bluuebunny/crossref_metadata_2025 huggingface dataset:
THE ASSESSMENT OF ADRENAL-CORTICAL FUNCTION IN PATIENTS RECEIVING CORTICOTROPHIN THERAPY
Authors: Levell M. J. , Stitch S. R. , Noronha M. , Vas C.
Published: December 2008

Plasma cortisol and urinary 17-hydroxycorticosteroids and cortisol secretion rates have been measured on patients with multiple sclerosis taking part in a therapeutic trial of corticotrophin. Patients who had not received corticotrophin showed no evidence of abnormal adrenal function, although the combination of normal levels of plasma cortisol with somewhat low concentrations of urinary cortisol metabolites suggested an abnormal steroid metabolism. Patients who had been treated continuously with corticotrophin (for at least 1 year) showed elevated plasma cortisol concentrations and an even greater disparity between plasma and urine concentrations than the untreated controls. Thus, the urinary steroid measurements did not give a useful measure of the degree of adrenal response and should not be used to monitor prolonged corticotrophin therapy.
BESTIMMUNG VON TESTOSTERON IM PERIPHEREN BLUT VON SCHWEIN UND RIND MIT EINER DOPPELISOTOPEN-DERIVAT-VERDÜNNUNGSMETHODE
Authors: Hoffmann B. , Claus R. , Karg H.
Published: December 2008

Using a double-isotope-dilution derivative method combined with a cristallisation technique testosterone was measured in pig and bovine peripheral blood. In the male pig the values varied from 0.94 to 1.61 μg per 100 ml plasma and were 7- resp. 11-times higher than in castrated and female animals. Values increased with age. The concentration measured in a bull was 0.59 μg per 100 ml plasma.
METABOLISM OF OESTRADIOL-17α-3H AND OESTRADIOL-16β-3H IN PREGNANT WOMEN
Authors: de la Torre B. , Wiqvist N. , Diczfalusy E.
Published: December 2008

The metabolism of oestradiol-16-14C,17α-3H and oestriol-16-14C,16β-3H was studied following the intravenous administration of each compound to two women at midgestation. In addition, the foeto-placental metabolism of oestriol-16-14C,17α-3H was investigated in a subject following the administration of the tracer into the umbilical circulation at laparotomy. The 3H/14C ratios of urinary, foetal and placental oestriol conjugates were estimated following reverse isotope dilution of the oestriol moiety and its triacetate.

The 3H/14C-ratios of the oestriol-16-glucosiduronate (OE3-16GL) and oestriol-3-sulphate,16glucosiduronate (OE3-3S,16GL) fractions isolated from all urine specimens, as well as those of unconjugated oestriol (OE3), oestriol-3-sulphate (OE3-3S) and OE3-16GL isolated from the foeto-placental unit did not differ from each other and agreed with that of the administered material. Also, the 3H/14C-ratio of the oestriol-3-glucosiduronate (OE3-3GL) isolated from the urine excreted during the first 24 h, agreed with those of the other conjugates. However, a statistically highly significant drop was found in the 3H/14C-ratio of the OE3-3GL isolated from the second and third 24 h urine collections. Relatively more 3H was lost from oestriol-16β-3H, than from oestriol-17α-3H.

It is concluded that OE3-3GL underwent a limited oxidation both at C-atoms 16 and 17. This oxidation seems to take place in the gastrointestinal tract and might be due to the activity of microorganisms.

It is suggested that the same steroid molecule present in the form of various conjugates may be metabolized differently.
EFFECT OF 9:10-DIMETHYL-1:2-BENZANTHRACENE ON THE MOUSE OVARY
Authors: Krarup Torben
Published: December 2008

Administration of the carcinogenic hydrocarbon 9:10-dimethyl-1:2-benzanthracene (DMBA) in mice causes destruction of the oocytes in the ovaries resulting in a premature depletion of the germ cells. Subsequently ovarian tumours develop.

It was investigated to what extent fertility is influenced during the process of oocyte disappearance in immature or mature mice treated with DMBA. The first parturition was not delayed and as long as the treated mice remained fertile, litters were born at normal intervals. However, the fertility span was considerably shortened due to the premature depletion of oocytes, but in contrast to the control mice, young were born to the DMBA-treated mice until no or very few oocytes remained. As a consequence of the shortened fertility span, the experimental mice gave birth to fewer litters than the control mice. The litter sizes were also smaller; the possible reason for this reduction in size is discussed in relation to the DMBA-induced reduction in the pool of small oocytes.

One year after DMBA-ingestion a number of mice had developed ovarian tumours. Using the time of birth of the last litter as an indicator of the time at which the ovaries were depleted of oocytes it is shown that the earlier the germ cells disappeared, the earlier tumours developed.
KOMBINIERTE ELEKTRONENMIKROSKOPISCHE UND BIOCHEMISCHE UNTERSUCHUNGEN ÜBER DEN EINFLUSS VON TESTOSTERON AUF DAS VESIKULARDRÜSENEPITHEL JUNGER RATTEN
Authors: Dahnke Hans-Gerhard , Mosebach Karl-Oskar
Published: December 2008

The effect of testosterone on the morphology of the secretory cells in the seminal vesicles of 13 and 30 days old rats was studied by means of electron microscopy. After five administrations of the hormone (5 × 100 μg testosterone) to 13 days old animals there was an increase of free ribosomes, morphological changes in the Golgi complex and a marked enlargement of the intercellular spaces. No significant morphological changes could be demonstrated in rats aged 30 days.

In addition the effect of testosterone on the uptake of [2-14C]thymidine by the seminal vesicles of 13 days old rats was investigated. This uptake only slightly enhanced (about 30%). It was remarkable however, that an almost identical uptake of thymidine occurred in all animals treated with the hormone, whereas the uptake was quite different in the controls. Whether testosterone was administered once or five times did not affect the results.

