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When the amount of activation of ribulose 1,5-bisphosphate carboxylase has been measured, two forms of the enzyme, not one, are actually determined experimentally. Only the enzyme-activator CO(2)-Mg(2+) form can bind ribulose bisphosphate for reaction with substrate CO(2) or O(2). A method is presented which measures only this catalytically active form by stabilizing it with ribulose bisphosphate just before dilution and assay in Mg(2+)-free reaction medium.

Basal adenylate cyclase values for corpora lutea (CL) removed from cyclic gilts on Days 3, 8, 13 and 18 were 178 +/- 61, 450 +/- 46, 220 +/- 25 and 208 +/- 18 pmol cAMP formed/min/mg protein, respectively. Basal activity was significantly elevated on Day 8 (P less than 0.001). LH-stimulatable adenylate cyclase values for CL from Days 3, 8, 13 and 18 were 242 +/- 83, 598 +/- 84, 261 +/- 27 and 205 +/- 17 pmol cAMP formed/min/mg protein respectively. Serum progesterone concentrations of 12 gilts bled every 2 days through one complete oestrous cycle ranged from 1.1 to 26.9 ng/ml with highest values between Days 8 and 12. The decline in serum progesterone concentrations was coincident with the decrease in basal adenylate cyclase activity. There was no LH-stimulatable adenylate cyclase activity present in the CL at the specific times of the oestrous cycle examined. We conclude that progesterone secretion by the pig CL is apparently dependent on basal activity of adenylate cyclase.

Male bluegill (Lepomis macrochirus) display a complex reproductive behavior involving two alternative life history pathways: delay of sexual maturation to become "parentals" or precocious maturation as "cuckolders." The purpose of our study was to investigate the association of two androgens, 11-ketotestosterone (11KT) and testosterone (T), with reproduction in these two types of males. Radioimmunoassay techniques were used to measure daily levels of the two androgens in the blood serum of parental male bluegill captured during the prespawning, spawning, and nesting periods throughout the reproductive season. Dramatic changes in the levels of 11KT and T were observed among parental males during these periods. Peaks occurred at the onset of spawning activity during each breeding bout. Compared to spawning parental males, spawning cuckolder males had significantly lower serum levels of 11KT. In contrast, the serum levels of T among parental and cuckolder males were not significantly different. These findings suggest that the elevated levels of 11KT are associated with the behaviors displayed by spawning parental males. The levels of T, however, seem to be associated with the occurrence of a phenomenon common to both parental and cuckolder males, such as development of gonads and/or spermiation.

Holstein cows just past peak lactation were used in a 3 x 3 Latin square design to determine the effects of arginine infusion on concentrations of somatotropin and insulin in plasma, milk production, and milk composition. Treatments were: 1) control; 2) arginine injection into jugular vein, and 3) arginine infusion into abomasum. Concentrations of arginine and ornithine in plasma were increased by injection of arginine into the jugular vein compared with the control. The concentration of ornithine in plasma was increased shortly after injection of arginine into the jugular vein, and both arginine and ornithine concentrations in plasma decreased rapidly. Abomasal infusion of arginine significantly increased concentrations of arginine, ornithine, and urea in plasma compared with concentrations in the control treatment. Injection of arginine into the jugular vein increased concentrations of somatotropin and insulin in plasma, but the increase did not persist for more than 30 min. A secondary peak in plasma somatotropin concentration occurred approximately 1 h after the initial peak. Arginine infusion into the abomasum did not alter plasma concentrations of either somatotropin or insulin. Dry matter intake, milk production, and milk composition were not affected by treatments. Lack of changes in milk production and milk composition suggest that acute increases in somatotropin with concomitant increases in insulin are not sufficient to stimulate synthesis of milk and milk components by cows during established lactation.

