bluuebunny/crossref_metadata_2025 huggingface dataset:
APPLICATION OF WICK CHROMATOGRAPHY TO THE RADIOIMMUNOASSAY OF PEPTIDE HORMONES AND OTHER POLYPEPTIDES
Authors: Bansal D. D. , Bajaj S. , Vallance-Owen J.
Published: December 2008

Wick chromatography has been used for assessment of iodination damage as well as for the separation of free from the antibody bound fraction of five polypeptides (insulin, A and B chains of insulin, glucagon and gastrin). The study shows that this technique should be destricted to basic polypeptides (insulin, B-chain of insulin and glucagon) and that the method cannot be applied to the radioimmunoassay of acidic polypeptides (gastrin and A-chain of insulin).
THE EFFECTS OF OESTROGENS ON THE LYSOSOMES OF RAT VAGINAL EPITHELIUM; A HISTOCHEMICAL STUDY
Authors: Nozawa Shiro , Kurihara Soju , Tsutsui Fumio , Watanabe Keiichi
Published: December 2008

Enzyme histochemical changes of the lysosomal enzymes (N-acetyl-β-glucosaminidase, β-glucuronidase and acid phosphatase) were investigated in the rat vaginal epithelium after oestrogen administration (oestradiol-17β and oestriol). The enhancement in the activity of the lysosomal enzymes and their diffuse appearance in the stratum granulosum of the vaginal epithelium was observed up to the cornification layer. Subsequently, the cornified layer and the uppermost epithelial cell layer were detached from the remainder of the epithelium. These phenomena were moderately well suppressed by the administration of cortisol, a lysosome stabilizer. The mechanism of formation of the cornified layer and its subsequent detachment is discussed with special reference to the lysosome labilizing effect of oestrogens.
DIENCEPHALIC INVOLVEMENT IN GENERALIZED LIPODYSTROPHY: RATIONALE AND TREATMENT WITH THE NEUROLEPTIC AGENT, PIMOZIDE
Authors: Corbin Alan , Upton G. Virginia , Mabry C. C. , Hollingsworth Dorothy R.
Published: December 2008

Our results indicate that generalized lipodystrophy (GLD) possesses as one of its manifestations, diencephalic neuroendocrinopathy. It has been postulated that deranged hypothalamic catecholaminergic mechanisms lead to the hypersecretion of hypophysiotrophic hormones that are detectable in the peripheral circulation. Initial treatment of a pre-puberal GLD patient with chlorpromazine was unrewarding. Chronic treatment of the same patients with the selective dopaminergic blocking agent, pimozide, eliminated corticotrophin releasing factor and luteinizing hormone releasing factor activities from the general circulation. Pimozide treatment ameliorated most of the distorted blood chemistry profile, led to return of facial subcutaneous fat, and elimination of cutaneous hyperpigmentation, fever, abdominal pain, hypertrichosis, oilness of hair, scaliness of skin and to a state of general well-being.

The results of these studies, structured within the framework of our present knowledge concerning hypothalamic neurotransmitter/releasing hormone physiology, support the concept of diencephalic involvement in GLD and may provide a guideline to effective therapy.
L-DOPA INDUCED GROWTH HORMONE SECRETION. COMPARISON WITH INSULIN TOLERANCE TEST, ARGININE INFUSION AND SLEEP INDUCED GH-SECRETION
Authors: Lucke C. , Höffken B. , Morgner K. D.
Published: December 2008

Plasma growth hormone (GH) levels following oral stimulation with 500 mg of L-dopa were determined and compared with those following arginine infusion, insulin tolerance test (ITT) and sleep induced secretion. Peak values following L-dopa were 13.3 ± 2.5 ng/ml and not different from those following arginine (16.4 ± 3. 3ng/ml) and those during nocturnal sleep (14.7 ± 1.2 ng/ml), but significantly less than GH peaks during ITT (24.9 ± 3.4 ng/ml). The rate of false negative results to L-dopa was 15% and seems to be higher than following other stimuli. – In contrast to the stimulatory effect in healthy subjects L-dopa had a suppressive effect on the GH levels in 4 patients with acromegaly.
INHIBITION BY PROPYLTHIOURACIL OF THE EXTRATHYROIDAL FORMATION OF TRIIODOTHYRONINE FROM THYROXINE
Authors: Bernal J. , del Rey F. Escobar
Published: December 2008

Thyroidectomized rats were injected daily with 1.9 μg/100 g body weight of 125I-labelled L-thyroxine (T*4). When isotopic equilibrium was attained, half the experimental group was injected twice with 2 mg of propylthiouracil at a 24 h interval. Animals were killed 7 h after the last dose and the amounts of T*4 and labelled iodide and T3 in peripheral tissues were determined after extraction and paper chromatography. The amount of T3 derived from T4 is significantly lower in tissues of rats receiving PTU.
RENAL TUBULAR LOCALIZATION OF PARATHYROID HORMONE INDUCED URINARY CYCLIC ADENOSINE 3′,5′-MONOPHOSPHATE
Authors: Scurry Murphy T. , Pauk George L.
Published: December 2008

Stop-flow studies were done in three dogs before and after a parathyroid hormone (PTH) infusion. The urine cyclic adenosine 3′,5′-monophosphate (cyclic AMP) to inulin U/P ratios did not change during the control stop-flow but with PTH rose sharply and significantly in the proximal tubular samples. The rise in cyclic AMP to inulin U/P ratios was correlated with the fall in phosphorus concentration in the same samples. The PTH induced urinary cyclic AMP enters the tubular fluid in the same proximal area of the tubule in which PTH is known to affect phosphate reabsorption.
HORMONE AND ENZYME ASSAYS IN PREGNANCY
Authors: Christensen Arne , Frøyshov Dag , Fylling Petter
Published: December 2008

A comparative study of human chorionic somatomammotrophin (HCS), placental cystine-aminopeptidase (P-CAP), progesterone and total urinary oestrogens (Oe) in pregnancies complicated with essential hypertension, mild or severe pre-eclampsia was performed.