The morphological and biochemical results suggest that the action of testosterone on the seminal vesicles of very young rats involves effects other than those seen in older animals.
EFFECT OF THERMAL INJURY ON EXCRETION OF HYDROXYPROLINE IN RATS, TREATED OR NOT WITH AN ANABOLIC ANDROGEN
Authors: Kowalewski K. , Heron F.
Published: December 2008

Urinary excretion of hydroxyproline, considered as an index of collagen metabolism, was determined prior to, and during, a period of healing of thermally injured rats' skin. Standard deep dermal burns (about 12% of the total skin surface) were produced in rats. Animals were left untreated or received, during the healing period, an anabolic steroid 17β-hydroxy-17α-methylandrostano [3,2-c] pyrazole (Winstrol®), Urine was collected prior to injury and after burns. Hydroxyproline was determined and expressed in μg per 24 h per 100 g of rats' body weight. Thermal injury affected significantly hydroxyproline excretion: progressive increase of this component of collagen was observed, with peak value in third week after burns. This was considered due mostly to the proteolysis of irreversibly denaturated collagen. However, some excreted hydroxyproline might represent the product of newly formed collagen of healing tissue. Anabolic steroid used in this study produced an increase in hydroxyproline excretion as compared with untreated controls, but this effect was apparent only during the third week after injury. Hydroxyproline excretion test may be of value following thermal injury of the skin. Experimental results in this respect, found in rats, are similar to those found previously in humans.
THE TIBIAL GROWING CARTILAGE BIOPSY IN THE STUDY OF GROWTH DISTURBANCES
Authors: Stănescu V. , Bona C. , Ionescu V.
Published: December 2008

51 biopsies of tibial growing cartilage were performed in children with and without growth disturbances. The biopsy procedure is described.

The clinical and roentgenological follow up did not reveal any untoward effects due to the operation. The histological and cyto-enzymological methods used in the study of the biopsies as well as the main results obtained are presented and discussed. In a few cases the effect of a growth stimulating treatment could be studied by means of a repeated biopsy from the opposite leg.
DETERMINATION DE LA TRANSCORTINE PLASMATIQUE AU COURS DE DIVERSES ENDOCRINOPATHIES
Authors: Burger A. , Laudat P. , Bricaire H. , Pique L.
Published: December 2008

The transcortin capacity for cortisol (CBC, μg cortisol/100 ml) has been determined by gelfiltration using [1,2-3H]cortisol (De Moor et al. 1962; Doe et al. 1964). There was always 2 to 3% unspecific binding to CBC free plasma (heated to 60°C for 30 minutes) and to pure albumin. The protein binding found in unaltered plasma had to be corrected by subtracting this unspecific radioactivity to calculate the CBC, which had a normal range of 24 ± 8 μg cortisol/100 ml plasma (2 sd, n = 35). Eight states of hyperoestrogenism, 5 hypopituitarisms, 8 acromegalies, 3 congenital adrenal hyperplasias, 2 hirsutisms and 12 Cushing syndromes were tested. As a group only the CBC in hyperoestrogenism was high and outside the normal range. The CBC in two cases of Cushing syndrome and in two cases of acromegaly were low. The CBC has been followed before and under treatment with 2,2(4-chlorophenyl,2-chlorphenyl)1,1-dichlorethane (op'DDD). No significant changes were seen.
EFFECTS OF AMINOGLUTETHIMIDE IN A CASE OF ECTOPIC ACTH SYNDROME
Authors: McMillan Mary , Maisey M. N.
Published: December 2008

A male patients with the ectopic ACTH syndrome due to bronchial carcinoma gained a partial remission of symptoms when treated with aminoglutethimide. The drug may have caused a haemolytic anaemia. A fall in plasma 11-hydroxy-corticosteroids was noted. There was a fall in urinary 17-hydroxy-corticosteroids, 17-ketosteroids and oestrogens with a rise in the aetiocholanolone/androsterone ratio, and reversal of the testosterone glucosiduronate/epitestosterone glucosiduronate ratio.
CORTISOL RELEASE, DISTRIBUTION AND METABOLISM IN INDUCED OESTROGENIC DEFICIENCY
Authors: Jansen Jens Aas , Schou Jens , Singh H.
Published: December 2008

The biological half-life of exogenous [4-14C] cortisol in guinea pigs is approximately 105 min. After ovariectomy there is a tendency to a prolonged half-life (delayed metabolism) of cortisol. The plasma cortisol in guinea pigs is 314 ± 79 ng/ml with an ultrafiltrable fraction of 38 ± 15 ng/ml. Skin cortisol is 44 ± 12 ng/g tissue. After ovariectomy the plasma cortisol is decreased (195 ± 36 ng/ml) and there is a higher relative diffusible fraction, while the skin cortisol is decreased (23 ± 4 ng/g). During stress release there may be a parallel release of a cortisolbinding non-ultrafiltrable component or an elimination of a protein bound not fluorescent or separated steroid. The tissue cortisol is higher than would be expected from the values of diffusible cortisol in plasma. The low concentration of ultrafiltrable cortisol may explain the reduced rate of cortisol metabolism.
EPIDERMOID CYSTS IN THE THYROID GLANDS OF MALE RATS TREATED NEONATALLY WITH OESTROGEN
Authors: Arai Y. , Maeda T. , Masuda S.
Published: December 2008