"An attempt is made in the light of an analysis of population changes occurring in several sub-districts of Greater Santiago in the period between the last population censuses of 1970 and 1982, based on small-area census units (distritos and communas) to draw attention to the effects on the spatial growth of the city resulting from changed conditions in the political and juridical sphere since the change of power in 1973." The effect of changes in government policy concerning the liberalization of the property market and the illegal occupation of land are noted. (summary in ENG), (excerpt)

In the golden hamster drugs which inhibit monoaminergic function (including p-chlorophenylalanine (PCPA), methysergide and a-methyl-p-tyrosine (a-MPT) facilitated lordosis in ovariectomized female hamsters as a function of the duration of estradiol benzoate (EB) priming. a-MPT (200 mg/kg), methysergide (6 mg/animal) or PCPA (150 or 360 mg/kg) potentiated lordosis if 6 days of EB priming preceded drug treatment. However, if female hamsters were primed with EB for only 2 days, a-MPT and methysergide were ineffective. PCPA (360 mg/kg) was less effective after 2 days of EB than after 6 days of EB priming. alpha-MPT produced a three-fold elevation in progesterone levels in ovariectomized females but methysergide and PCPA did not influence serum progesterone. PCPA (360 mg/kg) facilitated lordosis in adrenalectomized, ovariectomized females, eliminating the possibility that adrenal progesterone is essential for the behavioral effects of the drug. Luteinizing hormone-releasing hormone levels in the preoptic/anterior hypothalamic area and the medial basal hypothalamus were also not significantly altered at 1 h after PCPA injection. Pimozide (1.5 mg/kg) and pimozide (1.5 mg/kg) and amphetamine (2.5 mg/kg) did not potentiate lordosis in ovariectomized hamsters after either 2 or 6 days of EB priming. Pargyline, a monoamine oxidase inhibitor, inhibited female sexual behavior in females in estrogen alone-induced estrus. Lordosis in the female rat is more readily elicited both by drugs and estrogen. It is proposed with regard to female sexual behavior that species differences in estrogen sensitivity may underlie apparent differences in drug sensitivity.

Previous studies have shown that the rabbit fetal placenta contains both biological and immunological gonadotrophin-releasing hormone- (GnRH-) like activity. The purposes of this study were to determine whether (1) the fetal placenta secretes a GnRH-like factor, and (2) this GnRH-like factor is secreted into the uterine vein. In experiment 1, fetal placentae were collected from day-18 pregnant rabbits. The placentae were divided in half, and one-half was used for measurement of GnRH; the other half was cut into small pieces and placed into a perifusion system. Following a 2-h stabilization period, tissue was challenged with two 60 mM K+ injections spaced 90 min apart. Six replicates were run with either liver (n = 4) or spleen (n = 2) as control tissues. Basal levels of GnRH secreted (nmol/g protein/30 min) were 1.2 +/- 0.3 for placental tissue, 0.3 +/- 0.1 for liver tissue and non-detectable for spleen tissue. Following the first K+ injection there was no response from liver or spleen tissue, while placental tissue showed a 78 per cent increase after 90 min. There was no response to the second injection. The concentration of GnRH in the placenta was 6.8 +/- 0.5 nmol/g protein. In experiment 2, plasma samples were obtained from the marginal ear vein of five oestrous does and five day-7 pseudopregnant does, and from the marginal ear vein and uterine vein of six day-18 pregnant does.(ABSTRACT TRUNCATED AT 250 WORDS)

The purpose of the present investigation was to measure the concentrations of progesterone (P4), testosterone (T) and estradiol-17 beta (E2) in isolated theca and granulosa layers of the five preovulatory follicles of the domestic hen. The largest follicle (F1), the second largest (F2), third largest (F3), fourth largest (F4) and fifth largest (F5) follicles were removed at 24, 18, 12, 6 and 2 h before the expected ovulation. Theca and granulosa layers were isolated and P4, T, E2 and protein concentrations determined. Protein concentrations of the granulosa and theca layers increased 5- and 15-fold, respectively, during the five ovulatory cycles prior to ovulation. As the follicle approached ovulation, there was a linear decrease in E2 concentration of the theca layer with the most significant decrease (P less than 0.001) occurring between 24 and 18 h of the ovulatory cycle. In the F3 and F4 theca layers, there was a significant increase (P less than 0.005) in E2 at 6 h of the ovulatory cycle. Fluctuations in T concentrations in theca and granulosa layers were similar. There was a significant increase (P less than 0.05) in T in both layers of the F2, F3 and F4 follicles at 6 h followed by a decrease (P less than 0.005) in the theca layers at 2 h of the ovulatory cycle. The P4 concentration of the granulosa layer increased gradually during follicular maturation, with the greatest increase occurring in the F2 and F1 granulosa layers between 18 and 12 h of the ovulatory cycle.(ABSTRACT TRUNCATED AT 250 WORDS)