No significant reduction in the 4 parameters could be demonstrated in cases complicated with essential hypertension.

In cases complicated with mild pre-eclampsia, however, a significant reduction was found in the HCS (P < 0.001) and P-CAP (0.01 <P < 0.02) but not in the urinary oestrogen values (0.1 < P < 0.2). A tendency to increased progesterone values could be demonstrated. Furthermore, in those cases of mild pre-eclampsia associated with a birth weight below the 10 percentile, the HCS and P-CAP assays proved to be more sensitive than the urinary oestrogen assay. No influence upon the progesterone levels was observed in these cases.

On the other hand, in pregnancies complicated with severe pre-eclampsia the HCS, P-CAP and urinary oestrogen values were significantly reduced (P < 0.001) but not the progesterone values (P > 0.5). Again, when severe pre-eclampsia was associated with low birth weight, HCS and P-CAP were the most reliable tests.

Based upon the HCS, P-CAP and the urinary oestrogen readings placental and foeto-placental scores were introduced.

The placental score provided valuable information concerning the viability of the infants but no additional information was obtained by the foetoplacental score. The results indicated that simultaneous measurements of HCS and P-CAP as a placental function test and total urinary oestrogens as a foeto-placental test contribute reliable information in pregnancies complicated with pre-eclampsia. The measurement of progesterone could not be recommended as a test for the placental function.
EFFECT OF PREGNANCY ON CYTOPLASMIC AND MITOCHONDRIAL ENZYMES IN HUMAN AND ANIMAL MYOMETRIUM
Authors: Geyer Helmut , Riebschläger Michael
Published: December 2008

An investigation was made on the influence of pregnancy on the specific activities of cytoplasmic (lactate dehydrogenase2), cytoplasmic malate dehydrogenase) and mitochondrial enzymes (glutamate dehydrogenase, mitochondrial malate dehydrogenase, cytochrome-c-oxidase) in the human and animal myometrium. The activities were related to DNA.

The specific activities of all the investigated enzymes increased. This rise in activity depended on the cellular localisation of the enzyme. The activity of all enzymes in one cellular compartment changed to the same extent. This change varied according to species. With regard to the human tissue, the increase of the cytoplasmic enzymes was larger than that of the mitochondrial enzymes. In the rat, however, a significantly larger increase of the mitochondrial enzymes was found.

The increase in the specific activities of the cytoplasmic enzymes in the human and rat was proportional to the protein-content and to the hypertrophy of the cells.

It was concluded that the number of mitochondria or their enzymatic activity increased in both species during pregnancy – in each species, however, to a different extent.

The pattern of the LDH-isoenzymes in the myometrium changed in the same manner in the human myometrium as in the rat. The percentage of M subunits of LDH compared to H subunits rose in both cases during pregnancy.
SECRETION OF DIHYDROTESTOSTERONE BY HUMAN PROSTATE IN BENIGN PROSTATIC HYPERTROPHY
Authors: Mahoudeau J. A. , Delassalle A. , Bricaire H.
Published: December 2008

Plasma levels of testosterone (T) and 5α-dihydrotestosterone (DHT) were determined by radioimmunoassay in 29 patients with benign prostatic hypertrophy (BPH) and in 56 control men of various ages. No significant difference was found in T, DHT nor DHT/T ratio between BPH and control subjects of similar age. Plasma DHT was higher in the prostatic than in the peripheral veins in 8/9 patients with BPH during laparotomy, indicating a prostatic secretion of DHT. No difference in the mean T nor the mean DHT was found in peripheral plasma before and after adenomectomy.
ADRENALECTOMY AND PITUITARY LH IN THE OOPHORECTOMIZED SUCKLED RAT
Authors: Landers E. , Wagner W. C.
Published: December 2008

Oophorectomy of suckled rats caused a significant increase in pituitary LH content (214 μg/mg vs 156 μg/mg fresh anterior pituitary). Oophorectomized non-suckled rats were even higher (267 μg/mg). Adrenalectomy of oophorectomized suckled rats resulted in a highly significant decrease in pituitary LH (71 μg/mg) while treatment with flumethasone (0.125 mg/day) resulted in pituitary LH levels of 177 μg/mg. Except for a significant decrease in body weight and adrenal gland weight in flumethasone treated rats, no other significant treatment effects were noted. These results confirm the interaction of adrenal and pituitary-gonadal function in the post-partum rat.
EFFECTS OF PITUITARY HORMONES AND CORTICOSTERONE ON LIPOLYSIS IN HYPOPHYSECTOMIZED RATS
Authors: Schillinger Ekkehard , Gerhards Erich
Published: December 2008

Removal of the pituitary gland in rats reduces basal lipolysis and greatly diminishes the lipolytic response of isolated fat cells towards L-epinephrine, ACTH and dibutyryl-cyclic-3′5′-AMP (DB-cAMP). Subcutaneous application for 5 days of ACTH, LH, FSH, GH and TSH, single or in combination, does not appreciably influence the responsiveness of fat cells towards lipolytic stimulation. Administration of a hypophyscal homogenate restores the sensitivity of fat cells of hypophysectomized rats. A similar effect is obtained with a combination of growth hormone and corticosterone. In hypophysectomized-adrenalectomized animals only the combination of growth hormone and corticosterone, but not a pituitary homogenate, produces a normal responsiveness of fat cells towards lipolytic stimuli.

