Anterior Pituitary Extract Research Articles

Biosynthesis in vitro of immunoreactive 31,000-dalton corticotropin/β-endorphin-like material by bovine hypothalamus

Enzymatically dispersed bovine hypothalamic or cortical tissue was maintained in culture in the presence of (3)H-labeled amino acids. After such incubation, extracts of cells and of media contained (3)H-labeled products that were specifically bound by immobilized affinity-purified antisera to corticotropin (ACTH) and beta-endorphin. The majority of these products eluted in the void volume (V(0)) upon Sephadex G-50 gel filtration; minor (3)H-labeled products eluted in the regions of the ovine beta-lipotropin marker and in fractions having apparent molecular weights of approximately 12,000 and 3800. Sequential use of these immobilized antisera revealed that most of the V(0) material contained both ACTH and beta-endorphin antigenic determinants within the same molecule(s), whereas retarded material contained only one of the determinants. When this V(0) material was rerun on a Sephadex G-75 column, it coeluted with the 31-kilodalton precursor of both ACTH and beta-endorphin obtained from a bovine anterior pituitary extract. Thus, the high molecular weight immunoreactive ACTH/beta-endorphin-like (3)H-labeled product(s) derived from the hypothalamic culture is similar to the pituitary-derived precursor in containing the dual antigenic determinants and in its gel filtration characteristics. In contrast, the cortex-derived cell preparation was devoid of (3)H-labeled products specifically reactive with the antisera employed.

RELATIVE ACTIVITY, PLASMA ELIMINATION AND TISSUE DEGRADATION OF SYNTHETIC LUTEINIZING HORMONE RELEASING HORMONE AND CERTAIN OF ITS ANALOGUES

The abilities of three nonapeptide analogues of synthetic luteinizing hormone releasing hormone (LH-RH) to release luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in anoestrous and cyclic ewes were examined, as were their elimination from the plasma in vivo and degradation by extracts of the hypothalamus, anterior pituitary gland, lung, kidney, liver and plasma in vitro. In all cases, comparisons were made with synthetic LH-RH. When injected i.v. into mature ewes as a single dose, the potencies of the analogues were graded and Des Gly-NH2(10) LH-RH ethylamide was found to be the least potent. It was not possible to demonstrate any significant increase in the potency of this analogue over LH-RH, although a trend was apparent with each parameter examined. [D-Ser(But)6] Des Gly-NH2(10) LH-RH ethylamide had the greatest potency. There were no differences between the responses of anoestrous ewes and those of ewes treated on day 10 of the oestrous cycle. None of the analogues had a rate of elimination from the plasma different from that of LH-RH during either the first or the second components of the biphasic disappearance curve. The incubation of LH-RH with tissue extracts showed that extracts of the hypothalamus and anterior pituitary gland degraded LH-RH to a similar extent. Both the hypothalamic and anterior pituitary gland extracts degraded more LH-RH than did lung extract, which in turn destroyed more LH-RH than did extracts of kidney or liver tissue. The degradative abilities of kidney and liver extracts did not differ from each other. Plasma failed to degrade LH-RH or the analogues. Although LH-RH was rapidly destroyed by hypothalamic extract in vitro, of the analogues, only Des Gly-NH2(10) LH-RH ethylamide was degraded. The anterior pituitary gland and kidney extracts failed to degrade [D-Ser6] Des Gly-NH2(10) LH-RH ethylamide and [D-Ser(But)6] Des Gly-NH2(10) LH-RH ethylamide as rapidly as LH-RH. Extracts of liver and lung were incapable of catabolizing any of the analogues. There was an inverse correlation between the LH- and FSH-releasing potency of an analogue and its rate of degradation by anterior pituitary gland extract. The slower rates of catabolism of certain analogues of LH-RH by the anterior pituitary gland may explain their increased LH- and FSH-releasing potency.

