Quantitative electron microscopic studies on exocytosis and endocytosis in the thyroid follicle cell. The effect of TSH (0.5 U), given iv to T4-treated rats, on the distribution of membranes in the apical part of the thyroid follicle cell was investigated by electron microscopic morphometry. In unstimulated cells, the membrane surface area of exocytotic vesicles (Ev) was slightly larger than that of the apical plasma membrane; the latter had an estimated surface area of 180-190 micron2/follicle cell. Three phases in the action of TSH could be discerned. A first phase (up to 5 min) was dominated by exocytosis; the membrane surface area of Ev decreased by about 40% and the apical plasma membrane showed a corresponding increase; pseudopods were rare. A second phase (5-20 min) was characterized by an increase in the membrane surface area of pseudopods while the area of Ev and apical plasma membrane decreased. A third phase (20-30 min) was characterized by a decrease in the membrane surface area of the pseudopods and an equivalent increase in the membrane surface area of colloid droplets. The total membrane surface area measured was about the same in all groups of rats. The membrane surface area of endocytotic structures (pseudopods and colloid droplets) at 20 and 30 min was about the same as that of the apical plasma membrane in unstimulated cells. The present results are consistent with the hypothesis that after stimulation of the thyroid follicle cell, membrane is first transferred from Ev to the apical plasma membrane. From the enlarged apical plasma membrane, pseudopods are formed. Membrane from pseudopods is then internalized in the form of colloid droplets.


Selective inhibition of the secretion of triiodothyronines from the perfused canine thyroid by propylthiouracil. In order to assess a possible influence on the relative proportions of T4, T3, and rT3 in thyroid effluent, the thyroid-blocking agents, propylthiouracil (PTU) and methylmercaptoimidazol (MMI), were administered to perfused canine thyroids. The dog has two distinct thyroid lobes which were both perfused independently, one acting as a control for the other. T4, T3, and rT3 in thyroid effluents and thyroid hydrolysates were measured radioimmunologically. During control perfusion the T4:T3 and T4:rT3 ratios in thyroid effluent were considerably lower than in thyroid hydrolysates [t4:t3 (wt/wt), 6.1 +/- 2.0 vs. 12.8 +/- 4.4; T4:rT3, 23.0 +/- 5.9 vs. 69.7 +/- 29.1; mean +/- SD; n = 8; P less than 0.001], i.e. T3 and rT3 were secreted preferentially to T4. Infusion of PTU (1 mmol/liter) induced a gradual increase in both the T4:T3 and T4:rT3 ratios in thyroid effluent. The variation in the T4:rT3 ratio appeared somewhat earlier than that in the T4:T3 ratio. Both ratios eventually approached the ratios in thyroid hydrolysates. The change in ratios was caused by a PTU-induced decrease in the release of T3 and rT3 while the release of T4 was practically unaffected. In contradistinction, infusion of MMI did not affect the T4:T3 or T4:rT3 ratios in thyroid effluent. As PTU but not MMI inhibits T4 deiodination in peripheral tissues, the results of the present study suggest that part of the T3 and rT3 secreted from the thyroid stems from T4 deiodinated during the secretion by a mechanism similar to the T4-deiodinating processes in peripheral tissues.


Suckling stimulus, lactation, and suppression of ovulation in the rat. Lactation pseudopregnancy in rats suckling a 5-pup litter lasted 22.0 +/- 0.4 days (mean +/- SEM; n = 11). By day 13 of lactation (day 1 of lactation = day of parturition), the continuation of lactation pseudopregnancy was dependent on the suckling stimulus, as litter removal on day 13 resulted consistently in ovulation on day 16. Measurements of various hormones before and after litter removal revealed high concentrations of progesterone and PRL during lactation and a rapid drop of both hormone concentrations after litter removal. Lactation pseudopregnancy in rats suckling a 10-pup litter lasted 26.1 +/- 0.9 days (n = 16). After litter removal on day 13 of lactation, the lactation pseudopregnancy continued for a further 7- to 11-day period, as evidenced by daily vaginal smears which remained mucified during that period. Measurements of hormone concentrations revealed continuously high concentrations of PRL before litter removal and a pattern of PRL secretion characterized by at least two diurnal peaks during the first days after litter removal. Progesterone concentrations decreased by 50% after litter removal, but the levels then remained constant and well above those found after the removal of 5-pup litters. It is argued that the different response to litter removal on day 13 of lactation between rats suckling 5 or 10 pups is due to the initiation of PRL peaks in rats with 10-pup litters: these PRL peaks are able to maintain luteal function for some period. It is further argued that the initiation of PRL peaks in rats with 10-pup litters is due to the high blood concentrations of progesterone at the time of litter removal compared to those of rats with a 5-pup litter.