It is concluded that both the pituitary and adrenal gland are intimately involved in the hormonal regulation of lipolysis in fat cells.
HYPERCALCAEMIA OF NEOPLASIA, PARATHYROID HORMONE AND VITAMIN D: STUDIES IN PARABIOSIS
Authors: Ponthier Roy L. , Rice Bernard F.
Published: December 2008

A localized transplantable Leydig cell tumour of the Fischer rat regularly produces lethal hypercalcaemia in thyroparathyroidectomized and intact hosts. The parabiosis model was used to explore and compare the hypercalcaemia producing action of this tumour, parathyroid hormone, and vitamin D. Administration of exogenous parathyroid hormone (10 and 20 units) to one thyroparathyroidectomized parabiont rat increased the serum calcium in both parabionts within 24 hours. Endogenous parathyroid hormone, studied by producing parabiosis between thyroparathyroidectomized rats and intact rats, raised the serum calcium of the thyroparathyroidectomized parabiont within 7 days in parabiosis, suggesting that endogenous parathyroid hormone is effective across the parabiotic union. Vitamin D3 (100 000 and 150 000 units) given to one parabiont of a thyroparathyroidectomized pair produced hypercalcaemia only in the injected parabionts. When the hypercalcaemia producing tumour was grown in thyroparathyroidectomized pairs for 3–6 weeks, severe hypercalcaemia and death occurred only in the tumour bearing parabiont. The serum calcium of the non-tumour parabiont did rise over a period of 3–6 weeks and was compatible with an approach to calcium equilibrium across the parabiotic union occurring over an extended period of time. These experiments suggest that the hypercalcaemia produced by the hormone secreted by the transplantable Leydig cell tumour resembles vitamin D3 rather than parathyroid hormone when studied in the parabiosis model. These results, when reviewed in light of the parabiosis model and other recent studies, suggest that this hypercalcaemia producing hormone is probably not parathyroid hormone or vitamin D or one of its metabolites, but is of small molecular weight and rapidly metabolized. Whether it represents a new hormonal substance not previously described or is a known substance whose properties have not been fully characterized, remains to be elucidated.
CHANGES IN THE CONCENTRATIONS OF OESTRONE AND OESTRADIOL IN THE PERIPHERAL PLASMA OF THE DOMESTIC HEN DURING THE OVULATORY CYCLE
Authors: Senior B. E.
Published: December 2008

The concentrations of oestrone and oestradiol were measured by radioimmunoassay in the peripheral plasma of hens during the ovulatory cycle and of non-laying hens. During the ovulatory cycle when blood samples were taken at 4 or 6 h intervals for 24 h, the peak concentrations of oestrone and oestradiol occurred within the 6 h period immediately preceding ovulation. After the final ovulation of the clutch the oestradiol concentration fell gradually and did not rise again near the time when the last egg of the clutch was laid. There was no significant change in the concentration of oestradiol over a 24 h period in non-laying hens. When blood samples were taken at 2 h intervals during the 12–14 h period before the expected time of ovulation the oestradiol concentration rose 8 h before ovulation and reached a peak level 6–4 h beforehand. It is suggested that oestradiol may be involved in the mechanism stimulating the release of luteinizing hormone required for ovulation.
THE GONADOTROPHIN RESPONSE TO SYNTHETIC GONADOTROPHIN-RELEASING HORMONE IN MALES IN RELATION TO AGE, DOSE, AND BASAL SERUM LEVELS OF TESTOSTERONE, OESTRADIOL-17β AND GONADOTROPHINS
Authors: Haug E. , Aakvaag A. , Sand T. , Torjesen P. A.
Published: December 2008

Three healthy male volunteers, 28–35 years old, were given an intravenous dose of 12.5, 25, 100, 200, 400, 800, and 1200μg of synthetic gonadotrophin-releasing hormone (LH/FSH-RH) in order to determine the dose-response relationship. The resulting log dose-response curve between interstitial-cell stimulating hormone (ICSH) and LH/FSH-RH was approximately linear between the 12.5 and 400 μg doses. The smallest dose that significantly (P < 0.05) increased the mean serum ICSH levels was 25 μg, and 400 μg of LH/FSH-RH was the minimum dose that produced the maximum ICSH response. The increments in serum follicle-stimulating hormone (FSH) were much less than the corresponding serum ICSH increments, and the log dose-response curve was not linear.

The effect of increasing age on the gonadotrophin response to LH/FSH-RH was studied in 50 male subjects, 20–89 years old, without evidence of endocrine disease. They were given 100μg of LH/FSH-RH each. A reduction in the mean response for both ICSH and FSH was observed after the age of 70, and this was most clearly seen in the group of males 80–89 years old. Therefore the interpretation of the LH/FSH-RH tests, at least in males, will require age-specific ranges of normal ICSH and FSH responses.

Serum testosterone levels were determined in 48 of the male subjects investigated, and the mean serum levels remained within the same range (above 0.60 μg/100 ml) from the age of 20 to the age of 79 years. In the age group 80–89 years, the mean serum testosterone was reduced to 0.39 μg/100 ml. Serum oestradiol-17β was determined in 44 of the males, and there were no significant differences between the mean levels of the different age groups (45–55 pg/ml).

There was no significant age-dependent change in basal serum ICSH and FSH levels.

