Modulation of lipolysis by endotoxin and indomethacin in fat cells of germfree and conventional dogs.

Germfree fat cells released significantly less FFA and glycerol under basal conditions (i.e. in the absence of hormonal stimulation) than conventional cells. The lipolytic response to norepinephrine stimulation (0.2 μg/ml) was not different in the two cell populations.$\text{E. coli}$ endotoxin increased basal lipolysis and norepinephrine stimulated (0.2 μg/ml) FFA release in adipocytes from conventional dogs, while having no consistent influence on lipolysis of adipocytes from germfree dogs. The endotoxin effect was not dose dependent (0.2–2.0 μg/0.5 ml cell suspension).Indomethacin (5.0 μg/ml) significantly increased basal FFA and glycerol release from cells of germfree origin, and FFA efflux from cells of conventional dogs. Endotoxin obviated the influence of indomethacin on basal lipolysis of germfree cells.Endotoxin by itself did not alter cAMP concentrations in adipocytes from germfree dogs. The combination of indomethacin and endotoxin, however, significantly increased intracellular cyclic nucleotide concentrations.Compared to conventional fat cells, germfree fat cells are characterized by significantly reduced basal lipolysis, lack of a consistent lipolytic response to endotoxin stimulation and dissociation of the lipolytic response and cAMP levels by the combined influence of endotoxin and indomethacin.     

Pregnancy rate in peripuberal beef heifers following treatment with syncro-mate-B and gonadotropin releasing-hormone

A study was conducted to evaluate the effectiveness of treatment with Syncro-Mate-B (SMB) and gonadotropin releasing-hormone (GnRH) on inducing a fertile estrus and/or ovulation in peripuberal beef heifers. Two hundred and twenty-three, 13 to 15 month peripuberal beef heifers, primarily of Angus or Simmental breeding, were alternately allotted by weight and age within breed to a non-treated control group (C) and three groups which received the SMB treatment regime. Treated groups differed in that one group was inseminated approximately 12 hr after detected in estrus (T) while two other groups were inseminated at a fixed time after implant removal (T-G-I42, T-I48). Heifers in group T-G-I42 received 125 mug GnRH at 30 hr, and were inseminated 42 hr after implant removal. The remaining group of SMB-treated heifers received no additional hormone therapy but were inseminated 48 hr after implant removal (T-I48). Age had no significant effect on estrous response or pregnancy rate nor were there significant age by weight interactions on these parameters. Weight was a significant source of variation on interval from start of breeding to pregnancy (P<.01) and the effect on interval to estrus approached statistical significance (P<.0.9). In control heifers, increased weight had a positive effect on estrous response during the first 21 days of breeding (P<.05). Conversely, weight had no effect on estrous response during the 5-day synchronized period in heifers treated with SMB and observed for estrus (group T). Cumulative estrous response in control heifers was 10, 47, 48 and 55% in 5, 21, 27 and 45 days of breeding, respectively. Cumulative estrous response in SMB-treated heifers obsrved for estrus (group T) was 88, 91, 91 and 91% in 5, 21, 27 and 45 days of breeding, respectively. Pregnancy rate after 5 days of breeding was 9, 27, 18 and 30% for heifers in groups C, T, T-G-I42 and T-I48, respectively. Pregnancy rate after 21 days of breeding for heirfers in groups C, T, T-G-I42 and T-I48, respectively, was 33, 38, 21 and 41%; after 27 days of breeding was 36, 43, 40 and 48%; after 45 days of breeding was 48, 61, 51 and 69% and after the total 88-day breeding season was 59, 68, 71 and 72%. While a large percentage of peripuberal beef heifers with adequate age but insufficient weight were induced into estrus with the SMB treatment, pregnancy rates were low. Timed insemination programs with or without GnRH do not appear to be effective in improving pregnancy response in non-cyclic beef heifers.     

d-Ribulose 1,5-bisphosphate carboxylase and polyhedral inclusion bodies in Nitrosomonas spec

Polydedral inclusion bodies were isolated from exponentially grown cells of Nitrosomonas spec. The bodies contained d-ribulose, 1,5-bisphosphate carboxylase. The specific activity of the enzyme was 0.0122 mol CO₂ fixed per min per mg of protein.     

Cloning and characterization of the CYS3 (CYI1) gene of Saccharomyces cerevisiae.

A DNA fragment containing the Saccharomyces cerevisiae CYS3 (CYI1) gene was cloned. The clone had a single open reading frame of 1,182 bp (394 amino acid residues). By comparison of the deduced amino acid sequence with the N-terminal amino acid sequence of cystathionine gamma-lyase, CYS3 (CYI1) was concluded to be the structural gene for this enzyme. In addition, the deduced sequence showed homology with the following enzymes: rat cystathionine gamma-lyase (41%), Escherichia coli cystathionine gamma-synthase (36%), and cystathionine beta-lyase (25%). The N-terminal half of it was homologous (39%) with the N-terminal half of S. cerevisiae O-acetylserine and O-acetylhomoserine sulfhydrylase. The cloned CYS3 (CYI1) gene marginally complemented the E. coli metB mutation (cystathionine gamma-synthase deficiency) and conferred cystathionine gamma-synthase activity as well as cystathionine gamma-lyase activity to E. coli; cystathionine gamma-synthase activity was detected when O-succinylhomoserine but not O-acetylhomoserine was used as substrate. We therefore conclude that S. cerevisiae cystathionine gamma-lyase and E. coli cystathionine gamma-synthase are homologous in both structure and in vitro function and propose that their different in vivo functions are due to the unavailability of O-succinylhomoserine in S. cerevisiae and the scarceness of cystathionine in E. coli.     

