Sex behavior and hormone responses in ewes administered testosterone propionate

Two ewes were administered testosterone propionate and subsequent plasma testosterone concentrations determined and male sex behavior recorded. Initially ewes were administered 50 mg of testosterone propionate every other day for 20 days. Within 6 days following the first injection, concentrations of testosterone in plasma increased to 8.0 to 10.0 ng/ml. A 50 mg injection of testosterone propionate administered every 10 days thereafter maintained concentrations of plasma testosterone at 1.0 to 3.0 ng/ml. Sex behavior tests conducted with non-estrus and estrus ewes showed that both testosterone treated ewes developed male sex behavior similar to a ram. Ewes in estrus were mounted by testosterone treated ewes an average of 6.7 ± 1.2 times during a 10 minute test whereas none of the non-estrus ewes were ever mounted. Silastic implants containing testosterone propionate placed in the ewes 83 days following the first injection maintained concentrations of plasma testosterone at 6.0 to 8.0 ng/ml for a 20 day period. Therefore, administration of testosterone propionate to ewes effectively stimulates male sex behavior and would obviate the necessity for vasectomized rams for estrus detection.     

Pituitary desensitization and the regulation of pituitary gonadotropin-releasing hormone (GnRH) receptors following chronic administration of a superactive GnRH analog and testosterone

We recently demonstrated that chronic daily administration of a superactive GnRH analog to intact rats resulted in an initial stimulation of serum LH levels with a subsequent return of LH levels to baseline at a time when testosterone levels were marked decreased. These data demonstrated pituatary desensitization following chronic GnRH analog treatment. Administration of GnRH analog with a dose of testosterone which did not markedly lower serum LH levels when administered alone prevented the stimulation of LH secretion by analog. The present studies were undertaken to determine the effects of GnRH analog and testosterone administration on the regulation of pituitary GnRH receptors. Pituitary GnRH receptor binding was increased by analog treatment alone at 20 days and returned to control levels at 40 and 60 days of treatment in parallel to the observed changes in serum LH, demonstrating that one mechanism by which chronic GnRH analog treatment leads to pituitary desensitization is down-regulation of pituitary GnRH receptors. Testosterone administration alone decreased pituitary GnRH receptor binding. Combined GnRH analog and testosterone administration prevented the increase in pituitary GnRH receptors observed with analog administration alone. These studies demonstrate that changes in pituitary GnRH receptor binding correlate with changes in serum LH and that the stimulatory effects of analog administration on LH are sensitive to inhibition by small doses of testosterone.     

Physiological responses of two marine diatoms to pulsed additions of ammonium

The effects of pulsed ammonium additions on the ammonium-limited marine diatoms, Chaetoceros gracile Schutt and Skeletonema costatum (Grev.) Cleve were studied. Two culture systems were maintained for each species. One culture was grown in a chemostat which provided a homogeneous distribution of the limiting nutrient. In the other continuous culture, ammonium was added once daily giving rise to a patchy distribution. Ammonium patchiness increased both the V'_max (from 4.1 ± 0.3 to 11.8 ± 0.9 · h^?1) and the V_i (from 0.19 ± 0.02 to 0.36 ± 0 0.04 · h^?1) for ammonium in S. costatum and the V'_max (from 0.91 ± 0.07 to 2.9 ± 0 0.2 · h^?1) and the V_i (from 0.16 ± 0.01 to 0.31 ± 0.05 · h^?1) for ammonium in Chaetoceros gracile. The once-per-day addition of ammonium also induced a diel periodicity in photosynthetic rate and fluorescence although these cultures were growing under continuous light. The relative amplitude of the periodicity was greater for Skeletonema than for Chaetoceros. These observations are considered with regard to the hypothesis that limiting nutrient patchiness may alter the growth kinetics of marine phytoplankton.     

Growth hormone and prolactin secretion in Huntington's disease.