It is concluded that there is a parallel decrease in serum testosterone levels and in serum gonadotrophin responses to LH/FSH-RH after the age of 70. These findings would suggest that the decrease in serum testosterone levels is the result of reduced stimulation of the testicular tissue with endogenous gonadotrophins.
DISCREPANCIES IN PLASMA LH ACTIVITIES AS MEASURED BY RADIOIMMUNOASSAY AND AN IN VITRO BIOASSAY
Authors: Qazi M. H. , Romaní P. , Diczfalusy E.
Published: December 2008

Using a highly sensitive in vitro bioassay system, luteinizing hormone activity has been measured in parallel with radioimmunoassays in postmenopausal plasma and plasma obtained from women in various phases of the menstrual cycle (follicular, mid-cycle, luteal). Biological and immunological activities were measured directly in plasma samples without any chemical manipulation. The biological activity (B) was always higher than the immunological activity (I); the B/I ratio varied from 2.1 to 14.0. Gel filtration of pooled plasma samples through Sephadex G-100 revealed major discrepancies in each physiological state when immunological and biological activities were measured in each fraction. The biological activity was eluted as a single peak behind the elution volume of bovine serum albumin, but in front of the elution volume of chymotrypsinogen. It was invariably preceded by a small hump. The immunological activity was spread all over the chromatogram. Areas of immunological activity without any biological activity were located on either side of the biologically active fractions, both in the high molecular weight range (including the void volume) and in the low molecular weight range. The biological LH activity recovered following fractionation on Sephadex G-100 was in close agreement with that loaded, whereas the immunological activity recovered following gel filtration exceeded the loaded activity by a factor of 6–8. In the various physiological states, 11 to 44 % of the total immunological activity recovered was not associated with any biological activity. Furthermore, there was a marked variation in the ratio of biological to immunological activities of those fractions which contained biological activity.

It is suggested that the specificity of current RIA methods could be improved significantly by preparing antisera which react only with biologically active LH species.
CYCLIC AMP IN ISOLATED CORPORA LUTEA OF THE RAT: INFLUENCE OF GONADOTROPHINS AND PROSTAGLANDINS
Authors: Herlitz Hans , Hamberger Lars , Rosberg Sten , Ahrén Kurt
Published: December 2008

Corpora lutea were isolated from 34–39 day-old rats injected with 10 IU PMSG when 30 days old leading to ovulation early in the morning of day 33.

Three day-old corpora lutea were incubated in Krebs bicarbonate buffer containing 5.5 mmol/l glucose. Addition of LH to the incubation medium increased accumulation of cyclic 3′, 5′-AMP (cAMP) in a dose-dependent way in the isolated tissue and caused a release of cAMP to the incubation medium. cAMP was determined by the protein binding technique of Gilman (1970). There was a continuous rise in the levels of cAMP in both tissue and medium with increasing incubation time in presence of LH. Neither prolactin nor FSH could influence the cAMP levels in the corpus luteum tissue in vitro. Prostaglandin E2 (PGE2) caused a stimulation with respect to cAMP formation. The time-course of the PGE2 effect was quite different from that of LH: PGE2 gave maximal tissue cAMP content within 5–15 min with decreasing levels thereafter, compared to the continuous rise with LH stimulation for at least 1 h.

In experiments performed on corpora lutea of different ages (1–7 day-old), tissue levels of cAMP were determined after incubations with LH or PGE2. There was a dramatic increase of tissue cAMP in the very young corpus luteum and a marked decrease in sensitivity to LH with increasing age of the corpus luteum with a very small but still significant effect on the old corpora lutea. The stimulatory effect of PGE2 on cAMP levels on the other hand showed only a minimal change.

A few experiments on the first generation of corpora lutea from mature cycling rats are also reported for comparison.
THE EFFECT OF SHORT AND LONG TERM HUMAN CHORIONIC GONADOTROPHIN (HCG) ADMINISTRATION ON PLASMA TESTOSTERONE LEVELS IN KLINEFELTER'S SYNDROME
Authors: Smals A. G. H. , Kloppenborg P. W. C. , Benraad Th. J.
Published: December 2008

The effect of acute (1500 IU/day for 3 days) and chronic HCG administration (1500 IU, 3 times weekly) on plasma testosterone levels in patients with Klinefelter's syndrome was compared with the response observed in patients with hypogonadotrophic eunuchoidism and in eugonadal male controls. Basal plasma testosterone levels in the Klinefelter patients were significantly lower than in the control subjects and significantly higher than in the patients with secondary hypogonadism. In all but one Klinefelter patient the plasma LH levels were markedly elevated even in the presence of normal testosterone levels. No significant correlation could be demonstrated between the plasma testosterone concentrations and the LH levels in the Klinefelter patients. Short term HCG administration resulted in a significant increase in the plasma testosterone levels in each of the 3 groups studied, independent of the basal value. The absolute increase in the Klinefelter patients was quantitatively comparable to that in the patients with secondary hypogonadism, but significantly lower than in the eugonadal controls. During long term HCG treatment the plasma testosterone levels definitely increased in both patient groups, but remarkably in the Klinefelter patients testosterone levels tended to decrease on continuing treatment, though in most patients testosterone levels remained higher than the pre-treatment values. The data on the effect of acute and chronic HCG administration on plasma testosterone levels in this study illustrate again that Leydig cells in Klinefelter's syndrome still retain a functional reserve, though less than in eugonadal males.
TESTOSTERONE METABOLISM IN CULTURED HYPERPLASIA OF THE HUMAN PROSTATE
Authors: McMahon M. J. , Butler A. V. J. , Thomas G. H.
Published: December 2008

Slices of human benign prostatic hyperplasia were maintained in organ culture in the presence of [3H]- or [14C]testosterone.

Explants concentrated radioactivity from the culture medium, although this effect was depressed by the inclusion of foetal calf serum in the medium.

Testosterone was metabolised to products with chromatographic mobilities corresponding to androstanediols, androsterone, dehydroepiandrosterone, and androstanedione. The principal metabolite was identified as 5α-dihydrotestosterone, and small amounts of testosterone and androstenedione were also found.