Epidermoid cysts developed most frequently in the thyroid gland of male rats which had been castrated on the day of birth and injected with oestradiol benzoate (OB) for 30 successive days from birth. The incidence of epidermoid cysts was 90% in this group. The daily doses of OB were 1 μg during the first 10 days, 2 μg during the next 10 days, and 4 μg from days 21 to 30, respectively. These metaplastic lesions were not reversible after cessation of OB treatment. When OB was injected into intact male rats for the first 30 days of life, the incidence was decreased to 30%. In contrast, there was no manifestation of epidermoid cyst formation in the glands of control rats such as in intact male rats which had been treated with OB from days 31 to 60, or neonatally castrated and intact male rats, both given oil only.
THE OVULATORY EFFECTS OF FOLLICLE STIMULATING HORMONE TREATED WITH CHYMOTRYPSIN IN CHLORPROMAZINE BLOCKED RATS
Authors: Harrington F. E. , Bex F. J. , Elton R. L. , Roach J. B.
Published: December 2008

Adult prooestrous rats were injected subcutaneously with chlorpromazine prior to the »critical period« (2:00 p. m.–4:00 p.m.) to prevent ovulation. Midway through the »critical period« (3:00 p. m.) these blocked females were injected intravenously with varying doses of follicle stimulating hormone which had been maintained overnight at room temperature or incubated with chymotrypsin overnight in a waterbath maintained between 35–36°C. Results showed that the amount of NIHFSH-S7 calculated to cause ovulation was nearly identical for enzyme exposed (15.49 μg) and unexposed (15.36 μg) FSH.

The Parlow assay was used to verify that the enzyme specifically digested the LH contaminant of FSH. Significant depletions of ascorbic acid were obtained only with NIHFSH-S7 samples that were unexposed to the enzyme. Overnight incubation of FSH with chymotrypsin completely prevented depletion of ovarian ascorbic acid, verifying that the enzyme digested the LH contaminant present in NIHFSH-S7. These data further support our concept that follicle stimulating hormone by itself induces ovulation in CPZ blocked rats.
SPONTANEOUS VARIATIONS IN SERUM GROWTH HORMONE LEVELS
Authors: Buckler J. M. H.
Published: December 2008

Serum growth hormone (HGH) levels have been estimated in hourly samples for 24 hours in 2 women and 1 man. The highest levels in all subjects were found in association with the onset of sleep at whatever time that occurred. Other spontaneous peaks were found particularly in 1 woman. The rise in serum HGH levels in women following the stimuli of exercise or Bovril was greater or more rapid when these stimuli were applied mid-morning compared with early in the morning. Serum HGH levels at the same times on different days showed a great variation in the mid-morning levels from day to day in a female subject. No such variation was found in a man, nor in the female subject in the early morning.
INTERRELATIONSHIP BETWEEN CALCITONIN AND PARATHYROID HORMONE IN INTACT, THYROIDECTOMIZED, PARATHYROIDECTOMIZED, AND THYROPARATHYROIDECTOMIZED RATS
Authors: Sørensen O. Helmer
Published: December 2008

Calcitonin is capable of completely suppressing the hypercalcaemic effect of parathyroid hormone but not irreversibly. The parathyroid hormone extract used was the commercial preparation Para-Thor-Mone® (Lilly). When injected simultaneously, the parathyroid hormone had no influence on the hypocalcaemia induced by calcitonin one hour after the injection. The parathyroid hormone produced a hypercalcaemia of up to 42 h duration in rats without thyroids. If the thyroid gland was present the serum calcium did not exceed 5.5 mEq./l, probably because of an increased secretion of calcitonin. A transitory hypercalcaemia followed by a hypocalcaemia was seen in intact rats after an intraperitoneal calcium load. The hypocalcaemia could be explained by the counteracting secretion of calcitonin.
EXTRA-THYROIDAL EFFECT OF THYROTROPHIN ON SERUM THYROXINE
Authors: Sisson James C.
Published: December 2008

Injections of bovine thyrotrophin (TSH) into thyroidectomized guinea pigs which were receiving maintenance doses of thyroxine (T4) reduced the PBI levels. The phenomenon was demonstrated only with relatively large quantities (0.8-6 U per day) of TSH, but the response was dosedependent. The quality to affect the PBI remained associated with the thyroid-stimulating activity of TSH when partial purification of the trophic hormone was achieved.

Multiple effects of TSH on T4 economy were found in these animals. Diminished binding affinity for T4 by serum proteins and enhanced deiodination of T4 by hepatic tissue could be demonstrated. The depression of the PBI by TSH appeared to be related to a redistribution of T4 which may, in part, have resulted from changes in the serum binding proteins. A relationship of increased hepatic deiodination of T4 to the decrement in PBI could not be established with certainty.

In doses that could be reasonably administered, bovine TSH had no effect on the thyroid hormone economy in an athyreotic man.
KINETICS OF DOUBLE-ANTIBODY RADIOIMMUNOASSAY SYSTEMS FOR HUMAN GONADOTROPHINS
Authors: Borth Rudi
Published: December 2008

Incubation time curves were investigated in double-antibody radioimmunoassay systems. It was confirmed that the first reaction is not always halted on addition of the second antiserum; depending on the condition of the system at the time of adding the second antiserum, the first reaction may continue slowly, come to a standstill, or be reversed. A kinetic interpretation is discussed. Under the experimental conditions employed, the distortion of incubation time curves produced by disregarding the observed phenomena was found to be slight.
NORMAL VALUES OF SERUM PROTEIN BOUND IODINE IN HEALTHY CHILDREN AGED BETWEEN 4 AND 13 YEARS; BIOCHEMICAL EVIDENCE FOR THE EXISTENCE OF THE JUVENILE OR MID-GROWTH SPURT
Authors: van Stekelenburg G. J. , Stoop J. W. , van der Stam P. J.
Published: December 2008

Protein bound iodine (PBI) was determined in 379 serum specimens of boys and girls, aged between 4 and 13 years. From this investigation it can be seen that: (a) there is no significant difference in the PBI concentration between boys and girls of the same age group, (b) in general the PBI concentration decreases with increasing age (1.4 μ/l/year), (c) both for boys and for girls there is a significant increase in the PBI concentration from the sixth year to the seventh year of age, a result which coincides with the juvenile or mid-growth spurt, and (d) the normal range, calculated from all data, amounts to 40.7–71.1 μg/l (mean value ± 2x standard deviation).
EFFECT OF SYNTHETIC HUMAN 1,39-CORTICOTROPHIN ON BLOOD GLUCOSE LEVEL IN MICE
Authors: László F. A. , Szijj I. , Durszt F. , Kovács K.
Published: December 2008

The hypoglycaemic action of synthetic human 1,39-corticotrophin was determined in mice and its effectiveness compared with highly purified porcine corticotrophin.