Embryo transfer in the Angora goat

A total of 711 embryos was collected from 121 Angora does mated to Angora bucks. They were transferred to 667 feral recipient does, of which 361 kidded, producing 378 kids. Donors were treated with 36, 40 or 45 mg of an equine anterior pituitary extract (HAP) or 1500 i.u. pregnant mare serum gonadotrophin (PMSG) given at the end of a period of progesterone treatment (12 mg/day by intramuscular injection), while the recipients were either untreated, or their time of oestrus was controlled by progestagen-impregnated intravaginal pessaries or by the prostaglandin analogue 'Estrumate' (I.C.I.). All 121 donors exhibited oestrus after treatment, and 104 (86%) were in oestrus 48–60 h after the final injection of progesterone. Donors were naturally mated by allowing them free access to bucks, hand-mated at 12-h intervals, surgically inseminated into the uterus, or inseminated into the cervix; and in does treated with HAP the proportion of recovered eggs fertilized were respectively 88, 79, 65 and 32%. Seven does received PMSG. They were all hand-mated, and 46% of eggs recovered were fertilized. The mean numbers of corpora lutea in does treated with HAP and PMSG were 10.4 ± 0.9 and 13.7 ± 2.2, and it is suggested that the low rate of fertilization following PMSG was due to an excessive ovulatory response. Treatment with pessaries and Estrumate effectively controlled the time of oestrus in recipients and, overall, there was little difference between the proportions of treated and untreated recipients which kidded (58 v. 50% recipients kidded). Embryos were recovered from donors 3½–5½ days after they were first observed in oestrus, and they were transferred to recipients first observed in oestrus 48 h before to 48 h after their respective donors. The kidding rate of does which received day 3½ embryos was less than that of those which received older embryos (28 v. 55%), but the degree of asynchrony between respective donors and recipients had no effect upon survival.

Content of β-LPH and its fragments (including endorphins) in anterior and intermediate lobes of the bovine pituitary gland

Radioimmunoassays (RIAs) specific for β-LPH 1–47, β-endorphin, α-MSH and β-MSH have been used to identify immunoreactive components in acid extracts from anterior and intermediate lobes of bovine pituitary gland after separation by chromatography on Sephadex G-50. When components in extracts of both lobes, eluting at the same position, were measured with the β-endorphin and β-LPH 1–47 RIA systems, marked quantitative differences were seen. The main components reacting with the β-LPH 1–47 system in anterior pituitary extract co-migrated with β-LPH and γ-LPH while in the intermediate lobe, the main immunoreactive component eluted at a position slightly later than β-endorphin. When the β-endorphin RIA system was used, relatively low amounts of immunoreactive material co-migrating with β-endorphin were seen in the anterior lobe extract while a highly predominant peak eluting at a position slightly later than β-endorphin was observed in intermediate lobe extract. Some β-MSH was seen in the intermediate lobe. These date indicate that the processing of β-LPH is markedly different in the anterior and intermediate bovine pituitary lobes: β-endorphin immunoreactive material predominates in the intermediate lobe whereas β-LPH and γ-LPH predominate in the anterior lobe.

Existence of a common precursor to ACTH and endorphin in the anterior and intermediate lobes of the rat pituitary.