Growth hormone (GH) and prolactin (PRL) secretion was studied in twelve patients with Huntington's Disease, eight unaffected relatives, and twenty normal subjects in response to provocative and suppressive tests. Prolactin responses to TRH, chlorpromazine, L-DOPA, and apormorphine were similar in all groups with the exception of a slightly blunted PRL response to THR in the unaffected relatives. Although GH responses to L-DOPA were similar in all groups, patients with Hungtinton's Disease had nearly absent GH responses to apomorphine (mean peak GH = 1.4±0.4 (SE) ng/m1) compared to normal control subjects (mean peak GH = 28.9±8.6 ng/m1). These results, which are similar to some previously reported findings in drug-induced tardive dyskinesia, suggest an abnormality in dopamine-mediated GH secretion in Huntington's Disease.     

Reproductive hormone secretions and first service conception rate subsequent to ovulation control with Synchro-Mate B

Two groups of beef females receiving suboptimal energy diets were treated with Synchro-Mate B to control ovulation. The first group consisted of 30 suckled cows and 16 heifers. These females were bled 10 days and immediately prior to the implantation of norgestomet implants, at implant removal, 24, 27, 30, 33 and 36 hours and 9 and 16 days post-implant removal. The second group which consisted of 40 cows and 8 heifers was handled in the same manner except no blood samples were collected from 24 to 36 hours following implant removal. Calves were removed from all the cows for 48 hours, beginning at implant removal. All animals were artifically inseminated 48 hours following implant removal. Blood plasma was assayed for concentrations of progesterone and LH. The first service conception rate was 21% and 40% for groups 1 and 2. Several factors were identified that reduced the first service conception rate. In summary, Snychro-Mate B is an effective method to synchronize estrus in cattle. However, stress subsequent to implant removal should be avoided in order to obtain a higher first service conception rate.     

Guinea pig T-cell in vitro proliferative responses to tyrosine-azobenzenearsonate-pulsed and azobenzenearsonate-coupled macrophages have different specificities

The cell interactions involved in azobenzenearsonate-N-acetyl-tyrosine (ABA-tyr)-induced delayed hypersensitivity in the guinea pig were studied by in vitro blastogenesis. The ABA-sensitive lymphocyte was demonstrated to be a T lymphocyte and its presence in peritoneal exudate cells was shown to be much higher than spleen or lymph node populations. The secondary response of ABA-sensitized lymphocytes to ABA-tyr in culture is dependent on the presence of an accessory cell, with both splenic and peritoneal macrophages being equally effective. ABA coupled directly to macrophages as an immunogen induced strong responses to itself and not to ABA-tyr-pulsed macrophages or ABA-tyr in solution. The reverse was true in animals, immunized with ABA-tyr. ABA conjugated to thymocytes, L₂C leukemia cells, and guinea pig erythrocytes however, did not elicit significant responses. The results obtained in animals immunized with ABA- or ABA-tyr-modified cells was similar whether or not CFA was used. The difference in specificity shown between ABA-coupled and ABA-tyr-pulsed macrophages favors a single receptor hypothesis for T-cell recognition.     

Exposure to 4-aminopyridine prevents depressant effects of opiates on sensory-evoked dorsal-horn network responses in spinal cord cultures

The depressant effects of morphine (0.1-1 microM) on sensory-evoked dorsal-horn network responses in explants of mouse spinal cord with attached dorsal root ganglia (DRGs) were rapidly restored after addition of 4-aminopyridine (4-AP; 0.1 mM) and major components of these cord responses were stably maintained in the presence of the opiate. Moreover, prior exposure of cord-DRG explants to 0.1 mM 4-AP prevented the depressant effects of 0.1 microM morphine on DRG-evoked dorsal-horn responses, and the effects of 1-10 microM morphine were at least partly antagonized. Increased Ca++ levels (5 microM) attenuated the depression of dorsal horn responses by 1-10 micro M morphine and these effects of Ca++ were greatly enhanced in the presence of 4-AP--in some cultures, concentrations of morphine as high as 100 micro M were strongly antagonized during test periods up to 2 hours. Receptor assays showed that 0.1 mM 4-AP +/- 5 mM Ca++ had no effect on stereospecific opiate binding, indicating that the antagonist actions of these agents in our cultures do not occur at the level of the opiate receptor. The relevance of our in vitro studies of 4-AP antagonism of opiate-depressant effects on sensory-evoked dorsal-horn network responses for analyses of problems in opiate analgesia has been strengthened by a recent report demonstrating that 4-AP does, in fact, reverse morphine analgesia in rats, as determined by tail flick tests.     