As culture time increased from 1 to 6 days there was a diminution in the proportion of 5α-, and an increase in 17-ketometabolites, indicating a swing to a less physiological pattern of testosterone metabolism by the aging culture. Testosterone metabolism was thought to provide a sensitive index by which to evaluate changes in culture conditions.
SERUM GROWTH HORMONE AND GONADOTROPHINS AND URINARY STEROIDS IN ADOLESCENT GIRLS
Authors: Kantero Riitta-Liisa , Wide Leif , Widholm Olof
Published: December 2008

In a series of 146 school girls and nurses in training, the serum levels of growth hormone (GH), follicle stimulating hormone (FSH), and luteinizing hormone (LH) were determined in the afternoon after 4–5 hours' fast. Blood specimens of the menstruating girls were taken on the 7th to 10th day from the beginning of the preceding menstrual bleeding. From the 24-hour urines collected on the previous day the excretion of total oestrogens, 17-ketosteroids and 17-hydrocorticosteroids was determined. The series was grouped according to skeletal age which varied from 8.0 years to the adult stage.

In the girls aged 11-13 years the GH curve showed a significant rise parallel with the mean height curve. The FSH values at the early age of 8–9 years corresponded to the follicular phase of adult women. The LH level increased steeply up to the 15th year of bone age and the mean values of adults were found to be about three times those of childhood. The excretion of total oestrogens and 17-ketosteroids increased steadily up to the full maturity of skeletal age. In the oldest group of girls the excretion was about four times that of the group aged 8–9 years. The hormonal maturation seems to continue until late puberty. The 17-OHCS, on the other hand, reached the adult level in the group with a bone age of 13 years. The excretion of 17-OHCS shows a steep rise from the age of 11 to 12 years and this acceleration of excretion perhaps exceeds the increase in surface area.
VIRILIZING ADRENOCORTICAL ADENOMA RESPONSIVE TO GONADOTROPHIN
Authors: Blichert-Toft M. , Vejlsted H. , Kehlet H. , Albrechtsen R.
Published: December 2008

A virilizing adrenocortical adenoma was demonstrated in a young female. Urinary 17-ketosteroid excretion and subfractions, plasma testosterone level and urinary 17-ketogenic steroid excretion were markedly increased. Dehydroepiandrosterone was the main constituent of the androgen excess. Otherwise the adrenocortical function was found to be normal as evaluated from measurements of cortisol, corticosterone and their metabolites under basal conditions and during dynamic tests.

The androgen excess showed an unexpected response to trophic hormones. Human chorionic gonadotrophin stimulation resulted in a pronounced increase in androgen production, whereas no gonadotrophin-dependency could be demonstrated by means of the oestrogen suppression test. Similarly, no corticotrophin-dependency could be demonstrated by corticotrophin stimulation and suppression tests.

Removal of the tumour resulted in normalization of the androgen production and no abnormal response upon human chorionic gonadotrophin administration could now be found.

The inappropriate response of tumours to trophic hormones is discussed. It is concluded that the reliability of stimulation and suppression tests in determining the site of excessive androgen production should be accepted with reservation.
URINARY EXCRETION OF FREE CORTISOL IN IMPAIRED RENAL FUNCTION
Authors: Sederberg-Olsen P. , Binder C. , Kehlet H.
Published: December 2008

Total cortisol and free, non protein-bound cortisol in plasma and urinary excretion of unconjugated free cortisol were measured during iv infusion of cortisol at varying dose rates in eight patients with impaired renal function. The results showed that free urinary cortisol decreased with decreased glomerular filtration rate (GFR), also compared to free cortisol level in plasma. An increase in free cortisol in plasma had no influence on GFR. It is concluded that determination of free urinary cortisol, otherwise useful in diagnosing Cushing's syndrome, may be of less value in patients with impaired renal function.
THE IN VIVO METABOLISM OF CORTISOL AND CORTICOSTERONE BY THE MACAQUE MONKEY (Macaca fascicularis)
Authors: Setchell K. D. R. , Shackleton C. H. L.
Published: December 2008

[4-14C] Cortisol was administered intramuscularly to one adult female macaque monkey, MF3 (Macaca fascicularis). To adult female macaque monkey, MF4, [4-14C]corticosterone was administered intramuscularly. Urine samples were collected and the metabolites excreted identified using gas chromatography, radio-gas chromatography and gas chromatography-mass spectrometry.

The principal metabolites of cortisol were identified as glucuronide conjugates of 11-oxygenated-17-oxosteroids. The excretion of tetrahydrocortisol and tetrahydrocortisone relative to the other corticosteroid metabolites was low compared with that of man. Two compounds, 3β-cortol and 3β-cortolone not normally present in human urine were identified in the urine from this species.

The principal metabolites of corticosterone were glucuronide conjugates of hexahydroCompound A and hexahydrocorticosterone. Two unidentified radioactive compounds were also present.
STUDIES ON ISOLATED RAT ADRENAL CELLS I. CONTINUOUS FLOW AND BATCH INCUBATIONS
Authors: Falke H. E. , Degenhart H. J. , Abeln G. J. A. , Visser H. K. A. , Croughs R. J. M.
Published: December 2008