The ovulatory cycle of the domestic hen is approximately 26 h in length. The hen ovulates an egg each day at a progressively later time until she finally skips a day, resets her "clock" and a new sequence is started. The ovarian component of this unique timing mechanism is the focus of this report. In Experiment 1, we asked whether there was a difference in luteinizing hormone (LH)-stimulated progesterone (P4) secretion by the granulosa layer removed from the largest follicle (F1) that had been the F1 follicle for 8, 12, or 32 h. In Experiment 2, our objective was to determine whether the theca layer of an F1 follicle influenced P4 secretion by the granulosa layer of that follicle and whether such an interaction depended on the maturity of the F1 follicle (had been a F1 follicle for 8 h or 32 h). Results from Experiment 1 revealed that there was no significant difference in LH-stimulated P4 secretion by the granulosa layer in a perifusion system regardless of the length of time the follicle had been the largest follicle. In contrast, in Experiment 2, when granulosa and theca layers from the same follicle were co-incubated in a perifusion system, P4 secretion from the more mature F1 follicle (32 h) increased in response to LH, whereas P4 secretion from the less mature F1 follicle (8 h) was not elevated by LH.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of initial oxygen concentration on the rate and extent of oxidation of the respiratory chain carriers of anaerobic mitochondria from mung bean (Phaseolus aureus) seedlings was examined. The substrate was succinate, with malonate added to give malonate to succinate ratios of 6 to 12, thereby minimizing the flow of reducing equivalents from substrate and insuring maximal extent of oxidation of the carriers. The ratio of oxidizing equivalents available from oxygen to reducing equivalents available from reduced ubiquinone, designated the equivalents ratio, varied from 30 to 1. Cytochromes aa(3) and c(547) have unaltered oxidation half-times, designated t((1/2) on), as the equivalents ratio is reduced from 30 to 3, and the extent of oxidation is decreased by about 25%. The time of the oxidation-reduction cycle induced by the oxygen pulse, calculated from the point of half oxidation to that of half reduction and designated t((1/2) off), decreases 200 fold with this reduction in equivalents ratio. The oxidation half-time, t((1/2) on), for ubiquinone is unaltered by decreasing the equivalents ratio from 6 to 1; the value of t((1/2) off) decreases only 30% while the extent of oxidation decreases 50%. The values of t((1/2) on) and t((1/2) off) and the extent of oxidation of cytochrome b(553) and flavoprotein Fp(ha) were all much reduced at low equivalents ratios. The results, plus results from previous studies, indicate that there is the following linear sequence of components in the plant respiratory chain:Dehydrogenase.UQ.Fp(ha).b(553). c(549).a.a(3)Cytochrome b(557) and the low potential flavoproteins do not fit into this linear sequence.

Hydroxamic acids, R-CONHOH, are inhibitors specific to the respiratory pathway through the alternate, cyanide-insensitive terminal oxidase of plant mitochondria. The nature of the R group in these compounds affects the concentration at which the hydroxamic acids are effective, but it appears that all hydroxamic acids inhibit if high enough concentrations are used. The benzhydroxamic acids are effective at relatively low concentrations; of these, the most effective are m-chlorobenzhydroxamic acid and m-iodobenzhydroxamic acid. The concentrations required for half-maximal inhibition of the alternate oxidase pathway in mung bean (Phaseolus aureus) mitochondria are 0.03 mm for m-chlorobenzhydroxamic acid and 0.02 mm for m-iodobenzhydroxamic acid. With skunk cabbage (Symplocarpus foetidus) mitochondria, the required concentrations are 0.16 for m-chlorobenzhydroxamic acid and 0.05 for m-iodobenzhydroxamic acid. At concentrations which inhibit completely the alternate oxidase pathway, these two compounds have no discernible effect on either the respiratory pathway through cytochrome oxidase, or on the energy coupling reactions of these mitochondria. These inhibitors make it possible to isolate the two respiratory pathways and study their mode of action separately. These inhibitors also enhance an electron paramagnetic resonance signal near g = 2 in anaerobic, submitochondrial particles from skunk cabbage, which appears to be specific to the alternate oxidase and thus provides a means for its assay.