Extracts of rat anterior and intermediate-posterior pituitary were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and assayed for immunoactive ACTH and endorphin. In both lobes the major forms of immunoactive ACTH have apparent molecular weights of 31,000 (31K), 20--21K, 14K, and 4.5K, and the major forms of immunoactive endorphin have apparent molecular weights of 31K (coincident with the peak of immunoactive ACTH), 13K (a betaLPH-like peptide), and 3.5K (a beta-endorphin-like peptide). However, the quantitative distribution of immunoactivity among the various forms differs greatly between the lobes. Assays using an extreme COOH-terminal ACTH antiserum indicate that the 31K ACTH/endorphin molecule in rat anterior and intermediate pituitary is similar to the pro-ACTH/endorphin molecule from mouse pituitary tumor cells. A radioimmunoassay that is specific for the NH2-terminal non-ACTH, nonendorphin segment (referred to as 16K fragment) of the mouse pro-ACTH/endorphin molecule was used to assay extracts of rat pituitary. In addition to detecting material at 31K and 20--21K, the 16K fragment radioimmunoassay detects significant amounts of cross-reactive material with an apparent molecular weight of 16K in extracts of both lobes. This result also suggests that the structure and processing of the rat 31K ACTH/endorphin molecule is similar to that of mouse tumor cell pro-ACTH/endorphin. Cell suspensions were prepared from the anterior and intermediate lobes of the rat pituitary and maintained in culture for a 24-h period. The isolated cells from both lobes incorporate [3H] phenylalanine into immunoprecipitable ACTH- and endorphin-containing molecules. By sequential immunoprecipitation with ACTH and endorphin antisera, it is possible to demonstrate directly that a single molecule (31K ACTH/endorphin) has antigenic determinants for both ACTH and endorphin. Significant amounts of 31K ACTH/endorphin are released into the culture medium by isolated anterior lobe and intermediate lobe cells. The isolated intermediate lobe cells synthesize and secrete relatively large amounts of a beta-endorphin-like molecule; the isolated anterior lobe cells secrete significant amounts of both a betaLPH-like molecule and a beta-endorphin-like molecule. These same quantitative differences between anterior and intermediate lobe tissue were observed in immunoassays of extracts of the separated lobes and probably reflect differences in the processing of the common precursor. The isolated anterior lobe cells can be stimulated to release increased amounts of immunoprecipitable ACTH and endorphin by incubation with a cyclic AMP analog and a phosphodiesterase inhibitor.

Massive liver enlargement accompanied by decreased drug metabolism. Effect of anterior pituitary extract on hepatic ultrastructure, zoxazolamine paralysis, and metabolism in the rat

The relationship between liver enlargement and drug metabolism was investigated in female rats. Hepatomegaly (e.g., 31% increase in liver weight in a 17-day experiment) was induced by injection of lyophylized anterior pituitary (LAP) extract. The liver enlargement seemed to be due to an increase in the number and the size (enhanced water content and PAS-positive material) of hepatocytes. Electron microscopic examination of the liver revealed slight proliferation of the smooth endoplasmic reticulum and pronounced fragmentation and dilation of the rough endoplasmic reticulum. Zoxazolamine paralysis time was significantly prolonged (+55% and +102%) after 4 and 17 days, respectively, of treatment with LAP. Metabolism of zoxazolamine by the 9000 g supernatant fraction of the liver of rats given LAP for 17 days was reduced by 73%. Thus, the marked hepatomegaly induced by LAP was associated with a prolonged action of the drug which may result from a decrease in hepatic drug metabolism.

The Effects of Whole- and Partial-Body Irradiation on Circulating Anterior Pituitary Hormones and Testosterone and the Relationship of These Hormones to Drug-Metabolizing Enzymes in the Liver

McTAGGART, J., AND WILLS, E. D., The Effects of Whole- and Partial-Body Irradiation on Circulating Anterior Pituitary Hormones and Testosterone and the Relationship of These Hormones to Drug-Metabolizing Enzymes in the Liver. Radiat. Res. 72, 122-133 (1977). Doses within the range of 850 to 1500 rad given to the whole body, head, or lowertrunk region of male rats cause a marked depression in the rate of oxidative demethylation of drugs in the liver endoplasmic reticulum, 3 to 4 days after the irradiation. The Vmax of the enzyme system is depressed and the Km increased. Irradiation to the whole body, head, or lower trunk also causes a fall in the circulating levels of testosterone and luteinizing hormone (LH) and the concentrations of these hormones are markedly reduced 3 to 4 days after the irradiation. Injections of testosterone or anterior pituitary extract effectively restore the activity of the liver enzyme system after irradiation of the head or lower trunk. It is concluded that whole-body irradiation causes inhibition of drug-metabolizing enzymes in the liver by a complex series of interrelated effects on the

Studies on the Biological Properties and Gel Filtration Behavior of Pituitary and Serum Follicle Stimulating Hormone of Infantile Female Rats1

Molecular distribution profiles for FSH in anterior pituitary extracts or serum of infantile female rats (10–12 days of age), as determined by exclusion chromatography, were compared to those of prepubertal (27 days) or adult (70 days) rats. The elution position of pituitary FSH at the different ages studied was essentially the same, indicating the absence of major differences in average apparent molecular size. Characterization of serum FSH revealed a preponderance of FSH components of larger molecular size. However, the elution position of serum FSH was similar at all ages studied. Small quantitative differences were found between pituitary FSH potencies assessed by RIA or by bioassay. However, at the two ages studied (12 and 27 days) the immunoassay/bioassay ratios were similar. Pituitary FSH of both infantile and older female rats showed a dose-response curve in the ovarian hCGaugmentation assay parallel to that of NIH-FSH-S11. Ovaries from 29 day old rats transplanted to other prepubertal animals (23...