Gelation of xanthan with trivalent metal ions

In-situ gelation of semidilute xanthan solutions with trivalent chromium, aluminum or iron ions was studied by rheology and UV-spectroscopy. Measurements of the elastic modulus of xanthan gel cylinders prepared by dialysis against the complexing ion at pH values from 2 to 6 indicate that monomeric species of the ion are ineffective, whereas dimeric or higher oligomeric species are effective in crosslinking the polysaccharide. When chromium was used as the crosslinking species, the dependence of the gelation rate on the ionic concentration followed a power law with a coefficient of 1·7. The gelation time and the gelation rate were found to extrapolate to zero at 1 m Cr for 2·5 mg/ml xanthan. The limiting concentration of xanthan needed for gelation with 5 m Cr(III) at 20°C was estimated as 0·35 mg/ml. This critical xanthan concentration is close to the overlap concentration c^* estimated from the experimentally determined intrinsic viscosity [η] using c^* = 1·4/[η]. An apparent activation energy for crosslinking of xanthan was calculated as E_a = 42 kJ/mol and E_a = 108 kJ/mol for Cr and Al ions, respectively. The fractal dimensionality of xanthan-Cr at the sol-gel transition was estimated as 1·3 applying the Chambon-Winter criterion for gelation, thus indicating that this gelation criterion is applicable also to stiff-chain polysaccharides such as xanthan.     

Role of calcium and protein kinase C in development of the delayed rectifier potassium current in Xenopus spinal neurons.

The delayed rectifier current of embryonic Xenopus spinal neurons plays the central role in developmental conversion of calcium-dependent action potentials to sodium-dependent spikes. During its maturation, this potassium current undergoes a pronounced increase in rate of activation. The mechanism underlying the change in kinetics was analyzed with whole-cell voltage clamp of neurons cultured under various conditions. Calcium is necessary at an early stage of development, to permit influx that triggers subsequent release of calcium from intracellular stores. Its action is prevented by depletion of protein kinase C and mimicked by stimulation of the kinase. Calcium influx through voltage-dependent channels at early stages of development regulates the differentiation of potassium current kinetics and modulation of the ionic dependence of action potentials.     

Down-regulation of prostaglandin F2 alpha receptors in rat liver during chronic endotoxemia.

Prostaglandin (PG) F2 alpha binding parameters were measured in purified plasma membrane preparations isolated from livers of chronically endotoxin-(ET) treated rats and corresponding controls. Two classes of binding sites were detected in both groups: high affinity, low capacity, with a KD of 44.4 +/- 8.8 nM for saline- and 28.6 +/- 11.3 nM for ET-treated rats (n = 5 for both, p greater than 0.05) and low affinity, high capacity with a KD of 1.12 +/- 0.49 microM for saline- and 1.24 +/- 0.43 microM for ET-treated rats (p greater than 0.05). Bmax values for high affinity sites were 1.01 +/- 0.18 fmol.mg-1 protein for saline- and 1.02 +/- 0.54 (same units) for ET-treated rats (p greater than 0.05). There was a significant difference (p less than 0.01) between the Bmax values for low affinity sites in saline- (675 +/- 332 fmol.mg-1 protein) and ET-treated rats (12 +/- 1, same units). This decrease in the amount of PGF2 alpha low affinity high capacity binding sites may underlie the depression of the PGF2 alpha stimulatory effect on hepatic gluconeogenesis induced by non-lethal, chronic ET treatment of rats, recently described by us (9).     

Response of anestrous ewes to norgestomet and PMSG

A 10-day treatment regime with a subcutaneous ear implant containing 3 mg of norgestomet, accompanied by an intramuscular injection of 1.5 mg norgestomet and 0.5 mg estradiol valerate (EV) on day 1 and 750 I. U. pregnant mares serum gonadotropin (PMSG) given intravenously on day 10, proved effective in eliciting estrus in 72% of 110 anestrous ewes within 5 days of treatment. Ewes which were treated in months closer in proximity to the normal breeding scason responded with significatly increased induction of estrus, with 71, 37, 59, 74, and 97% in estrus for ewes which were treated in February through June, respectively. Comparable estrous response in nontreated, control ewes was 0, 13, 0, 10, and 24% during February through June, respectively. (Treated vs controls, P<.01). Pregnancy rate to first service of ewes in estruc was 51% in treated and 30% in control ewes (P>.10). Overall pregnancy rate for all ewes in both groups was 36% in treated and 3% in control ewes during 5 or 16 days of breeding, respectively (P<.01).