A procedure for the continuous flow incubation of isolated adrenal cells is described. In this way the advantages of continuous flow incubations of adrenal tissue are combined with those of isolated adrenal cells. Suspensions of isolated adrenal cells were prepared by a modification of the collagenase method. A sigmoid dose-response curve was obtained when these cells were incubated with ACTH in batch incubations. Under these conditions (in the presence of 1 mU ACTH/ml) the corticosterone production rate remained constant during at least 240 min. This production rate was linearly related to the number of cells. Pre-incubation of the cells during 3 h resulted in an increased response to ACTH. In continuous flow incubations without ACTH the corticosterone production was negligible. With 100 μU ACTH/ml corticosterone production increased sharply after a short lag period. A maximum was reached after 60–75 min followed by a slow decrease. Cells pre-incubated in the continuous flow apparatus had a slightly diminished ACTH response without loss of affinity to ACTH. The continuous flow incubation of isolated adrenal cells offers new possibilities for the dynamic study of steroid biosynthesis in vitro. The method may also be valuable to study processes in a wide variety of other tissues.
HYPOTHALAMIC-PITUITARY FUNCTION IN ANOREXIA NERVOSA
Authors: Frankel R. J. , Jenkins J. S.
Published: December 2008

Hypothalamic-pituitary function was studied in 4 patients with anorexia nervosa of different degree of severity before and after refeeding. Gonadotrophin secretion was low in all subjects and improved in two after a prolonged period of feeding. In one subject there was a failure of pituitary gonadotrophin secretion after the administration of hypothalamic gonadotrophin-releasing hormone but the response was restored to normal after treatment. Thyroid function was reduced in one patient only but returned to normal after intravenous therapy for 6 days. Growth hormone and cortisol levels were elevated in all patients, and in one severe case the growth hormone values were extremely high. There was also a disturbance of the hypothalamic control of growth hormone and pituitary-adrenal function, which returned to normal after refeeding. One patient with severe hypothermia was resistant to the administration of a pyrogen, but developed a normal febrile response after treatment.
THE ANTIGENIC DETERMINANT OF THE LH-RELEASING HORMONE FOR THREE DIFFERENT ANTISERA
Authors: Arimura A. , Sato H. , Coy D. H. , Worobec R. B. , Schally A. V. , Yanaihara N. , Hashimoto T. , Yanaihara C. , Sukura N.
Published: December 2008

Antigenic determinants of LH-releasing hormone (LH-RH) were investigated by testing the cross-reaction of LH-RH analogues and fragments in LH-RH radioimmunoassay (RIA) systems using 3 different antisera against the LH-RH decapeptide. Rabbit antiserum No. 419 was generated against LH-RH adsorbed on polyvinylpyrrolidone (PVP). Antisera Nos. 710 and 742 were produced by immunizing rabbits with LH-RH conjugated either with bovine serum albumin through its C-terminus, or with human serum albumin through the N-terminus, respectively. For antiserum No. 419, the N-terminal (pyro)-glutamic acid and/or histidine in positions 1 and 2 of LH-RH, respectively, were found to enhance the antigen-antibody interaction, but were not indispensable for it. Similarly, the C-terminal amide and glycine-NH2 did not play a major role in these interactions. The LH-RH heptapeptide fragment, corresponding to amino acid sequence from positions 3 to 9, showed a cross-reactivity in this RIA system with LH-RH, although greater amounts than those of cold LH-RH were required for a comparable inhibition of binding of labelled LH-RH. For antiserum No. 710, the LH-RH hexapeptide fragment corresponding to positions 2 to 7 showed considerable cross-reac

tivity. Histidine in position 2 played an important role but neither the amide group nor the glycine amide group at the C-terminus were essential. For antiserum No. 742, the C-terminal tetrapeptide-amide fragment of LH-RH showed considerable cross-reactivity in the LH-RH, the amide moiety itself being of crucial importance. These antisera may be useful in investigating peptides related to LH-RH in biological materials.
CORTICOSTEROIDS IN NOCTURNAL BLOOD PLASMA OF COWS IN THE FIELD RELATED TO STAGE OF LACTATION AND PLASMA ACETOACETATE
Authors: Blom Anne Kristine , Halse K.
Published: December 2008

Two materials of nocturnal blood plasma samples, each representing more than 100 cows unselected with respect to health condition, showed acetoacetate variations within a wide range, overlapping with the range for animals with clinical ketosis. Within each of the two materials a statistically significant (P < 0.001) negative correlation (r = −0.30) was obtained between plasma corticosteroids and log acetoacetate.

The lowest corticosteroid and the highest acetoacetate averages were obtained for animals which were within the first month of the lactation cycle. With increasing time post-partum the two components varied inversely, a maximum in corticosteroids coinciding with a minimum in acetoacetate in cows which had calved 2½ to 3 months before the date of sampling. At this stage the mean plasma sugar was about 10 per cent higher than in early lactation.

The corticosteroid ranges for different levels of acetoacetate overlapped extensively. Still cows with acetoacetate ≤ 0.6 mg/100 ml gave significantly higher corticosteroid averages than the rest of the material. But in animals exceeding the 0.6 mg limit no relationship was found between the severity of the ketonaemia and the steroid level. Thus, no difference was obtained between cows with clinical ketosis and animals which were moderately ketonaemic.

Some change in adrenal function may be associated with the development of ketonaemic conditions in cows. But no proof has been provided of a direct adrenal involvement in the transformation of a case of subclinical ketosis to the clinical stage.
ANDROGENIC PROPERTIES AND ADRENAL DEPRESSANT ACTIVITY OF MEGESTROL ACETATE OBSERVED IN CASTRATED MALE RATS
Authors: Tisell Lars-Eric , Salander Håkan
Published: December 2008

Megestrol acetate (17α-acetoxy-6-dehydro-6-methylprogesterone), a synthetic steroid with high progestational activity, is used in oral contraceptives but also in the treatment of prostatic diseases in man. To investigate whether megestrol acetate has any androgenic properties the growth of the ventral and dorsolateral prostate, the coagulating glands and the seminal vesicles was studied morphologically in castrated rats treated with megestrol acetate and in non-treated castrated rats. The effect of megestrol acetate on the body weight, the levator ani muscle and the adrenals was also studied. Megestrol acetate was administered in daily doses of 0.02 mg, 0.2 mg, 2.0 mg or 20.0 mg for a period of 21 days. Megestrol acetate in the two higher doses retarded growth and gave a low weight for the levator ani muscle at autopsy indicating an anti-anabolic or catabolic action of megestrol acetate in high doses. Megestrol acetate in daily doses of 0.2, 2.0 and 20.0 mg caused an involution of the adrenal glands. After the two higher doses the weight of the adrenals amounted to only about a third of that of the untreated rats.