Synthetic human 1,39-corticotrophin (0.01 mg = 1.0 IU) and porcine corticotrophin (1.0 IU) induced a transient hypoglycaemia. After the administration of the porcine corticotrophin the decrease in the blood glucose concentration was somewhat more marked and prolonged. Adrenocortical activity was not necessary for the development of hypoglycaemia. In adrenalectomized mice, dexamethasone substitution by increasing the initial blood glucose content, made the effect more pronounced. Pretreatment with corticotrophin reduced the extent of the alloxan-induced transitory hyperglycaemia. On the other hand, it did not influence the high blood glucose values in manifest alloxan diabetic animals.

It is possible that corticotrophin induces hypoglycaemia through insulin release. This is an extra-adrenal effect of corticotrophin as it is also observed in adrenalectomized mice.
PARTIAL SEPARATION OF A 3α-KETOSTEROID OXIDOREDUCTASE AND AN ANDROGEN BINDING SUBSTANCE PRESENT IN RAT VENTRAL PROSTATE CYTOPLASM
Authors: Unhjem Olav
Published: December 2008

The presence of a 3α-ketosteroid oxidoreductase in the rat ventral prostate cytoplasm has been demonstrated. The enzyme was confined to the 105 000 × g supernatant fraction with very little activity in the mitochondrial-microsomal fraction. Addition of NADPH2 was necessary for the enzymatic reaction to proceed. The enzymatic activity was lost when supernatant fractions were dialysed against 0.1 m Tris-HCl buffer, pH 7.4 or when chromatographed by gel filtration using the same buffer. When either L-cysteine, reduced glutathione, 2-mercaptoethanol or EDTA was added to the buffer, the enzymatic activity was preserved. The molecular weight of the enzyme was calculated to be 40 000–50 000. 3H-5α-dihydrotestosterone associates with macromolecules in the 105 000 × g supernatant fraction which are of two molecular sizes (60 000–70 000 and ∼ 200 000). These complexes could not be demonstrated by gel filtration using buffers containing either reduced glutathione, 2-mercaptoethanol or EDTA. By using L-cysteine, the small molecular complex was preserved.
EFFECTS OF OESTROGEN ON SERUM LEVELS OF LH AND FSH
Authors: Nillius Sven Johan , Wide Leif
Published: December 2008

The acute effects on the serum levels of LH and FSH after the administration of some oestrogen preparations were investigated in oligoamenorrhoeic women, in postmenopausal women and in men. Immunoreactive LH and immunoreactive FSH in the serum were assayed by a radioimmunosorbent technique.

The diurnal rhythm in the secretion of the gonadotrophins was first studied. No significant circadian variation in either LH or FSH levels in the serum of hospitalized women or healthy men was found with this radioimmunoassay procedure.

The intravenous or intramuscular administration of 17β-oestradiol in doses from 5 μg to 5 mg resulted in a decrease in the serum levels of LH in 14 of 15 women studied and in the serum levels of FSH in 10 of 12 women. This can be explained by a negative feedback effect on the hypothalamic-pituitary system during the period of observation. The decrease in the serum gonadotrophin levels was as a rule more pronounced for LH than for FSH. No positive feedback effect on the hypothalamic-pituitary system following oestrogen treatment was observed. In men insignificant and variable changes in the serum gonadotrophin levels were found. This could possibly be explained by the low oestrogen doses used.
CIRCADIAN VARIATIONS IN ADRENAL SECRETION OF LEMMINGS, VOLES AND MICE
Authors: Andrews Richard V.
Published: December 2008

Daily variations in adrenal secretion by glands from arctic rodents were measured in vitro. Serial-sacrifice, short-term incubation studies yield similar results to those data obtained through organ-culture methods. Adrenal secretory rates display some desynchronization from locomotor activity during the summer solstice, although circadian variation persists during all seasons of the arctic year.
IN VITRO OESTROGEN BIOSYNTHESIS IN FOETAL ORGANS OF MIDPREGNANCY
Authors: Wu Chung-Hsiu , Archer David F. , Flickinger George L. , Touchstone Joseph C.
Published: December 2008

Foetal organs; namely adrenal, gastrointestinal tract, kidney, liver, lung, skin and testis of midpregnancy were incubated individually with radio-active dehydroepiandrosterone (DHA) and the free and conjugated oestrogens were analyzed. Free oestriol was isolated in the gastrointestinal tract, liver and skin incubates. Oestrone and oestriol glucuronides were found in the gastrointestinal tract and liver incubates respectively. In the gastrointestinal tract incubate oestrone sulphate was found in the highest concentration while sulphates of oestradiol and oestriol were shown in the liver incubate. Conversion rates were small, ranging from 0.009% to 0.121%. These studies suggest that although certain foetal organs are able to aromatize steroids, their contribution to the high levels of oestrogen produced by the foeto-placental unit may be negligible.
WEIGHT LOSS IN OBESE-HYPERGLYCAEMIC AND NORMAL MICE FOLLOWING TRANSAURICULAR HYPOPHYSECTOMY BY A MODIFIED TECHNIQUE
Authors: Herbai Georg
Published: December 2008