Human parathyroid hormone: synthesis and chemical, biological, and immunological evaluation of the carboxyl-terminal region. The carboxyl-terminal region of human parathyroid hormone (hPTH) was synthesized by the solid phase method. The 32-amino acid fragment, hPTH-(53-84), was prepared for two reasons: 1) to produce antisera directed exclusively against the carboxyl-terminal region of hPTH, which represents the predominant form of the hormone in blood; and 2) to determine whether a portion of the hormone other than the amino-terminal region has any of the biological activity of intact hormone or can bind to PTH receptors. The synthetic fragment was evaluated for heterogeneity by amino acid composition and by several high resolution analytical systems (thin layer electrophoresis, polyacrylamide gel isoelectric focusing, and Edman sequence analysis). Synthetic hPTH-(53-84) lacked PTH-like activity in both in vitro (renal) and in vivo (bone) assays and failed to inhibit the action of native PTH in vitro, indicating lack of receptor binding by the carboxyl-terminal region of the molecule. Because the fragment lacked biological activity, nonchemical evaluation was performed to confirm that the peptide represents the carboxyl terminus of hPTH. Immunological comparison of synthetic hPTH-(53-84) to native hPTH was undertaken using an RIA that employed an antibovine PTH antiserum whose antigenic recognition was limited to the carboxyl-terminal region of the hormone. Synthetic hPTH-(53-84) was more immunoreactive than native hPTH. The immunological findings provide further evidence that the synthetic peptide accurately represents the carboxyl-terminal region of native hPTH.


Renin release by parathyroid hormone in the dog. Synthetic N-terminal 34 amino acid peptide of bovine parathyroid hormone (PTH) produced a consistent rise in plasma renin activity in saline-loaded dogs when given iv either as a single bolus dose of 400 U or infused steadily in lower dosage over a number of hours. Infusion of as little as 1 U/min produced a significant rise in plasma renin activity, but a greater effect was obtained with 2 U/min. Infusion of 4 U/min had no more effect than 2 U/min. In contrast to transient hypotension after rapid injection of a single large dose, blood pressure did not change significantly during the steady infusion of lower doses of PTH. These findings are compatible with the thesis that an increased distal tubular load of sodium stimulates renin release and suggest that the increased distal sodium absorption after the proximal effects of PTH may be regulated by the renin-angiotensin-aldosterone system.


The metabolic clearance rate and uterine metabolism and retention of progesterone and 20alpha-hydroxypregn-4-en-3-one during the secretion of uteroglobin in ovariectomized, steroid-treated rabbits. The metabolic clearance rate (MCR) and in vivo uterine metabolism and retention of progesterone (P) and 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OHP) during the uterine synthesis of uteroglobin in the rabbit have been studied. Oophorectomized rabbits with the uteri ligated received daily im injections of oil vehicle (C), 2 mg P/kg BW, or 2 microgram estradiol/kg BW. The MCR of [14C]P and [3H]20 alpha-OHP did not differ among treatment groups and was 260 +/- 30 and 270 +/- 33(SE) liters/day, respectively. Although the uterine tissue to arterial blood ratio of [14C]P differed between groups (E greater than C greater than P), the tissue to blood ratio of 20 alpha-OHP was the same for the three groups and was about 1. The uterine extraction of P was 32 +/- 6.2(SE)% and was the same for the three treatment groups. The uterine extraction of 20 alpha-OHP was significantly increased by estrogen treatment [33.4 +/- 6.0(SE)%] compared to the control [14.8 +/- 3.3(SE%)] or P[16.9 +/- 4.5(SE)%] groups. These studies indicate that the ability of exogenous 20alpha-OHP to stimulate uteroglobin secretion is probably mediated by its peripheral conversion to P. These studies also indicate the value of continuous infusion of radioactive steroids to evaluate tissue retention and metabolism in vivo.


Plasma neurophysin and vasopressin in the rat: response to adrenalectomy and steroid replacement. Impaired excretion of a water load is known to occur in adrenal insufficiency and to be corrected by administration of glucocorticoid. Such impairment has been related to either a loss of a permissive effect of glucocorticoids on the diluting segments of the nephron or to an alteration of release, turnover, or action of antidiuretic hormone. Specific and sensitive RIAs for arginine vasopressin and neurophysin were utilized to measure plasma and pituitary levels of neurohypophyseal peptides at baseline and after an intragastrically administered water load. Conscious, unanesthetized, and nonstressed sham-operated, adrenalectomized, and adrenalectomized prednisone-treated rats were studied. The results demonstrate a significant elevation in vasopressin and neurophysin in plasma in adrenalectomized rats maintained in a normal state of hydration. After water loading, the adrenalectomized rats diluted their plasma osmolality but had a decreased urinary volume, increased urinary osmolality, and elevated vasopressin and neurophysin in their plasma. In the pituitary, vasopressin and neurophysin were depleted in adrenalectomized rats, indicating increased secretion of these peptides. It is concluded that elevated vasopressin in plasma may be an important factor in the incomplete water diuresis in adrenal insufficiency.


Hypothalamic and raphe serotonergic systems in ovulation control. The role of hypothalamic and midbrain serotonergic systems in the control of ovulation in the rat has been investigated using 5,7-dihydroxytryptamine (5,7-DHT) lesioning in conjunction with desmethylimipramine pretreatment. Intracisternal injection of 5,7-DHT (100 or 200 microgram) produced a dose-dependent decrease in the incidence of ovulation induced by PMS gonadotrophin (PMSG; 8 IU sc on day 30). Intraventricular injection of 200 microgram 5.7-DHT via the lateral ventricles completely blocked PMSG-induced ovulation. After injection into either site, nonovulatory animals were in proestrus on day 33 and contained fluid-filled uteri. Intracerebral injection of 5,7-DHT into the dorsal or median raphe significantly decreased the numbers of animals induced to ovulate. The extent and specificity of these chemical lesions were evaluated in the suprachiasmatic (SNR) and arcuate-median eminence regions of PMSG-treated rats using an in vitro uptake model. A general feature of every case of inhibited ovulation was the significant decrease in uptake of serotonin in the SNR, indicating destruction of serotonergic inputs to this region. This suggests that serotonergic SNR input from the dorsal raphe region is essential to ovulation. Median raphe lesions appeared to be more extensive than dorsal raphe lesions, involving serotonergic projections to the arcuate-median eminence region. In addition, ascending noradrenergic projections to the SNR were significantly destroyed, also implicating these systems in ovulation control.