Megestrol acetate in the lower doses had no demonstrable effect on the growth of the accessory reproductive glands. After the two higher doses of megestrol acetate some growth of the dorsal part of the dorsolateral prostate and of the coagulating glands was observed. Only the seminal vesicles exhibited complete morphological criteria of an androgenic stimulation and then only after the largest dose of megestrol acetate. The investigation shows that megestrol acetate has weak androgenic properties which are apparent at a dose per kg body weight approximately 200 times greater than that used in the treatment of prostatic diseases in man.
OVARIAN FUNCTION DURING TREATMENT WITH LYNOESTRENOL 0.5 mg DAILY
Authors: Wiese Jørgen
Published: December 2008

Twenty-four women, all with normal ovulatory menstrual periods, were studied during in total 64 cycles on lynoestrenol 0.5 mg daily and during 11 control cycles. The 24-hour urines were collected for assay of total oestrogens on the 21st, 22nd and 23rd days of viz. the 1st, 4th, 6th and 12th cycles and for pregnanediol on the 22nd and 23rd days of the same cycles. Oestrogen excretion was at the same level during treatment and control cycles. All the control cycles and 9 and 14% respectively of the treatment cycles were ovulatory judging by the plasma progesterone and the pregnanediol excretion. On the basis of the latter parameter there was a certain luteal activity during 36 % of the treatment cycles. In all cycles but one, however, the plasma progesterone and the pregnanediol excretion were lower than during the control cycles. It must be assumed, therefore, that lynoestrenol in a dose of 0.5 mg daily inhibits the ovulation or function of the corpora lutea in practically all cases.
OVULATION INHIBITION WITH HUMAN CHORIONIC GONADOTROPHIN
Authors: Friedrich F. , Kemeter P. , Salzer H. , Breitenecker G.
Published: December 2008

Eight women with regular menstrual cycles were treated daily during 9 cycles with HCG (Human Chorionic Gonadotrophin) 3000 or 5000 IU daily for a period of 4–7 days. This treatment was started between the 1st and the 6th day after the onset of menstruation. Control of the treatment cycles was performed by basal body temperature, pregnanediol serial estimations, endometrial biopsies and in addition in 5 treatment cycles by radio-immunological assay of oestradiol-17β (Oe2), progesterone, LH and FSH from the serum at intervals of 1 to 3 days. In 6 of these cycles where treatment started on the 4th day or later, ovulation was inhibited (2 cycles) or postponed (4 cycles) to the 24th–46th day. In these 6 treatment cycles the progesterone and pregnanediol increase during HCG treatment was poor or absent. The typical Oe2 increase of the normal menstrual cycle was impaired. In the 3 remaining cycles where treatment was started on the 1st, 2nd and 4th day, we observed during HCG treatment increases in Oe2 and progesterone serum values similar to that found during corpus luteum activity, and menstruation from a secretory endometrium between the 13th–19th day of the cycle. The histologically examined ovaries of one woman who was treated with HCG from the 2nd to the 6th day of the cycle showed distinct Iuteinization of the theca interna of all tertiary follicles and a beginning degeneration of the granulosa. These findings give support to the hypothesis that the luteinization of the theca interna leads to degeneration of the tertiary follicles thereby causing ovulation inhibition or postponement of ovulation.
EFFECT OF INDOMETHACIN, AN INHIBITOR OF PROSTAGLANDIN BIOSYNTHESIS ON THE LENGTH OF PSEUDOPREGNANCY IN RATS AND HAMSTERS
Authors: Lau I. F. , Saksena S. K. , Chang M. C.
Published: December 2008

Pseudopregnancy (PSP) in rats was significantly lengthened after subcutaneous administration of 0.2–1.0 mg indomethacin/day, starting on day 5 of PSP. Injections of 1 mg indomethacin on days 5 and 6 or days 7 and 8, as well as a single injection of 2 mg indomethacin on day 6, also produced a significant increase of the duration of PSP. In hamsters, doses of 300–500 μg indomethacin per day starting on day 5 of PSP did not alter the length of PSP, but a significant prolongation varying from 11 to more than 30 days was observed after a treatment with 1 mg of indomethacin daily. It was further observed that the ovaries of hamsters autopsied on day 31 of PSP were very small and the uteri resembled those of ovariectomized animals. It is concluded that indomethacin lengthens pseudopregnancy in both rats and hamsters and that the latter species showed a more variable response. It seems that chronic administration of indomethacin in hamsters caused a long-lasting suppression of ovarian hormone production.
CHANGES IN LABELLING OF RIBONUCLEIC ACID SPECIES FRACTIONATED FROM THE RAT VENTRAL PROSTATE IN RESPONSE TO TESTOSTERONE
Authors: Isotalo Anja , Santti R. S.
Published: December 2008

The total RNA isolated at various times up to 24 h after testosterone administration from the ventral prostate of castrated rats, was labelled either by injecting 3H-orotic acid directly into the ventral prostate 6 h before the animals were killed, or by incubating prostatic tissue in vitro with 3H-uridine for 20 to 60 min. The isolated RNA was separated into tRNA, ribosomal RNA (Q1 RNA) and two DNA-like RNA fractions (Q2) and TD RNA) by chromatography on methylated albumin kieselguhr (MAK) columns, and the fractions were further analysed by sucrose gradient centrifugation.