Differentiation of the uterus in preparation for gestation: a model for the action of progesterone

During the preimplantation stages of pregnancy, rising titers of progesterone alter the metabolism of the uterine endometrium to permit implantation of the blastocyst. In this model of progestational differentiation, it is proposed that endometrial pyridine nucleotide metabolism is a key target of progestogen action. The hormone may modulate NAD metabolism to promote NADP synthesis while inhibiting NAD breakdown to ADP ribose and nicotinamide. The result of such an action would impair uterine DNA synthesis and cell division, but provide increased NADP for coenzyme-limited synthetic processes and cytodifferentiation. As a result, the endometrium differentiates and becomes sensitive to decidual-inducing stimuli (the blastocyst). The decidual stimulus reverses the process by rapidly inhibiting NADP production, and by dramatically increasing poly ADP ribosylation of nuclear protein, thus facilitating DNA synthesis and the wave of cell division associated with the initiation of decidualization. The background information and evidence in support of this model are presented.     

Purification of dog VIP from a single animal

VIP, a potent vasodilator peptide, is reported to be identical in pig, cow, human and rat but to differ in four amino acids in chicken. This report describes the purification of dog VIP from the small intestine of a single animal. The purification method is based on tissue extraction with a sequence of organic solvents. The extracted VIP is concentrated onto cation-exchange cellulose and brought to purity by three HPLC steps. A 30% final yield of pure VIP was obtained from the original extract. Dog VIP was found to have the following sequence: His-Ser-Asp-Ala-Val-Phe-Thr-Asp-Asn-Tyr-Thr-Arg-Leu-Arg-Lys-Gln-Met-Ala -Val-Lys-Lys-Tyr-Leu-Asn-Ser-Ile-Leu-Asn. Thus the amino acid sequence of dog VIP is identical with all the mammalian VIP's which have been reported. This suggests that a high degree of conservation throughout the molecule may be required for VIP bioactivity.     

The roles of the opioid peptides in controlling thyroid stimulating hormone release

We have studied the role of the opioid peptides in controlling TSH secretion. Morphine sulfate significantly decreased, while naloxone had no effect on, basal plasma TSH levels of female rats. In contrast, naloxone blocked the stress-induced fall in plasma TSH. Microinjection of β-endorphin into the third ventricle resulted in a fall in TSH while such injection of naloxone into the posterior hypothalamus increased TSH. Microinjection of β-endorphin directly into the pituitary caused a rise in plasma TSH. It is concluded that opioid peptides probably play no role in basal TSH secretion, but are involved in the stress-induced fall in TSH. Furthermore, it appears that opioid peptides have a site of action in the hypothalamus to decrease TSH and a direct pituitary action to increase TSH.     

Effect of leucogenenol,a thymothyroid hormone,on the enzymes and several other constituents of the serum

It has been found that rats treated with the thymothyroid hormone, leucogenenol, have significantly elevated levels of alkaline phosphatase and lactate dehydrogenase in their serum 8 hrs. later. These levels remain elevated for 24 hrs. and 12 hrs., respectively. The concentrations of creatine phosphokinase and aspartate aminotransferase as well as total protein, albumin, calcium, inorganic phosphorus, glucose, urea nitrogen, uric acid and creatinine in the serum are not affected by treatment with leucogenenol. These results are in agreement with the previously reported findings that treatment with leucogenenol increases the rate of development of blood cells of the bone marrow, increases the rate of recovery from immunosuppression and enhances the immune response.     