Induction of a pale, soft, exudative-like myopathy and sudden death in pigs by injection of anterior pituitary extract.

Yorkshire pigs, 60 days old, were used to determine whether pale, soft, exudative (PSE) muscle and the porcine stress syndrome (PSS) could be induced by prolonged injections of desiccated porcine pituitary powder (PP). Three barrows and three ovariectomized gilts, negative for PSS by the halothane and CPK test, were assigned to each of the following groups: (1) control saline; (2) 50 mg of PP and (3) 100 mg of PP in saline. Injections were given intramuscularly in split doses, daily for 8 weeks. At 4 weeks, the level of PP was doubled. At 4 weeks, one pig of group 3 died, and by 8 weeks one pig of group 2 and two other pigs of group 3 died after displaying typical signs of PSS. Animals were anesthesized before exsanguination, and carcasses were held at room temperature. At time 0 postmortem PP-injected pigs had lower (P<.05) longissimus muscle pH, higher (P<.05) carcass temperature, lower (P<.01) muscle glycogen, higher (P<.001) muscle lactate and lower (P<.001) muscle creatine phosphate than saline-injected (control) pigs; whereas, muscle G-6-P tended to be higher (P<.1) in the PP-injected animals than in the control pigs, and muscle adenosine 5'-triphosphate (ATP) levels were similar in all treatments. PP-injection increased the rate of longissimus muscle postmortem glycolysis as shown by more rapid pH decline and ATP depletion and more rapid lactate accumulation. Hunter L and transmission value (sarcoplasmic protein solubility) of longissimus muscle indicated that the PP-injected pigs also displayed poorer quality muscle than the control pigs. Blood levels of glucose and lactate throughout the treatment period indicated that PP increased the general metabolic rate. Blood creatine phosphokinase (CPK) was also elevated by PP between 4 and 6 weeks of treatment. On the basis of visual appraisal of the longissimus muscle, none of six pigs in group 1, three of five pigs in group 2 and one of three pigs in group 3 displayed PSE musculature. The results indicate that prolonged injections of pituitary extract induced a PSE-like myopathy and sudden death, preceded by symptoms of PSS.

Heterogeneity of rat luteinizing hormone revealed by radioimmunoassay and electrofocusing studies.

Radioimmunoassay of rat luteinizing hormone (LH) in the sera and the anterior pituitaries of normal and castrated males and proestrous females employing different batches of anti-LH sera gave the results suggesting heterogeneity of immunoreactive LH (IR-LH). Gel filtration analyses of anterior pituitary extract and pooled sera failed to give positive evidence for the heterogeneity. But when the extracts of normal rat anterior pituitaries were electrofocused and the fractions were assayed, the pituitary glycoprotein hormones, LH, FSH and TSH, were found to consist of several components with different isoelectric points. In electrofocusing patterns of IR-LH, distinct peaks were observed at pH 8.5 (component A), 8.8 (component B), 9.1 (component C) and 9.3-9.4 (component D), and less distinct peaks were observed at pH 7.9 (component A'), 9.6 (component E) and 9.8 (component F). There seemed to be some sexual difference in the relative amounts of these components. Orchiectomy caused marked changes in electrofocusing patterns of IR-LH, i.e., increase of components A', A, B, C and D. The blockade of LH release by estradiol benzoate and progesterone treatment in orchiectomized rats caused a marked increase of component A. At the proestrous LH surge, IR-LH components were evenly released. Radioimmunoassay of IR-LH components with multiple antisera indicated that these antisera had different affinities to these components.

Inconsistency of the tibia test for estimating growth hormone in crude pituitary extracts.