Testosterone given into castrated animals for 12 h, stimulated the labelling of all main fractions. The radioactivity of TD RNA after a 60-min incubation period in vitro with 3H-uridine was approximately twice that seen in the castrated rat, while there was a 3.1- and 2.2-fold increase in the radioactivity of the Q1 and Q2 RNA fractions respectively. Kinetics of incorporation of 3H-uridine into different RNA fractions revealed that the hormone facilitated the labelling of the TD RNA fraction relatively more than that of the Q2 fraction. The injection of 3H-orotic acid into the ventral prostate labelled the Q1 RNA preferentially. More than 60 % of the recovered radioactivity was found in Q1 RNA (as 18 and 28 S). Testosterone increased markedly (9.4-fold) the labelling of this fraction. It was concluded that testosterone has an activatory effect on the production of ribosomal RNA, and the bulk of the testosterone effect on the total RNA labelling is to be found in this fraction. Furthermore, it seems likely that testosterone also stimulates both the synthesis and processing of DNA-like RNA.

When actinomycin D was given 2 h before the hormone administration in a dose of 25 μg per 100 g of body weight, there was no noticeable increase in the labelling of any fraction above the level seen in the untreated castrated rat. There is evidence that testosterone exerts some effects on the labelling of proteins with radioactive amino acids and 14C-glucose metabolism in the absence of that fraction of the total RNA synthesis which is sensitive to a low dose (25 μg per 100 g of body weight) of actinomycin D (Isotalo & Santti 1972). In this way it may be concluded that the major changes of the RNA synthesis after testosterone administration are likely to be secondary to the protein synthesis and glucose metabolism, or the hormone exerts its anabolic effect on prostatic cells at different sites and by different modes of action, each of which can be operated independently.
THE RELATIVE ROLES OF IODINATION AND IODOTHYRONINE CONTENT ON THYROGLOBULIN STABILITY
Authors: Fukuda Hitoshi , Greer Monte A.
Published: December 2008

We have independently varied the degree of iodination and of iodothyronine formation over a wide range by acutely administering various doses of perchlorate and/or methimazole to severely iodine-deficient rats 30 min before giving 131I− with graded quantities (1–100 μg of 127I−). Thyroids were removed 4 h later and the soluble protein analyzed for labelled iodoamino acid composition and with sucrose density gradient ultracentrifugation. Since the total thyroid iodine content before administering 127I− was less than 1 μg, calculation of the degree of iodination and iodothyronine content of the labelled Tgb could be made from the known specificity of the injected labelled iodide. Newly organified iodine ranged from < 0.1 to 1.4 μg/thyroid and labelled iodothyronines from < 5 to 962 pmoles/thyroid. Both the degree of iodination and iodothyronine content varied directly with Tgb stability in the absence of inhibitors. But when Tgb iodination was kept constant, Tgb stability at pH 10.1 varied directly with iodothyronine content. When iodothyronine content was kept constant, Tgb stability was independent of the degree of iodination. Correlation of stability with iodothyronine content was highly significant (r = 0.79, P < 0.001) but not of stability with iodine content (r = 0.49, P > 0.05). We conclude that the primary determinant of Tgb stability in mild alkali is the iodothyronine content and not the degree of iodination of the protein. The increased Tgb stability may be induced by coupling between iodotyrosil residues of different 12 S subunits rather than between residues of the same 12 S subunit.
THE DIAGNOSTIC VALUE OF A RADIOIMMUNOASSAY FOR PARATHYROID HORMONE IN HUMAN SERUM
Authors: Almqvist Sven , Hjern Bo , Wästhed Birgitta
Published: December 2008

A radioimmunoassay for the measurement of immunoreactive parathyroid hormone (PTH) in human serum is described. The assay is based on the ability of human parathyroid hormone (h-PTH) to compete with 125I-labelled bovine parathyroid hormone (b-PTH) for binding to a guinea-pig antiserum directed against b-PTH. The linear part of the standard curve was parallel with dose response curves for anti-b-PTH serum reacting with dilutions of sera from patients with primary hyperparathyroidism and from h-PTH purified from human parathyroid adenomas, indicating that levels of immunoreactive PTH could be expressed as b-PTH equivalents. The range in 62 healthy blood donors was 1.1–2.5 ng b-PTH Eq./ml. The reproducibility was satisfactory, and the sensitivity permitted the measurement of PTH concentrations down to 0.8 ng b-PTH Eq./ml. No cross-reaction with h-CT, h-STH or h-ACTH was observed.

The clinical value of the assay has been considered in a number of patients with various disorders of calcium metabolism, diagnosed and treated conventionally. About 80 per cent of patients with primary hyperparathyroidism had elevated PTH levels on one or more occasions before surgery. In patients with chronic renal failure of other aetiology than primary hyperparathyroidism the levels were usually far higher. Patients with primary hyperparathyroidism and increased S-creatinine had higher PTH levels than those with normal S-creatinine. After parathyroidectomy all previously increased PTH levels became normal or low. High PTH concentrations were found in 3 patients with normocalcaemic hyperparathyroidism who at operation were shown to have parathyroid adenomas. However, in normocalcaemic patients there were also some falsely elevated PTH values which limit the diagnostic value of the assay in this group of patients. Low PTH values were observed in patients with hypercalcaemia due to malignant disorders, indicating that PTH determination may be of some value in the diagnosis of patients with hypercalcaemia of unknown origin.