Dopa decarboxylase activity in Aedes aegypti: A preadult profile and its subsequent correlation with ovarian development

Dopa decarboxylase activity was monitored throughout the entire life cycle of Aedes aegypti. Peaks of activity were detected at each larval molt, at the larval-pupal ecdysis, and at eclosion. The dopa decarboxylase activity in adults was high right after eclosion, but it then dropped rapidly and after 5 days very little activity was detectable. This activity, however, was persistent and remained essentially constant, albeit low, for up to 15 days of adult life. Throughout this part of the study no sex differences in enzymatic activity were observed.A dramatic increase in the level of dopa decarboxylase was noted after adult females were allowed to blood feed. Since a blood meal is necessary in order to initiate ovarian development in this species and since the rate of increase of enzymatic activity paralleled oocyte maturation a causal relationship was indicated. Specifically, we suggest that the dopa decarboxylase is incorporated into the eggs to be used later for subsequent sclerotization.Injection of the molting hormone β-ecdysone into non-blood fed females resulted in a marked stimulation of dopa decarboxylase activity. No such stimulation was observed in saline-injected adult females. The adult female enzymatic activity profile obtained with time after hormone injection was qualitatively the same as that seen after a blood meal. The possibility that ecdysone or an ecdysonelike hormone is necessary for normal ovarian development in Aedes aegypti is discussed.     

Dissociation between increased growth hormone and prolactin secretion during the morning hours of early pregnancy in the rat

We investigated whether serum growth hormone (GH) concentration changes in association with the rise in serum prolactin (PRL) concentration known to occur during the early morning hours in the pregnant rat. Animals were kept in a room with the lights on from 0500 to 1900 hours (hr) daily and decapitated for the collection of trunk blood at 2200 or 2400 hr on Day 6 of pregnancy or at 0200, 0400, 0800 or 1000 hr on Day 6 of pregnancy. Serum GH concentration rose more than 4-fold from low levels at 2200 and 2400 hr to higher levels at 0400 and 0800 hr and then declined by 1000 hr. Serum prolactin (PRL) concentration followed a similar pattern except that it returned to low levels earlier, by 0800 hr. Serum luteinizing hormone, follicle-stimulating hormone and thyroid-stimulating hormone concentrations showed no significant changes. Serum GH levels at 0800 hr in pregnant rats were higher than those observed in cyclic rats (13 time periods sampled). The results demonstrate that serum GH concentration is elevated during a circumscribed period in the 6- to 7-day pregnant rat. The time of onset of the rise is similar to that for serum PRL but the elevation in GH levels persists longer than that for PRL.     

Bacteria-derived human growth hormone lacks lipolytic activity in rat adipose tissue

Bacteria-derived human growth hormone (hGH) shows little $\text{in}\text{vitro}$ lipolytic activity in adipose tissue from fed rats. In adipose tissue from fasted rats no lipolytic activity is observed. However, bacteria-derived hGH increased serum free fatty acids after intraperitoneal administration to hypophysectomized rats to the same extent as purified pituitary hGH. The dose response of the bacteria-derived hGH tested for $\text{in}\text{vitro}$ insulin-like activity was very similar to the pituitary extracted material. Thus bacteria-derived hGH behaves in a manner indistinguishable from highly purified preparations of pituitary hGH.     

Thyrotropin-releasing hormone and a homologous peptide in the reproductive system of the female rat and pig

TRH and a TRH homologous peptide have been shown to occur throughout the female rat and pig reproductive systems by TRH radioimmunoassay, SP-Sephadex C-25 cation exchange chromatography, and parallel line analysis of the assays. The total amount of TRH and TRH homologous peptide immunoreactivity was highest in the oviducts followed by the ovary and then uterus. The concentration of TRH immunoreactivity in all reproductive organs of the rat fell gradually from one month of age. TRH and the TRH homologous peptide were not parallel on serial dilution and measurement in the same TRH radioimmunoassay. The rapid degradation of TRH by pig follicular fluid may explain the higher measured concentration of TRH homologous peptide compared to TRH not only in pig follicular fluid but also in the pig ovary as a whole.     