Secretion of prostatic binding protein by rat ventral prostate: influence of age and androgen. Rat ventral prostate of adult male rats contain a large amount of prostatic binding protein (PBP). Immunological evidence indicates that this protein is a specific secretion product of this gland. The amount and concentration of PBP in ventral prostate show marked changes as a function of age. PBP is low but detectable (0.009 and 0.002 U/mg prostate) in 5- and 10-day-old rats and increases thereafter in a biphasic way to adult levels (0.619 U/mg prostate). After castration of PBP drops to 0.054 U/mg prostate after 10 days and 0.030 U/mg prostate after 21 days. The concentration of PBP returns to precastration levels after 2 weeks of androgen treatment. Estradiol and progesterone are ineffective in this respect. The antiandrogen, cyproterone acetate, counteracts the stimulatory effect of testosterone propionate.


Daily rhythmicity of serum testosterone concentration in the male laboratory rat. Daily rhythmicity of serum testosterone concentration in the mature male laboratory rat was examined under various lighting schedules. In rats living in a standard light cycle (12-h light, 12-h dark; lights on at 0600 h), a trimodal rhythm was predominant, with elevations near 0200, 1200, and 1800 h. This pattern was reasonably stable in seven different studies, despite differences in experimental design, method of blood collection, anesthesia, and whether individual rats were sampled once or repeatedly, and was found both in groups of animals and in individuals, including a study using 40-day-old rats. In constant illumination, the pattern was disrupted, but in constant darkness the trimodal pattern was maintained, indicating that the rhythm is endogenous. In a reversed light cycle (12-h dark, 12-h light; lights on at 1800 h), the "midday" elevation was reversed; in an altered light cycle (12-h dark, 12-h light; lights on at 2300 h), the time of the "midday" elevation was shifted. Serum testosterone concentration was higher during the light phase than the dark phase, and was higher in constant light than in constant darkness. A seasonal shift in the daily rhythmicity of serum testosterone concentration is suggested. The trimodal rhythmicity contrasts with the circadian rhythmicity of other hormones. Its functional role in the life of the animal is unknown.


Calcium-induced release of 5-hydroxytryptamine from thyroid lobes in vitro and accompanying ultrastructural changes in parafollicular and follicular cells. Biogenic amines, including serotonin (5-HT), have been shown to activate follicular cells of the thyroid. 5-HT is stored in bat's parafollicular cells. Previous radioautographical evidence indicates that this 5-HT is present in calcitonin granules. The present study was done to determine if Ca++, the natural stimulus to calcitonin release, would also release parafollicular cell 5-HT and, if so, whether this release would be accompanied by activation of follicular cells. Parafollicular cells were filled with labeled 5-HT by incubation of thyroid lobes of bats or mice with 5-[3H]hydroxytryptophan, the precursor of [3H]5-HT. Thyroid lobes were incubated in vitro in Ca++-free medium containing a chelating agent and were then challenged with Ca++ (0-30 mM). Release of [3H]5-HT was defected beginning at 5 mM Ca++. [3H]5-HT release was roughly proportional to the Ca++ concentration. Calcium challenge also affected thyroid ultrastructure in bats. After 10-min exposure to 30 mM Ca++, there were an increased number and centripetal movement of follicular cell lysosomes, development of apical pseudopods, and formation of colloid droplets. Many lysosomes also developed a crystalline-like matrix. An unique membrane-enclosed, rod-shaped organelle appeared in a small number of follicular cells. Parafollicular cells exposed to high concentrations of Ca++ were often degranulated, although many appeared unchanged. These changes confirm that 5-HT is released from parafollicular cells by elevating the external Ca++ concentration, supporting the hypothesis that 5-HT and calcitonin share storage granules. 5-HT release may mediate the activation of follicular cells by Ca++.


Serotonin receptor antagonists block a natural, short term surge in serum growth hormone levels. Serum GH levels in untreated rats were found to increase spontaneously to over 300 ng/ml around the onset of darkness (1900 h) and then decrease to under 100 ng/ml by 2000 h. Pretreatment with the serotonin receptor blockers, metergoline (0.2, 0.5, 1, or 2 mg/kg) or cyproheptadine (1, 2, or 5 mg/kg), at 1800 h significantly blunted this physiological rise in serum GH. In rats bearing chronic right atrial cannulae from which blood samples were drawn every 15 min for 5 h, the administration of metergoline (1 mg/kg) was also observed to reduce plasma GH levels over a 3.25-h period. Serotonin-containing neurons in the brain thus seem to be involved in the physiological secretion of GH.