Hypophysectomy was performed by a modified transauricular procedure in normal (N. M. R. I.-strain), obese-hyperglycaemic mice and their lean litter mates. In order to obtain a high percentage of operation success the dimensions of the cannula were carefully adapted to the cranium sizes of the mice of both strains and the cannula fitted with a stop-disc. Abrupt post-operative growth arrest was shown to be a rapid in vivo indicator of the completeness of the operation. In obese-hyperglycaemic mice hypophysectomy caused a rapid decrease of the body weights by disappearance of large amounts of fat. Fourteen days after the operation successfully hypophysectomized obese mice became apparently normal. It is suggested that the pituitary plays a role in the maintenance of the syndrome.
EFFECTS OF PORCINE CALCITONIN ON SOME URINE ELECTROLYTES IN THE RAT
Authors: Aldred J. P. , Stubbs R. K. , Hermann W. R. , Zeedyk R. A. , Bastian J. W.
Published: December 2008

Bidaily (b. i. d.) subcutaneous administration of porcine calcitonin (CT) to intact rats resulted in dose-related increases in urinary excretion of water, calcium, phosphorus and magnesium. The effects required several days of treatment to reach maximum levels and persisted for several days after treatment ceased.

Although hypocalcaemia occurred immediately following CT injections in chronically treated rats, animals treated for 8 days were significantly hypercalcaemic 18 hours later.

Bidaily administration of CT (9 a. m. and 3 p. m.) induced a distinct 24 hour periodicity in urinary excretion patterns of calcium, magnesium and urine volume, with maximal excretion at night. In the case of magnesium, this represented a direct reversal of the diurnal excretion pattern seen in control rats.

Urine volume and urine calcium were increased in parathyroidectomized (PTX) and thyroparathyroidectomized (TPTX) rats treated daily with CT. Surprisingly, the initial low blood calcium levels of these rats were elevated toward normal with daily CT treatment and returned to pretreatment levels following cessation of CT injections.

PTX rats treated with parathyroid extract (PTE) exhibited hypercalcaemia and hypercalciuria. Hypercalcaemia was only partially prevented by simultaneous treatment with high doses of CT and hypercalciuria was essentially unaltered.
A GRAPHICAL METHOD FOR THE COMPONENT ANALYSIS OF BINARY HORMONE MIXTURES: DISSIMILARITY OF PITUITARY AND URINARY GONADOTROPHINS IN RADIOIMMUNOASSAY
Authors: Borth Rudi
Published: December 2008

A graphical method using Loewe's isobols is described for the characterization of assay systems in which two agents produce a response, and for the component analysis of binary mixtures. The method is illustrated by describing the response surface of two radioimmunoassay systems for human gonadotrophins (tracer-FSH – anti-FSH and tracer-LH – anti-HCG) in terms of an FSH and an LH standard of pituitary origin. An attempt to assay the immunoreactivity of the second International Reference Preparation of Human Menopausal Gonadotrophins (HMG-IRP-2) produced strong evidence of dissimilarity, supporting the conclusion that HMG-IRP-2 is not a suitable reference standard for the radioimmunoassay of pituitary materials. The need for separate international research standards of human FSH and ICSH (LH) is emphasized.
PITUITARY LH CONTENT DURING DELAYED IMPLANTATION INDUCED BY DIFFERENT TREATMENTS IN RATS
Authors: Labhsetwar Anant P.
Published: December 2008

The capacity of oestrogens to induce nidation during delayed implantation and the known dependence of oestrogen secretion on LH prompted a detailed study of the pituitary LH content of rats in which implantation of blastocysts was delayed by various procedures. Implantation was delayed by Provera (2 mg), reserpine (200 μg), oral dosing with the antioestrogen I.C.I. 46,474 (40 μg) or clomiphene (100 μg), ovariectomy followed by progesterone treatment and finally by permitting mothers mated postpartum to suckle a large litter (10 pups). The pituitary LH stores (OAAD method) of such animals were compared with those of rats killed in prooestrus and oestrus and on day 9 of pseudopregnancy, pregnancy or lactation.

Hypophyseal LH levels were very low during delayed implantation induced by Provera or suckling and very high during that induced by ovariectomy followed by progesterone treatment compared with those in prooestrous, pseudopregnant or pregnant controls. By contrast, no significant change in LH stores was observed when nidation was delayed by single injections of I.C.I. 46,474 or clomiphene. Delayed implantation could not be induced in intact rats with chlormadinone or norgestrel even when given in large doses (2–8 mg/day). It is postulated that to delay implantation in the presence of the ovaries, a centrally acting agent must be capable of decreasing synthesis of LH in the pituitary.
THE EFFECT ON BONE OF LONG-TERM TREATMENT OF INTACT, PARATHYROIDECTOMIZED AND THYROPARATHYROIDECTOMIZED RATS WITH CALCITONIN
Authors: Hindberg Inge , Sørensen O. Helmer
Published: December 2008

Intact, parathyroidectomized, and thyroparathyroidectomized young, male Wistar rats were treated with sc injections of 80 MRC mU of calcitonin twice daily for three weeks and compared with similar controls receiving the vehicle for calcitonin. The serum calcium responses to single injections of calcitonin on the 10th and the 21st experimental days were found to be identical in the calcitonin-treated animals and their corresponding controls.

The results of calcium and magnesium determinations in the sternal manubria and in the left clavicles showed no evidence of an increased mineral deposition in the bones of calcitonin-treated rats, nor did isotope studies with 45Ca or survey X-rays and microdensitometer scannings of two caudal vertebrae.