Association of leucogenenol,a thymothyroid hormone,with carrier proteins in the thymus

Leucogenenol a heterocyclic enolic thymothyroid hormone (MW 383) whose concentration in the serum regulates the rate at which already committed cells of the bone marrow develop into functional cells, was found to be associated in the thymus with a carrier protein. The carrier protein for leucogenenol is not precipitated by heating to 80° but following this treatment leucogenenol is precipitated in association with proteins precipitated by acetone and then by saturated ammonium sulfate. On chromatography on Sephacryl G-200 it was found that leucogenenol was associated with proteins of MW approximately 38,000. Leucogenenol is not eluted from the chromatographic column if it is not associated with its carrier proteins. It is suggested that other hormones such as those associated with the reproductive cycle or compounds that result from tissue damage induce the liberation of leucogenenol from its carrier protein in the thymus to the circulation where it is associated as previously described, with a protein of approximately MW 300,000.     

Species-specific dissociation into single cells of live sea urchin embryos by fab against membrane components of Paracentrotus lividus and Arbacia lixula

The species and stage specificities of membrane components active in promoting reaggregation of cells dissociated from embryos of the two Mediterranean sea urchin species Paracentrotus lividus and Arbacia lixula have been examined. Membrane proteins extracted with butanol either from purified membranes or from dissociated cells without significant reduction of viability promoted reaggregation of both the homologous and heterologous species. Extracts from plutei and blastulae were equally effective in promoting reaggregation of blastula cells. By contrast, Fab's prepared from IgG raised against these extracts or purified membranes are strictly species specific because they prevent reaggregation of cells and actively dissociate live embryos of only the homologous species. No corresponding stage specificity of the Fab was observed: Fab against extracts from blastula embryos also caused dissociation of plutei. Antigenic analysis of the extracts by the Ouchterlony test revealed the presence of components specific for each species as well as others common to both.     

Multiple bands produced by interaction of a single macromolecule with carrier ampholytes during isoelectric focusing

Equilibrium isoelectric focusing patterns have been computed for reversible, carrier ampholyte-induced macromolecular isomerization reactions. The calculations predict that an amphoteric macromolecule, interacting with n species of ampholyte located at different positions along the isoelectric focusing column, can give a pattern showing n + 1 well-resolved peaks under appropriate conditions.     

Age-related alterations in plasma membrane glycoprotein content and scheduled or unscheduled DNA synthesis.

WI-38 cells of various ages and SV40-transformed WI-38 cells were examined for differences in plasma membrane composition of glycoproteins and DNA synthesis. Sialic acid per milligram of protein content of the membranes of WI-38 cells decreased with passage of time in culture. Other glycoprotein fractions and alkaline phosphatase activity disappeared in the WI-38 cells with passage of time in culture (Phase III). Studies of DNA repair correlated with changes observed in the plasma membrane glycoprotein content of WI-38 cells over a passage of time in culture were also reported. Both the extent and rate of ultraviolet-induced unscheduled DNA synthesis remained relatively constant during the passage of the WI-38 cells until late phase III. At that time the extent of unscheduled DNA synthesis was measurably reduced. The number of cells in a population of phase III cells able to perform semiconservative DNA synthesis diminished with age in culture but not to an extent capable of explaining the observed changes seen in membrane composition of semiconservative DNA synthesis during passage of the cells in culture. Cells with an extended lifespan SV40-transformed WI-38 (VA 13.2 RA) cells, did not vary in membrane composition, semiconservative DNA synthesis, or unscheduled DNA synthesis over 200 serial subpassages of the cells in culture.