Regulation of the uterine response to estrogen by thyroid hormone. After estradiol (E2) administration, early increases (within 4 h) in uterine wet weight and the synthesis of 2-deoxyglucose-6-phosphate from 2-deoxyglucose are similar in ovariectomized rats and in ovariectomized rats made hypothyroid by feeding a low iodine diet containing prophylthiouracil. Most late uterine responses occurring 24 h after E2 treatment, however, are greatly diminished in the hypothyroid animals. The diminished responses include increases in uterine wet weight, dry weight, protein content, RNA content, and the incorporation of thymidine into uterine DNA. One response, the synthesis of 2-deoxyglucose-6-phosphate from 2-deoxyglucose is not diminished in hypothyroid rats 24 h after E2 treatment. The diminished uterine response is not due to a shift in the dose-response curve for E2, but results from a decrease in the magnitude of the maximum uterine response. Treatment of hypothyroid rats with exogenous T3 restores the diminished uterine response in a dose-dependent and time-dependent manner. A dose of 0.5 microgram T3/100 g BW for 5 days restores the response completely, while 48-72 h of treatment with higher doses of T3 are required to restore the response. The effect of T3 is not mediated by the pituitary, since exogenous T3 restores diminished uterine responses in ovariectomized, hypophysectomized animals. These results suggest that thyroid hormones may have a direct effect on the uterus which regulates the responsiveness of the organ to E2.


L-Leucine-induced secretion of glucagon and insulin, and the "off-response" to L-leucine in vitro. I. Characterization of the dynamics of secretion. The effects of L-leucine, D-leucine, and L-isoleucine upon the secretion of glucagon and insulin were investigated using the isolated, perfused rat pancreas. All experiments were conducted in the presence of 5.6 mM D-glucose. Ten-minute perfusions of 2, 5, and 10 mM L-leucine induced the release of glucagon and insulin in a dose-related manner. The removal of L-leucine was followed by renewed release of insulin ("off-response") but not of glucagon. The magnitude of the off-response was greater when L-leucine was perfused over longer periods. L-Isoleucine evoked the release of both glucagon and insulin. When L-leucine was administered during perfusion of L-isoleucine, L-leucine-induced release of glucagon was inhibited, that of insulin was augmented, and the insulin off-response prevailed. When the perfusion of L-leucine immediately preceded that of L-isoleucine, L-isoleucine-induced release of glucagon was abolished and that of insulin was augmented. D-Leucine evoked the release of glucagon but not of insulin, and no off-response occurred. When the perfusion of D-leucine followed that of L-leucine, D-leucine-induced glucagon release was inhibited; the insulin off-response to L-leucine was not altered. We reached the following conclusions. 1) Glucagon release induced by L-leucine, D-leucine, or L-isoleucine is likely to be related to the occupancy by these analogous amino acids of transport and/or receptor sites which they share. 2) The insulin off response to L-leucine seems to be evoked by events which take place during the period of administration of L-leucine; these events are not likely to be the release of insulin that occurs during perfusion of L-leucine or the transport of L-leucine into or out of the beta cell. 3) Structurally or chemically similar compounds which are secretagogues both for glucagon and insulin affect the release of these hormones in different ways; these differences are likely to be due to dissimilar mechanisms governing the secretion of the two hormones.


Effect of streptozotocin-induced diabetes on hepatic ornithine decarboxylase activity in the rat. The effect of diabetes on rat hepatic ornithine decarboxylase (ODC) activity was studied in male rats 4 h to 8 days after the ip or iv administration of streptozotocin. Hepatic ODC activity increased 4-fold above control on the fourth day of diabetes. Increased ODC activity was observed with a dose of streptozotocin (70 mg/kg iv) which increased plasma ketones and glucagon and reduced plasma insulin. No change was seen with doses causing only mild hyperglycemia without change in plasma ketones, glucagon, or insulin. Insulin therapy prevented the increase in hepatic ODC activity in diabetic rats. Adrenalectomy or hypophysectomy also prevented the diabetes-associated increase of ODC activity. The studies suggest that insulin deficiency may result in increased hepatic polyamine synthesis.


Stimulation of steroid synthesis by normal rat adrenocortical cells in response to antimicrotubular agents. The role of cytoplasmic microtubules in the process of steroidogenesis was examined in enzyme-dispersed and mechanically dispersed adrenocortical cells obtained from adult male rats. These cells were maintained in culture for periods of 2 h to 2 days and tested with various antimicrotubular drugs as well as ACTH and N6,O2-dibutyryl adenosine 3',5'-monophosphoric acid [(Bu)2cAMP]. The drugs tested, viz. colchicine, vinblastine, and nocodazole, were found to stimulate steroid synthesis in a dose-dependent fashion; the maximal degree of stimulation was found to be the same as that obtained with either ACTH or (Bu)2cAMP. When a maximal concentration of an antimicrotubular drug was mixed with a maximal concentration of either (Bu)2cAMP or ACTH, no additive effect on steroidogenesis was observed. The mixture of maximal concentrations of two antimicrotubular drugs produced a steroidogenic response that was less than when either drug was used alone. However, when submaximal concentrations of two antimicrotubular drugs were mixed, a synergistic response was noted. These antimicrotubular drugs are known to interfere with microtubular function by binding to specific sites on the tubulin protomer and inhibiting normal polymerization. Lumicolchicine, the photoinactivated form of colchicine, does not bind to tubulin and did not stimulate steroid synthesis. These studies support the notion that microtubules can influence the process of steroidogenesis.