The results in the intact rats can be explained by an increased parathyroid function as a counteraction to calcitonin. Rats with no parathyroid glands will have a greatly reduced bone resorption and will therefore show a diminished response to calcitonin. This can explain the observation that mineral deposition in bone is not increased in thyroparathyroidectomized and parathyroidectomized rats after long-term treatment.
COMPARATIVE BIOLOGICAL EFFECTS OF 125I AND 131I ON THE RAT THYROID
Authors: Vickery Austin L. , Williams E. Dillwyn
Published: December 2008

125I emits no β rays but part of its soft γ radiation is transformed into low energy electrons. Considering the short tissue path length (2-5 μm) of these electrons, it was postulated that its radiation effect on the thyroid might be different from that of 131I (path length 2000 μm). An experiment was designed to compare the biological effect of several dose levels of radiation from 125I on the rat thyroid gland with that from 131I. The effect of varying doses of each isotope on goitrogenesis (methylthiouracil induced) and on thyroid/serum radioiodide concentration (T/S) ratio was measured in animals radiated while on either a normal or a low iodide diet.

In relation to 131I, the effect of 125I on goitrogenesis generally was much less than its effect on function. The low iodide diet group given 125I, surprisingly, showed markedly less inhibition of goitrogenesis than the normal diet group, an opposite effect to that attained with 131I. We suggest that the observed differences are due to the gradient of energy absorption in the follicular cell from irradiation from 125I stored in the colloid. The apical cytoplasm would absorb much more energy than the basal cytoplasm and the nucleus, in contrast to the uniform absorption of energy from 131I across the cell. The comparatively lower effect of 125I on goitrogenesis, and the paradoxical reduction in its effect in the low iodide diet groups, may be due to the low energy absorption by the nucleus, especially when, in a taller cell, the nucleus is farther from the colloid.
ANALYSIS OF RADIOIMMUNOASSAY BY DISC ELECTROPHORESIS AND SUCROSE GRADIENT CENTRIFUGATION
Authors: Valenta Lubomir , Aubert Michel L.
Published: December 2008

Radioiodine-labelled synthetic adrenocorticotrophic hormone (ACTH), human growth hormone (HGH), human chorionic somato-mammotrophin (HCS), and human (HTSH) and bovine (BTSH) thyroid stimulating hormones were studied by disc-electrophoresis and sucrose gradient centrifugation before and after incubation with corresponding antisera. All antisera contained 7 S antibodies. After incubation, soluble antigenantibody complexes besides a small amount of precipitate were observed in the incubation mixture, characteristic of each hormone. The complexes migrated like gamma globulins or more slowly on disc-electrophoresis. and on sucrose gradient centrifugation showed patterns dependent on the time of incubation. Light 7 or 9 S, or < 12 S complexes occurred mostly after incubation for several minutes (up to 30 min) before analysis. When incubation was prolonged to 24 h and more, these relatively light complexes disappeared or diminished in favour of heavier soluble or precipitating complexes. Reproducibly obtainable sedimentation patterns of the soluble complexes suggested some definite recombination of antigen molecules with 7 S antibodies. The complexes did not occur on incubation with other sera than an antiserum to a given hormone. They were not influenced by EDTA. Displacement of the radioactivity of the complexes into the free hormone peak was obtained by addition of a non-labelled hormone identical with the labelled one. Sucrose gradient centrifugation and disc-electrophoresis are recommended for the study of immunoreaction of diluted materials and for a separate analysis of different steps of the radioimmunoassay.

Radioimmunoassay was introduced for the measurement of protein hormones by Yalow & Berson (1960). The method, described originally for insulin, was later adapted to the detection of a number of protein and polypeptide hormones. On incubation of the hormone with its antiserum, a soluble antigenantibody complex is formed, which is separated from an excess of the free hormone by various methods, e. g. chromatoelectrophoresis, precipitation with a second antibody, adsorption on a solid phase etc. (Hunter 1967).

Sucrose gradient centrifugation and disc-electrophoresis were occasionally used to follow some isolated aspect of radioimmunoassay (Fitschen 1965; Monjardino et al. 1968).

We are demonstrating that these methods made it possible to analyze the radioimmunoassay step by step and thus may be useful for practical purposes as well as in a study of the immunoreaction of diluted materials.
ELIMINATION AND METABOLISM OF 6-CHLORO-17α-HYDROXY-16-METHYLENE-4,6-PREGNADIENE-3,20-DIONE ACETATE AND ITS ANALOGUES IN RATS
Authors: Čekan Z. , Horešovský O.
Published: December 2008

The recovery of radioactivity in the urine, faeces and carcasses following single oral doses of [4-14C] progesterone, 17α-hydroxyprogesterone [2-14C] acetate (14C-AP), 16-methylene-6-dehydro-17α-hydroxyprogesterone [2-14C] acetate (14C-MAP), 6-bromo-16-methylene-6-dehydro17α-hydroxyprogesterone [2-14C] acetate (14C-BrMAP), 6-[36Cl] chloro-16-methylene-6-dehydro-17α-hydroxyprogesterone acetate (36Cl-ClMAP) and 6-chloro-16-methylene-6-dehydro-17α-hydroxyprogesterone [2-14C]-acetate (14C-ClMAP), respectively, was followed during 4–11 days. The total recovery was almost 100 per cent in all the compounds bearing the 17α-[14C] acetoxy group (AP, MAP, ClMAP, BrMAP). Comparison of 14C-ClMAP and 36Cl-ClMAP revealed that about 25 per cent of the administered radioactivity is eliminated in the urine in the form of desacetylated metabolites. However, most of the liberated acetic acid is not exspired, but is probably bound in an unknown form in the organism and excreted in the faeces.

The half-life of 36Cl-ClMAP is longer than that of the steroids labelled with 14C in the 17α-acetoxy group (14C-ClMAP, 14C-BrMAP, 14C-MAP, 14C-AP). This fact can be accounted for by a prolonged elimination of desacetylated metabolites.

