antibody were IgM and were in low titer when tested at room temperature and 37 C in saline. Two of the antibodies bound complement. The possibility of an antibody to an ingredient of the commercial preservative solution should be considered if problems are encountered in tests with unwashed commercial red blood cell reagents.     

Hemodynamic effects of thiothixene and chlorpromazine in schizophrenic patients at rest and during exercise

The hemodynamic effects and plasma levels of noradrenaline were studied in schizophrenic patients at rest and during exercise after long-term treatment with chlorpromazine (150-600 mg daily) and thiothixene (60-80 mg daily). The results are compared with those from previous studies in untreated patients and patients receiving very large doses of chlorpromazine. The effects of thiothixene on the different hemodynamic variables were very moderate, and the observed differences between this group and the control group may be due to the different patient materials. In the two groups of patients receiving chlorpromazine, the heart rate at rest and durng exercise tended to be higher than in the control group. There was also a tendency towards a lower stroke volume after this drug and thiothixene during exercise. The noradrenaline levels in plasma were highest after the high dose of chlorpromazine both at rest and during exercise, while they were lower after the moderate chlorpromazine dose. After thiothixene, the values were between those of the group on the low chlorpromazine dose and those of the control group.     

Messenger RNA synthesis, transport, and storage in silkmoth ovarian follicles

The effect on liver microsomal enzyme activity of three steroid contraceptive drug (SCD) combinations was compared in rats, mice and guinea-pigs. Lynestrenol plus mestranol, norethisterone plus mestranol and norethynodrel plus mestranol were given orally for 4 consecutive days (acute treatment) or 30 days (chronic treatment) at various doses eliciting an experimentally controlled antifertility activity which varied in its extent. In rats and mice all the combined treatments (with the exception of norethynodrel plus mestranol in mice) were active as inducers of liver microsomal enzymes. This induction seems to be mediated mainly by the progestogenic compounds. Oestrogens showed a very poor effect bordering on significance only in a few cases. No effect on liver microsomal protein or cytochrome P 450 concentration was obtained after treatment with doses capable of increasing the microsomal enzyme activity. The activity of the liver microsomal enzymes did not appear to be reduced immediately (2 h) after the last administration of the SCD given during 4 or 30 days. Contraceptive treatments at doses capable of eliciting complete antifertility activity were inactive on liver microsomal enzyme activity in guinea-pigs.     

Preintervention effects of mediation versus arbitration.

Examined the effects of the anticipation of various forms of 3rd-party intervention in a collective bargaining simulation. 50 male graduate students served as ss. Results indicate negotiators faced with a binding decision from a 3rd party behaved in a more conciliatory fashion and were more likely to reach agreement than those faced with a nonbinding decision. This effect was more significant for union negotiators than management negotiators. (PsycINFO Database Record (c) 2010 APA, all rights reserved)