Blood-brain barrier restriction of peptides and the low uptake of enkephalins. Blood-brain barrier penetration of leucine-enkephalin, methionine-enkephalin, and other peptide-like compounds was measured after intracarotid injection of three isotopes and was found to be non-saturable over the nanomolar range of concentrations tested. No significant differences in brain regional extraction of leucine enkephalin (or morphine or heroin) were observed. In contrast to previous reports, the brain extraction of enkephalins was minimally low (E = 2-3%) and about the same order of magnitude as other putative neurotransmitters. Brain extractions of other peptide-like compounds were similarly small: TRH, E = 1%; glutathione, E = 0.5%; beta-alanyl histidine, E = 1%; and thioacetyl coenzyme A, E = 2%. Extraction of the non-diffusible reference dextran was determined to be 1%, suggesting that the blood brain barrier tends to restrict peptide penetration.


Progesterone metabolism by cultured Sertoli cells. The ability of Sertoli cells to metabolize progesterone to testosterone (T) and 5alpha-dihydrotestosterone (DHT) was investigated in vitro. Cultures of Sertoli cells isolated from testes of 80-day-old rats were incubated with [7(n)-3H]progesterone (10 muCi/3.0 nmol) for 0.5, 1, 2, and 3 h. The amount of progesterone converted to T, androstenedione (A), DHT, 17alpha-hydroxyprogesterone, and 20alpha-dihydroprogesterone was calculated on the basis of crystallization data. The amount of substrate converted to the various steroids increased between 0.5-3 h of incubation. During this period, T increased 6-fold; A, 2-fold; DHT, 4-fold; 17alpha-hydroxyprogesterone, 3-fold; and 20alpha-dihydroprogesterone, 12-fold. The amount of substrate converted to C-19 steroids (T, A, and DHT) increased linearly with time; 19.4 ng androgen/1 x 10(6) cells were formed within 3 h, the largest amount (11.83 ng) being DHT. Although the amount of 3H-labeled C-19 steroids formed from [3H]-progesterone by Sertoli cells is relatively small (1.8% conversion) compared to T formation by whole testicular tissue or by isolated interstitial cells, the ability of Sertoli cells to form T and DHT from progesterone may be physiologically important in the local regulation of Sertoli cell function and spermatogenesis.


Effects of chronic treatment of intact and hypophysectomized rats with thyroid-stimulating hormone on the metabolism of [35S]methimazole and [35S]propylthiouracil. Chronic treatment of intact rats with various doses of TSH increased the thyroidal 35S accumulation after single doses of [35S]methimazole (MMI) and [35S]propylthiouracil (PTU). However, no effect on the intrathyroidal breakdown of the drugs was observed. Thus absolute thyroidal levels of unmetabolized MMI and PTU were increased by factors of up to 2 and 3, respectively, compared to the control groups. Simultaneous decreases in the levels of thyroidal total iodine were observed. Hypophysectomized rats showed a marked inhibition of both thyroidal accumulation and oxidation of [35S]-MMI but TSH treatment of hypophysectomized rats restored the accumulation and oxidation to sham-operated and control group levels. The results show that in rats TSH has an important role in the control of thyroidal levels of antithyroid drugs currently used in the treatment of hyperthyroidism.


Regulation of phosphoenolpyruvate carboxykinase and fatty acid synthetase in brown fat of suckling rats. Infant rats were injected with prednisolone (0.5-5 mg/100 g wt). This caused phosphoenolpyruvate carboxykinase (PEPCK) activity to rise in liver and to decrease in brown fat. Fatty acid synthetase (FAS) activity remained unchanged in liver but increased in brown fat. A single injection of prednisolone caused hepatic PEPCK activity to remain elevated for at least 7 days. Brown fat FAS also remained high for that period. However, brown fat PEPK activity returned to normal on the third day after the injection. A single injection of prednisolone or cortisone to 5-day-old rats caused a transient elevation of the blood level of insulin and a prolonged decrease in that of growth hormone. No effect on the level of glucagon was noted. Injections of insulin had effects similar to those of prednisolone, i.e. a rise in hepatic and a fall in brown fat PEPCK. Using antibodies prepared to hepatic PEPCK it was shown that the observed changes were due to changes in the rate of synthesis of the enzyme. Using actinomycin D indirect evidence was obtained that changes in FAS activity of brown fat were also due to changes in the synthetic rate.


The effect of hypophysectomy on the quail's oviduct response to low and high doses of estradiol benzoate. The response of the quail's (Coturnix coturnix Japonica) oviduct to different doses of estradiol benzoate (EB) was explored in ovariectomized and ovariectomized-hypophysectomized animals. Doses of EB ranged from 10 microgram to 1 mg/animal daily for 6 consecutive days; animals were sacrificed 24 h after the last injection. Controls were injected with olive oil. Parameters thought to reflect stimulation of the oviduct were 1) wet and dry weight, 2) DNA, 3) RNA, 4) soluble proteins, 5) ovalbumin, and 6) morphological changes at the level of magnum's mucosa. A statistical significant difference in several of the parameters listed was found between the effect of different doses of EB and whether the injection was done in ovariectomized or in ovariectomized-hypophysectomized animals. The dose response with EB for cell proliferation and induced proteins accumulation (ovalbumin) show that it takes considerably more EB to maximally stimulate ovalbumin accumulation when compared with cell proliferation. Hypophysectomy decreases the oviduct responses to high EB doses. Ultrastructural studies of the magnum mucosa confirm these biochemical results. The evidence presented suggesting an EB dose-dependent difference induction of specific cellular responses in the presence and absence of the pituitary in quails are the basis to postulate the contributing role of the pituitary in the parameters measured.