The elimination in the urine of individual compounds was studied with ClMAP, AP, and MAP, the former two compounds being analyzed by double isotope analysis, the last one by single-label technique. The half-life of the individual compounds is shorter than that of the total radioactivity. This indicates that the individual compounds are eliminated faster than most of their metabolites. From the amount of unchanged steroids in the urine (related to the total radioactivity) it is apparent that ClMAP is metabolized less than AP, with MAP probably in an intermediate position. This correlates well with the biological activity of the compounds studied.

A preliminary study showed that following the administration of 14C-ClMAP a major portion of the radioactivity was found in both the urine and faeces in a non-conjugated, mostly neutral form. The majority of the neutral metabolites consisted of highly polar compounds. None of the metabolites separated by thin-layer chromatography, however, could be identified.
EFFECT OF AGE, SEX, STARVATION, HYPOPHYSECTOMY AND GROWTH HORMONE FROM SEVERAL SPECIES ON THE INORGANIC SULPHATE POOL AND ON THE INCORPORATION IN VIVO OF SULPHATE INTO MOUSE COSTAL CARTILAGE
Authors: Herbai Georg
Published: December 2008

The influence of several conditions on sulphate incorporation into costal cartilage was investigated in mice of the N.M.R.I.-strain and in dwarfmice (dw/dw) by a new double isotope technique. The method permits estimation of sulphate incorporation into costal cartilage in vivo after correction for the size of the exchangeable inorganic sulphate pool of the individual animal.

Detoxication to phenyl sulphate of injected phenol, following an injection of 3H-phenol and 35S-sulphate into the mice, allows determination of the sulphate pool. The incorporation rate of sulphate is expressed in ng, and includes all SO4 which had been incorporated in vivo into 5 pairs of cartilaginous ribs including their osteochondral junctions during 30 minutes. Sulphation rate was rather constant in growing female mice between 3 and 8 weeks age and then decreased markedly. Growing male mice exhibited an increase of sulphation activity until an age of 5 weeks and then a gradual decrease, similar to that of the females. Growth rates of both sexes were positively correlated with sulphate uptake activities. The form of the disappearance curve of incorporated sulphate suggested that the standard cartilage sample includes regions with different sulphate-metabolizing activities.

Hypophysectomy resulted in a decrease of sulphation rate to a level about 30% of the control values, which was constant for several weeks. Starvation of the animals likewise depressed the sulphation rate and after 72 hours starvation the values became similar to those after hypophysectomy. Dwarf mice had much lower sulphation activities than their heterozygous controls. Neither bovine GH nor human GH elevated the low sulphation activity of hypophysectomized mice but porcine and ovine GH-preparations had a pronounced and dose-dependent effect. Normal growing male and female mice showed no response to large doses of ovine GH. Ear, xiphoid and tracheal cartilage from hypophysectomized animals behaved as costal cartilage with respect to the reduction in sulphation rate, but GH restored the sulphation values to normal only in costal cartilage. A single dose of 1 mg ovine GH given to hypophysectomized mice at different times in relation to the sulphation experiment increased sulphate incorporation only after a time lag of more than 8 but less than 17 hours after injection of GH, whose effect lasted at least 38 hours.
FUNCTIONAL MICROANGIOPATHY IN ALLOXAN DIABETIC RATS
Authors: Scher Steven M. , Lawrence A. M.
Published: December 2008

Increased capillary permeability can be demonstrated in rats with recently acquired diabetes mellitus induced with alloxan. Alloxan injected non-diabetic rats failed to demonstrate such functional microangiopathology. Diabetic animals were characterized by more rapid transudation of intravenously administered bromphenol blue into intradermal wheals, more rapid loss of radioactive rat serum proteins from the vascular compartment and by the development of gross peripheral oedema or anasarca after intravenous administration of high molecular weight dextran. By contrast, non-diabetic rats, lacking increased vascular permeability, died in acute pulmonary oedema following intravenous colloid overload. It is proposed that functional pathology of the microvasculature characterized by increased capillary permeability may be significantly responsible for the special anatomic changes seen in human diabetic angiopathy.
DISAPPEARANCE IN PLASMA AND TISSUE UPTAKE OF RADIOACTIVITY AFTER AN INTRAVENOUS INJECTION OF [6,7-3H] NORETHYNODREL IN WOMEN
Authors: Laumas K. R. , Murugesan K. , Hingorani V.
Published: December 2008

After an iv injection of [6,7-3H] norethynodrel to six women, the disappearance of radioactivity in plasma and the uptake in endometrium, myometrium and cervix at different times have been studied. The plasma disappearance curve due to norethynodrel and its metabolites showed an initial rapid disappearance with an average half-life of 76 min and a subsequent slower disappearance with a mean half-life of 45 h. The average volumes of distribution V1 and V̄ are about 17 and 119 litres respectively. The disappearance curve has been analysed on the basis of a biexponential curve representing a model consisting of two compartments. The second average half-life of 45 h is much longer than that of other hormones studied and it also explains the effectiveness of norethynodrel for 24 h or longer. Disappearance of radioactivity in the erythrocytes followed a similar pattern of disappearance of radioactivity in plasma, however, the levels were lower.

The molar concentration of radioactivity was of the order of 14.5 × 10−9 m in the endometrium, 7.1 × 10−9 m in the myometrium and with the progress of time a higher uptake in the endometrium compared with the myometrium was found. Sub-cellular distribution showed 60–70% in the 105 000 g supernatant and 30% in the nuclear fraction of the endometrium. The significance of these results on the transport and action of norethynodrel is discussed.
EQUILIBRIUM CONCENTRATION OF [6,7-3H]OESTRONE AND [6,7-3H] 17β-OESTRADIOL METABOLITES IN FEMALE RAT TISSUES, DURING LONG-TERM INTRAVENOUS INFUSIONS
Authors: Hertogh R. De , Ekka E. , Vanderheyden I.
Published: December 2008

Equilibrium concentrations of oestrone (OE1) and 17β-oestradiol (OE2) metabolites in rat tissues were investigated by means of long-term infusions with varying amounts of labelled hormones.