The complement fixation immunoassay (CFIA) was used for quantitating growth hormone (GH) in crude anterior pituitary extracts from rats subjected to thyroidectomy with or without cortisol and exogenous GH administration. The results obtained from this study were compared with pertinent bioassay (tibia test) results or correlated with the pituitary acidophil cell counts. Whereas it has been reported that pituitary GH levels are normal by the tibia test at 2 weeks after thyroidectomy, the highly specific CFIA method showed an actual 87% decrement which correlated well with the reduction in acidophilis. In addition, the apparently normal content of GH after cortisol administration to thyroidectomized rats, as measured by the tibia test, was contradictory to the very low acidophil population and to the marked reduction in pituitary GH content as measured by the CFIA. Furthermore, theoretical tibia responses illustrate the inconsistency of the bioassay in different experimental conditions. If, as previously suggested, the content of thyrotrophin (TSH) in the crude pituitary extracts renders the bioassay of GH a dubious procedure, then the superiority of immunoassay is obvious.

The control of time of oestrus and ovulation and the induction of superovulation in cattle

In an attempt to induce superovulation at predetermined times 140 mature parous cows and 27 heifers were treated with progesterone by intramuscular injection (40 mg/day for 17–30 days) or prostaglandin F2α (PG) by intrauterine injection on each of two consecutive days (500 µg/injection) between the 8th and the 14th day of the oestrous cycle. All animals were given pregnant mare serum gonadotrophin (PMSG) or an equine anterior pituitary extract (HAP) at one of three doses, and human chorionic gonadotrophin (HCG) at a standard dose of 1500 i.u. was given to two-thirds of the cows on either the second or third day after the final progesterone or first PG injection. Following treatment the animals were run with entire bulls and all were slaughtered within 7 days of the end of treatment. During treatment progesterone effectively inhibited ovulation, while PG caused luteolysis in the vast majority of animals. Ninety-eight cows and 23 heifers were in oestrus within 6 days of the final progesterone or first PG injection, and the majority (87%) within 2–4 days. All 27 heifers and 129 of the 140 cows ovulated within a few days after treatment, and there was no apparent difference in either time of ovulation or ovarian response between animals which did, and did not, exhibit oestrus. PMSG and HAP were equally effective in inducing superovulation, but in heifers HAP gave more unruptured large follicles than did PMSG. Overall, the ovulatory response of heifers was greater than that of cows. HCG, irrespective of when given, had no effect upon either incidence of oestrus or ovarian response. In cows and heifers which were in oestrus, the respective percentages of eggs shed that were recovered from the reproductive tract, and the percentages of recovered eggs that were fertilized, were 73 and 79, and 78 and 95. There was no effect of any factor other than HCG on either recovery or fertilization rates. In cows but not in heifers, HCG treatment at either time appeared to reduce recovery and fertilization rates. Where numbers of fertilized cow eggs are required, it is suggested that PMSG in conjunction with PG provides the most simple treatment. Further, it seems desirable to restrict treatment to heifers or young cows.

Regeneration of the forelimb in adult hypophysectomized Notophthalmus (Diemictylus) viridescens given embryonic or adult chicken anterior pituitary extract.

AbstractIn absence of the anterior pituitary gland, limb and tail regeneration does not occur in the adult newt Notophthalmus (Diemictylus) viridescens. Sixty‐five adult newts were included in this work which was designed to test to what degree a hypophysectomized newt can regenerate its forelimb when given daily injections of fresh adult or stored (−20° C)‐thawed embryonic or adult chicken anterior pituitary gland extract. Injections were begun seven days after hypophysectomy (to minimize further the titres of residual circulatory hormones) and concomitant amputation. The results are based on three experimental, one control and one sham series.In 40 out of 45 cases in Experiments C, D, and E, limb regeneration ensued for 25–30 days post‐amputation (until fixation). However, in the Experiment E newts, which were given embryonic chick pituitary extract, the regenerates were less advanced than those seen in the animals of Experiments C and D, given adult chicken pituitary extracts. Possible explanations for the differences in results are discussed.The results show that forelimb regeneration and survival in our hypophysectomized animals were due to extracts of embryonic or adult chicken anterior pituitary glands; presumably, these extracts contained the essential hormones. On the basis of our newt bio‐test system, 15–18 day chick embryo and adult chicken anterior pituitary extracts (hormones) are not species specific. Freezing and storage of extracts at −20° C for up to three months before thawing and use apparently does not appreciably affect their hormonal activity.