The effect of zinc on insulin metabolism. Experiments were designed to study the effect of Zn on in vivo and in vitro insulin metabolism. The in vivo experiments involved pretreating mice with either Zn or Na, followed by ip [125I]iodoinsulin injection. Pretreatment of mice with Zn resulted in an accelerated and increased magnitude of binding of [125I]iodoinsulin to the liver compared to mice pretreated with Na. Results are submitted which support the probability that the changes in the amounts of intact and degraded insulin in circulation with time are related to the binding and degradation of insulin in the liver rather than in the kidney. In vivo ip injected insulin was demonstrated to preferentially bind to the plasma membrane of the liver. Liver plasma membranes isolated from mice pretreated with Zn bound more [125I]iodoinsulin than plasma membranes of Na-pretreated mice. In vitro experiments employing isolated liver plasma membranes demonstrated that added Zn increased the binding and inhibited the degradation of insulin. Evidence is presented that supports the concept that two receptors exist, one at which degradation of [125I]iodoinsulin occurs and another at which degradation does not occur.


New mineralocorticoids: 5alpha-dihydroaldosterone and 5alpha-dihydro-11-deoxycorticosterone. Mineralocorticoid activity of several delta4-3-ketosteroids and their 5alpha-dihydro analogs were evaluated by bioassay using urinary Na:K ratio of adrenalectomized rats as an index of mineralocorticoid activity. Among delta4-3-ketosteroids, aldosterone, 11-deoxycorticosterone, corticosterone, cortisol, 11-dehydrocorticosterone, and cortisone showed mineralcorticoid activity with aldosterone, the most potent of the series, showing virtually maximum activity at a dose of 0.25 microgram/rat. 5alpha-Dihydroaldosterone and 5alpha-dihydro-11-deoxycorticosterone possessed distinct mineralcorticoid activity, albeit less than aldosterone and 11-deoxycorticosterone. 5alpha-Dihydrocorticosterone, 5alpha-dihydrocortisol, 5alpha-dihydro-11-dehydrocorticosterone, and 5alpha-dihydrocortisone did not show mineralocorticoid activity in doses up to 100 microgram/rat. It is concluded that reduction of the 4.5 double bond diminishes mineralocorticoid activity of delta4-3-ketosteroids. Nevertheless, 5alpha-dihydroaldosterone has distinct mineralocorticoid activity with potency of 1.8% of that of aldosterone and approximately the same as that of 11-deoxycorticosterone.


Effect of parturition on serum iodothyronine concentrations in fetal sheep. T4, T3, and rT3 were determined by RIA on daily serum samples collected from five catheterized fetal sheep during the 2-week period before spontaneous vaginal delivery (PTD). Samples were also available from three newborn lambs during the first 4 h of life. T3 concentrations began to increase 4-6 days before spontaneous vaginal delivery, from a stable geometric mean concentration of 30 ng/dl to a mean concentration of 131 ng/dl on the day of delivery (day 0). T4 concentrations decreased in four of the animals and remained unchanged in one during this time interval. rT3 concentrations decreased from a stable mean of 472 to 249 ng/dl on day 0. The prenatal increase in serum T3 levels correlated temporally with increasing fetal serum cortisol concentrations. The mean T3 concentration increased further after delivery to a level of 487 ng/dl at 1 h of age without significant changes in T4 or rT3 levels. The following conclusions were reached: 1) fetal serum T3 concentrations increase while T4 and rT3 concentrations decrease during the 4-6 days preceding spontaneous labor, and 2) T3 concentrations increase further after delivery. Available evidence suggests that the prenatal increase in fetal serum T3 levels is mediated by a cortisol-induced increase in T4 to T3 conversion in fetal tissues. The mechanism(s) of the postnatal increase in serum T3 concentrations in the newborn is not clear.


[des-Asp1]angiotensin II in the dog: blood levels and effect on aldosterone. Circulating levels of [des-Asp1]angiotensin II ([des-Asp1]-AII), angiotensin II (AII), and aldosterone were measured in five conscious beagle dogs before and during iv infusion of [des-Asp1]AII at rates of 3, 6, 12, and 24 ng/kg/min. The animals were studied after 4 days on a normal sodium and potassium diet and again after a period of sodium depletion accomplished by iv furosemide (2-5 mg/kg) and 4 days of low sodium diet (2-5 mmol/day). Compared to the normal sodium diet, sodium depletion resulted in increases in the plasma levels of aldosterone from 10 +/- 2 (SE) to 66 (16-116) ng/100 ml of AII from 16 +/- 4 to 52 +/- 13 pmol/liter and of [des-Asp1]AII from 2 +/- 0.7 to 12 +/- 4 nmol/liter. Incremental infusions of [des-Asp1]AII in the sodium replete state resulted in progressive increases in the plasma levels of aldosterone in all dogs. In comparison with a previous study in which dogs were infused with AII, it was apparent that [des-Asp1]AII was equally or slightly more potent in stimulating aldosterone and had a higher metabolic clearance rate than AII. [des-Asp1]AII stimulated aldosterone in four of the five sodium-depleted dogs but no steepening of the [des-Asp1]AII/aldosterone dose-response curves was apparent. These results do not support the hypothesis that circulating [des-Asp1]AII mediates the effect of AII on aldosterone in the dog.