The kinetics of oxidation of ubiquinone, flavoprotein, cytochrome c, and the cytochrome b complex in skunk cabbage (Symplocarpus foetidus) mitochondria made anaerobic with succinate have been measured spectrophotometrically and fluorimetrically in the absence of respiratory inhibitor and in the presence of cyanide or antimycin A. No component identifiable by these means was oxidized rapidly enough in the presence of one or the other inhibitor to qualify for the role of alternate oxidase. Cycles of oxidation and rereduction of flavoprotein and ubiquinone obtained by injecting 12 mum oxygen into the anaerobic mitochondrial suspension were kinetically indistinguishable in the presence of cyanide or antimycin A, implying that these 2 components are part of a respiratory pathway between succinate and oxygen which does not involve the cytochromes and does involve a cyanide-insensitive alternate oxidase. The cytochrome b complex shows biphasic oxidation kinetics with half times of 0.018 sec and 0.4 sec in the absence of inhibitor, which increase to 0.2 sec and 1 sec in the presence of cyanide. In the presence of antimycin A, the oxidation of the cytochrome b complex shows an induction period of 1 sec and a half-time of 3.5 sec. A split respiratory chain with 2 terminal oxidases and a branch point between the cytochromes and flavoprotein and ubiquinone is proposed for these mitochondria.

Mitochondria were prepared from the spadices of skunk cabbage (Symplocarpus foetidus) whose respiratory rate with succinate and malate showed 15% to 30% sensitivity to cyanide inhibition, and which showed respiratory control by added ADP. The observed respiratory control ratios ranged from 1.1 to 1.4. The change in pH of the mitochondrial suspension was recorded simultaneously with oxygen uptake: alkalinization of the medium, expected for phosphorylation of ADP, coincided with the period of acceleration in oxygen uptake caused by addition of an ADP aliquot. The ADP/O ratios obtained were 1.3 for succinate and 1.9 for malate. In the presence of 0.3 mm cyanide, the ADP/O ratio for succinate was zero, while that for malate was 0.7. These results are consistent with the existence of an alternate oxidase which interacts with the flavoprotein and pyridine nucleotide components of the respiratory chain and which, in the presence of cyanide, allows the first phosphorylation site to function with an efficiency of about 70%. In the absence of respiratory inhibitors, the efficiency of each phosphorylation site is also about 70%. This result implies that diversion of reducing equivalents through the alternate oxidase, thereby bypassing the 2 phosphorylation sites associated with the cytochrome components of these mitochondria, occurs to a negligible extent during the oxidative phosphorylation of ADP or State 3. Addition of ADP or uncoupler to skunk cabbage mitochondria respiring in the controlled state or State 4, results in reduction of cytochrome c and the oxidation of the cytochromes b, ubiquinone and pyridine nucleotide. A site of interaction of ADP with the respiratory chain between cytochromes b and cytochrome c is thereby identified by means of the crossover theorem. Flavoprotein measured by fluorescence is also oxidized upon addition of ADP or uncoupler, but flavoprotein measured by optical absorbance changes becomes more reduced under these conditions. Depletion of the mitochondria by pretreatment with ADP and uncoupler prevents reduction of most of the fluorescent flavoprotein by succinate. These results indicate that skunk cabbage mitochondria contain both high and low potential flavo-proteins characterized by different fluorescence/absorbance ratios similar to those demonstrated to be part of the respiratory chain in mitochondria from animal tissues.