A re-examination of the tadpole in metamorphic stasis as a recipient in a bioassay for thyroid-stimulating hormone.

SUMMARY The tadpole, in metamorphic stasis, has been used as a bioassay recipient for thyroid-stimulating hormone (TSH). The validity of its use for this purpose, however, has not been tested critically, particularly with respect to the effect of other hormones, most notably, growth hormone (GH) and luteinizing hormone (LH). Growth hormone has been shown to influence thyroid function and LH is a common contaminant of pituitary TSH preparations. Tadpoles of Rana pipiens, arrested at a particular metamorphic stage, received various concentrations of LH alone, a combination of doses of GH and TSH, or GH and rat anterior pituitary extract. Growth hormone was ineffective in re-inducing metamorphosis in 78% of cases, whereas LH stimulated metamorphosis in the tadpoles.

FSH pleomorphism in the rat--regulation by gonadal steroids.

Molecular distribution profiles for follicle-stimulating hormone (FSH), luteinizirig hormone (LH), and thyroid-stimulating hormone (TSH) in anterior pituitary extracts from intact and gonadectomized rats of both sexes, androgen-treated orchidectomized rats, and estrogen-treated ovariectomized rats were obtained by exclusion chromatography. Hormonal activities of the extracts and of the eluted fractions were determined by radioimmunoassay. Two internal “markers”, serum albumin (SA) and ovalbumin (OA), were added to each aliquot of pituitary extract before it was applied to the column. The locations of the FSH, LH and TSH peaks were plotted in relation to those of these two standard proteins. The elution position of TSH was relatively constant; it invariably peaked just ahead of the OA marker, behind FSH and LH. Generally, LH also varied little in location, although the LH from female rats did tend to be retarded slightly. By contrast, there were distinct differences among the FSH peaks. FSH from males and ...

Effect of progesterone and oestrogen on the survival and development of fertilized ova in the ovariectomized ewe.

Two experiments were carried out to investigate the importance of progesterone and oestrogen for the survival and development of fertilized ova in ovariectomized ewes. In experiment 1 ewes were treated with progesterone (P) alone or progesterone and 17 p-oestradiol (P + E2) following ovariectomy on the second (day 2) or sixth day (day 6) after mating to vasectomized rams. Fertilized ova were transferred to the ewes either immediately after ovariectomy or on day 6 after ovariectomy on day 2. In experi-ment 2 ewes that had been given an equine anterior pituitary extract to induce multiple ovulation were ovariectomized on day 2 after mating to fertile rams, and then treated with P or P+E2. On day 18 the uteri of all ewes were flushed to recover embryos. Treatment with P commenced immediately after ovariectomy and E2 was given from day 21-to day 5, to ewes ovariectomized on day 2. Ovarian vein blood was collected just prior to ovariectomy and was assayed for its progesterone and oestrogen contents.

Bioassay of thyrotropin in chicks with simultaneous estimation of gonadotropin

Simultaneous determinations of thyrotropin and gonadotropin content of anterior pituitary extracts and anterior pituitary glands was made utilizing White Leghorn cockerels 12–24 hr after hatching. The uptake of 32P per milligram wet weight of thyroids or testes provided an excellent end point for estimation of hormone activity. A 5-hr treatment with hormone coupled with a 1-hr uptake of 0.3 μCi to 1.0 μCi of 32P proved to be an effective procedure. An exposure to 32P for as little as 15 min was adequate for thyroid responses, but this interval was not satisfactory for differential testes reactions. Excellent estimations of activity were obtained with milliunits of thyrotropin especially with a 3 × 3 factorial assay design. Marked differences were noted in thyrotropic and gonadotropic titers when National Institutes of Health thyrotropin (NIH-TSH) was compared with Nutritional Biochemicals Co. thyrotropin (NBC-TSH) and with Sigma Chemical Co. thyrotropin (Sigma-TSH). Pretreatment with thiourea improved the precision of the assay but responses to TSH were much lower. Assays for the thyrotropic activity of NIH-LH and NIH-FSH preparations demonstrated that appreciable TSH activity was present in the LH but not in the FSH. The relative amounts of thyrotropin present in the two gonadotropin preparations agreed closely with the estimations stated in the NIH specifications for the two hormones. Anterior pituitary glands of 28-day-old and 62-day-old cockerels contained equivalent amounts of thyrotropic activity per milligram of gland. The pituitaries of the younger birds, however, had a gonadotropin titer which was twice as great as that of the older cockerels.