3 hour-infusions were found sufficient to allow an equilibrium to be established between concentrations of metabolites in the plasma, liver, uterus, adrenals, fat and striated muscle and the infusion rate. In all these tissues, except the uterus, the equilibrium concentration was directly proportional to the rate of infusion. (The »concentration index«, i. e. the concentration per unit rate of infusion was constant). Hence, the results could be directly interpreted on a physiological basis. OE1 and OE2 were the major metabolites in all the tissues investigated, accounting for 47 to 78% of the total radioactivity in the plasma, 82 to 102% in the uterus, 40 to 48% in the liver, 80 to 120% in the adrenals, 53 to 85% in the adipose tissue and 66 to 93% in the muscle. Other fractions consisted of conjugates, mostly in the liver: 11 to 23% and in the plasma: 15 to%, of polar metabolites, mostly in the liver: 19% to 33%, or of non-polar metabolites, mostly in the adipose tissue: 5 to 17% and in the adrenals: 2 to 12%. The infused hormone was always predominant in all the tissues investigated except in the liver where the OE1/OE2 ratio was about 10, in either type of infusions. Hence, extensive interconversion between OE1 and OE2 was found to occur in that tissue. OE1 was taken up to a higher level than OE2 in the adipose tissue, whereas the reverse situation occurred in the uterus. In the latter tissue, the uptake was higher towards the end as compared to the early period of the cycle.

A »molar concentration index« (i. e. molar concentration per unit rate of infusion) of the infused hormone (either OE1 or OE2) was calculated. It ranged from 0.7 to 7 × 10−11 m/kg/ng/h, depending on the tissue investigated. Hence the OE2 concentration in these tissues under physiological conditions is not likely to exceed 10−9 m/kg.

The results are discussed with regard to the mechanism of hormone uptake in the tissues and the metabolism of the hormone, as assessed by urinary analysis.
ROLE OF NON-LUTEAL OVARIAN TISSUES IN THE MAINTENANCE OF PREGNANCY
Authors: Kraicer P. F. , Kisch E. S. , Nussbaum M. , Tandlich Ada
Published: December 2008

The maintenance of pregnancy by daily injections of progesterone from the time of nidation to term has been studied in rats following removal of the ovaries, removal of the active corpora lutea or pharmacological suppression of the corpora lutea with ergocornine. Progesterone was able to maintain normal embryogenesis following ergocornine or removal of corpora lutea but not in ovariectomized rats. This suggests that the non-luteal tissues of the ovary secrete hormones essential for the maintenance of gestation. Other observations include a high pre-implantation loss of embryos and an increase in the average conceptus weight not correlated with the litter size following removal of corpora lutea or of the ovaries. It is concluded that pregnancy is maintained by the combined activities of luteal and non-luteal ovarian tissues.
EXCRETION OF OXYTOCIN AND VASOPRESSIN IN HUMAN URINE
Authors: Frandsen Per , Jensen Svend Erik
Published: December 2008

A method has been developed for the estimation of oxytocin in human urine. The assay of oxytocin was based on the milk-ejecting activity measured in lactating rats following a preceding chromatographic concentration of oxytocin. Recovery experiments of oxytocin were performed, and the yield was about 70%. No endogenous milk-ejecting activity was detected, i. e. the excretion of endogenous oxytocin, if any, was less than 2 mU per 24 h. The 4 hour urine excretion of vasopressin and oxytocin was respectively about 5% and 0.2% of the subcutaneously injected dose.
RADIOIMMUNOASSAY OF URINARY HUMAN CHORIONIC GONADOTROPHIN
Authors: Arends J.
Published: December 2008

A description of a radioimmunoassay procedure for human chorionic gonadotrophin (HCG) in urine is presented.

The labelled hormone used is purified by electrophoresis on polyacrylamide columns, and several parameters are described for assessing the immunological reactivity of the final product.

Separation of free from antibody-bound hormone is accomplished by salting-out with ammonium sulphate, and a detailed account of the technique is given.

Interference with both the precipitation process and with the antigen – antibody reaction by non-hormonal constituents in urine can for all practical purposes be disregarded.

Application of the method for routine work on pregnancy urines shows good correlation with a bioassay method. Comparison of biopotency and immunopotency of HCG in urines from patients with trophoblastic diseases indicates a deviation from HCG in normal uncomplicated pregnancies, although this is not of sufficient magnitude to allow a differential diagnosis.

The inter-assay variation coefficient is about 18% and the sensitivity is good, but is limited in practice due to cross reactivity with luteinizing hormone.
COMPARISON BETWEEN β1–24 CORTICOTROPHIN DW-75 (SANDOZ) AND NATURAL CORTICOTROPHIN IN NORMAL ADULTS PRETREATED WITH DEXAMETHASONE
Authors: Helweg-Larsen Karin , Ulrich Jørgen
Published: December 2008

Three different ACTH preparations, a synthetic pentacosapeptide (DW-75, Sandoz), a synthetic tetracosapeptide (Synacthen®) and natural ACTH (Acton®) were administered to 11 normal young experimental subjects by intravenous injection. The biological activity was measured by the rise in plasma 11-OH-corticosteroids in the experimental subjects in whom the endogenous ACTH production was suppressed by means of dexamethasone.

The study of the stimulating effect on the adrenal cortex showed that both with regard to magnitude of the maximal response and the duration of the response, the three preparations had more or less the same activity per international unit as measured by Sayers' subcutaneous test. In addition, it was found with all three ACTH preparations that the intravenous injection of a dose between 1 IU and 2 IU gave a maximal rise in plasma 11-OH-corticosteroids.