A suspension culture of pinealocytes: regulation of N-acetyltransferase activity. A method to prepare suspension cultures of highly viable pinealocytes is described. Pineal glands of neonatal or adult rats can be used; the best yield is obtained from neonatal glands. Morphological examination of the cells indicates that they have a normal, healthy appearance and retain some structures typical of neonatal pinealocytes. During the first 24 h of culture, the cells aggregate into small clusters; after several days, larger aggregates can be seen. Biochemical studies indicate that adrenergic stimulation of these cells causes a 30- to 100-fold increase in serotonin N-acetyltransferase activity, a response which is unique to the pineal gland. The relative order of potency of several adrenergic agonists is l-isoproterenol greater than l-norepinephrine greater than or equal to l-epinephrine greater than phenylephrine. Serotonin, tyramine, histamine, carbachol, gamma-aminobutyric acid, cocaine, and desmethylimipramine are inactive. Studies using adrenergic and metabolic inhibitors indicate that the regulation of N-acetyltransferase in these cells has the same characteristics of that seen in adult tissue. In addition, these cells synthesize protein and RNA from radioactive precursors, convert tryptophan to serotonin, and N-acetylate endogenous serotonin.


Angiotensin II concentration in cerebrospinal fluid after intraventricular injection of angiotensinogen or renin. To determine if the brain enzyme which has renin-like activity in vitro can form angiotensin in vivo, angiotensin II concentration in cerebrospinal fluid (CSF) was measured before and at various intervals after injection of partially purified renin substrate (angiotensinogen) into the third cerebral ventricle of anesthetized dogs. The injection increased CSF angiotensinogen concentration 3-fold, but despite this, CSF angiotensin II concentration, which was undetectable (less than 6.25 fmol/ml) before injection, did not change. Arterial blood pressure was also unchanged after the injection. In contrast, both CSF angiotensin II concentration and arterial pressure increased after an inventricular injection of renin. These results demonstrate that angiotensin II is formed centrally after administration of exogenous renin but not after injection of angiotensinogen. The results thus fail to demonstrate renin activity in the brain in vivo.


Intranuclear binding of [3H]dihydrotestosterone by cultured human fibroblasts. The uptake of [1,2,4,5,6,7-3H]dihydrotestosterone into whole cells and nuclei has been assessed in fibroblasts grown from genital skin of 10 controls and 9 subjects with hereditary male pseudohermaphroditism due to androgen resistance. The cells were exposed to hypotonic buffer and ruptured by passage through a 25-gauge needle, and the nuclei were purified by sedimentation through 2.1 M sucrose. Uptake of the hormone into nuclei reached an apparent plateau in 45 min, was saturable at 1 nM dihydrotestosterone, and was not detectable in the presence of excess nonradioactive hormones. Over a wide range of uptake by intact cells from control subjects and from subjects with androgen resistance due to 5alpha-reductase deficiency or receptor deficiency, nuclear uptake averaged about half of the total cell uptake. Furthermore, in cells from two unrelated 46,XY phenotypic females with androgen resistance but normal 5alpha-reductase activity and normal whole cell dihydrotestosterone binding, uptake into the nucleus was also normal. In these subjects, the defect in androgen action must be at some terminal phase of androgen action.


Early in vitro induction of rat pituitary GH mRNA by T31. Previous work has shown that thyroid hormone stimulates rat pituitary GH synthesis and GH mRNA activity and concentration. However, the earliest demonstration of increase in GH mRNA activity was 24 hours following T3 addition whereas stimulation of GH synthesis has been observed 2 hours after treatment with T3. Thus, it is unknown whether increase in pituitary GH mRNA is a prerequisite for the stimulation of GH synthesis. In the present investigation in vitro addition of 1.5 x 10(-10) M T3 to pituitaries isolated from hypothyroid rats resulted in a slight but significant increase of GH mRNA activity within 2 hours. Further stimulation of GH mRNA activity was observed over the period of 12 hours. No increase of GH mRNA activity occurred in the absence of T3, and T3 had no effect on the PRL mRNA activity. These findings suggest that increase in GH mRNA may be responsible for the observed induction of GH synthesis, and that at least one of the primary actions of thyroid hormone is at the nuclear level.


Lactoferrin is a marker for prolactin response in mouse mammary explants. We recorded a bioassay for prolactin using densitometric analysis on polyacrylamide gels of caseins produced by mammary explants in vitro. A protein which also increased in the presence of increasing amounts of prolactin in the medium has now been identified. Its migration in several electrophoretic systems is the same as that of a whey protein of mouse milk, lactoferrin. It is an iron-binding glycoprotein with a molecular weight by SDS gel electrophoresis equal to mouse, human and bovine lactoferrin. It gives a precipitin reaction in an immunodiffusion system with monospecific antiserum raised in rabbits against mouse lactoferrin.