Inhibitory effect of pituitary factor on phagocytosis and iodine metabolism in human leukocytes.

A pituitary factor decreasing iodine uptake and protein-bound iodine formation in human leukocytes was found in semi-purified preparations of TSH, LH, FSH, GH and in crude anterior pituitary extract. ACTH, insulin, serum gamma globulin, myoglobin and ceruloplasmin were ineffective in comparison to control samples incubated with serum albumin. TSH, in relation to dose (0.01–0.5 U/ml), significantly decreased the total uptake, formation, and release of iodocompounds from phagocytic leukocytes. The amount of iodocompounds released from resting or phagocytic leukocytes into the media was twice as high as the amount of organically bound iodine that remained in the cells. These data suggest that iodine-containing compounds are actively formed, metabolized, and released from human leukocytes. Because TSH also decreases engulfment of latex microparticles, the observed changes in iodine metabolism are probably secondary, due to the reduced phagocytosis. The presence of an unknown factor in the semipurified prepara...

Fertilization in ewes treated with progesterone and equine anterior pituitary extract.

In recent experiments in which large numbers of fertilized eggs were required on specific days, 30 Merino ewes experiencing regular oestrous cycles were treated with progesterone to control the time of oestrus and ovulation, and with an equine anterior pituitary extract (HAP) to induce multiple ovulation. Progesterone was administered daily (10 mg/day i.m.) beginning on the 8th to 12th day of the oestrous cycle and continuing for 6–8 days. HAP, prepared as described by Moore & Shelton (1964), was given as three equal, daily s.c. injections (24 mg./day) beginning on the day before the final progesterone injection and ending the day after the final injection of progesterone. The ewes were run with Merino rams and were inspected for oestrus at 12-hr. intervals after the conclusion of treatment. All 30 ewes were served after treatment and the time between the end of progesterone treatment varied from 36–84 hr. Laparotomies were performed

Further investigations on specificity and precision in the Igarashi-McCann FSH assay method.

The Igarashi-McCann method, reported in 1964 as a new biological method assaying FSH with MED corresponded to 2μg equivalents of NIH-FSH-Sl utilizing intact female mice of Swiss-Albino, would be appreciated to be indispensable for sensitive evaluation of FSH, in spite of having been suffered from poor precision which had been destined with the principle of uterine weight augmentation response. The optimum dose of HCG to be added was confirmed to be 0.075 iu through variance analysis of regression line and statistical investigation of linearity and stability in the dose response curve. The limitation and unification of beginning body weight of recipient were concluded to be inevitable not only for improvement of precision but also for qualitative unification in uterine weight response. Utilizing the unified recipient of 8 g, the stable and specific estimation of FSH in normal urine and plasma can be performed within such levels of maximum permitted contaminating dose (MPCD) as 2.0μg equiv. of NTH-LH-S13, 2.5μg equiv. of NIH-GH-S7, 1.0μg equiv. of NIH-GH-HS1082B, 400 mu equiv. of NIH-P-S7, 200mμ equiv. of Armour-ACTH, 200 mu equiv. of NIH-TSH-S5, and 0.005 μg of Estradiol with 0.075 iu of HCG, after referring reported contents of these hormones in pituitary, in plasma, and in urine. In conclusion, the results of Igarashi-McCann assay are specific for FSH, if less than 14.4 μg of rat anterior pituitary extract or less than 2.0ml of rat or human plasma, or 1.0 hr and 0.2 hr equiv. or less of non-pregnant human urinary extract except of acromegaly is injected into one mouse, respectively.