19-Nor deoxycorticosterone (19-nor DOC): mineralocorticoid receptor affinity higher than aldosterone, electrolyte activity lower. By screening urine extracts from rats with adrenal regeneration hypertension, Gomez-Sanchez et al. found a steroid, subsequently identified as 19-nor DOC, with high affinity for tritiated aldosterone (3HA) binding sites in rat kidney cytosol. We here report studies on the affinity of authentic 19-nor DOC for mineralocorticoid receptors, its binding in plasma and its activity in the rat urinary mineralocorticoid assay. When kidney slices from adrenalectomized rats were incubated in protein-free buffer with 3HA, 19-nor DOC consistently competed better (approximately 140%) for 3HA binding sites than did equivalent concentrations of non-radioactive aldosterone. Under identical conditions, save for the inclusion of 20% adrenalectomized rat plasma in the incubation medium, 19-nor DOC shows only approximately 40% the potency of aldosterone in displacing 3HA. Determination of renal binding of 3HA after injection of 3HA +/- aldosterone +/- 19-nor DOC in vivo similarly shows 19-nor DOC to be approximately one third as potent a competitor for 3HA binding sites as aldosterone. In the rat urinary bioassay, 19-nor DOC shows no antagonist activity when injected with aldosterone; in the absence of aldosterone, 19-nor DOC acts as a mineralocorticoid agonist, with an apparent potency 10-30% that of aldosterone. Conclusions of the study are therefore (i) at a molecular level, 19-nor DOC has a higher affinity than aldosterone for mineralocorticoid receptors, (ii) in vivo, its potency in terms of receptor occupancy is markedly lower than that of aldosterone, due to higher levels of plasma binding, (iii) in effector terms, 19-nor DOC is a full agonist without antagonist activity.


A newly designed method for removal of the pineal body, and depression of convulsions and enhancement of exploratory movements by pinealectomy in mice. We designed a simple technique for surgical excision of the pineal body in mice. Such can be accomplished in 10 min by a skilled worker and there are no lesions whatever. In pinealectomized El-strain mice the appearance of convulsions was inhibited up to 30 days, while intact El-mice were all fallen into convulsions by being shaken up and down on a flat carton. Pentazole-induced convulsions propagate from one dd-mouse to another in an aggregated state, but such propagation did not occur in pinealectomized animals. Exploratory movements of pinealectomized dd-mice increased as compared to intact animals and the increase was always more extensively observed in El-mice in which, even in the intact animals, the exploratory movements were much more than those of dd-mice. From the above-mentioned experiment, it is considered that the pineal body plays a role not only in initiating but also in propagating convulsions, and that it may have a depressive action on motor activities in mice.


Competitive protein binding assay for 1,25-dihydroxy-vitamin D in human plasma. A sensitive and simplified radioreceptor assay for 1, 25-dihydroxyvitamin D (1, 25-(OH)2D) in human plasma was described and applied to preliminary clinical studies. Tritium-labeled 1, 25-(OH)2D3 was produced by incubating chick kidney homogenate with tritium labeled 25-hydroxyvitamin D3 (25-OHD3). A cytosol receptor was obtained from rachitic chick intestine (Kd=5.3 X 10(-11) M). Lipids in 5 ml of heparinized human plasma were extracted with dichloromethane, and 1, 25-(OH)2D was isolated by a Sephadex LH-20 column followed by high pressure liquid column chromatography. Recovery of 1, 25-(OH)2D3 after the plasma extraction and chromatography ranged from 58 to 100%. The assay was sensitive to 5 pg/tube. Diluted plasma from a patient on a high dose of 1 alpha-OHD3 showed a dilution curve parallel to the standard curve. The cytosol receptor showed a cross reactivity to various vitamin D3 metabolites physiologically present in the circulation and it was thought to be essential to eliminate other vitamin D3 metabolites 1,25-(OH)2D from plasma samples by high pressure liquid chromatography. Plasma concentrations of 1, 25-(OH)2D were, in the case of most normal subjects, distributed from 7 to 33 pg/ml and the range of distribution became greater in relation to age, indicating that plasma values should be matched to age. Whereas markedly high values of 1, 25-(OH)2D in plasma were found in some cases of primary hyperparathyroidism with prominent bone resorption, relatively low values were seen in some patients with chronic renal failure, senile osteoporosis, osteomalacia and hypercalcemia due to bone metastasis.


Re-evaluation of exercise as a screening test for ruling out human growth hormone deficiency. Human growth hormone (HGH) response to exercise was studied in 27 normal children and 152 children with a chief complaint of short stature and in 9 patients with HGH deficiency. Of the 152 children 107 could be ruled out of HGH deficiency since HGH level increased to more than 5 ng/ml during exercise, and of the remaining 45 children of non-responders to exercise, 22 were ruled out of HGH deficiency since bone age to chronological age ratio was more than 0.9 and/or height increment for the past 3 years was more than 15 cm. Seventeen patients were hospitalized for further examinations, which resulted in finding normal HGH secretion in response to insulin-induced hypoglycemia, arginine infusion and/or propranolol-glucagon test in 12 children and also in finding no HGH deficiency in children higher than -2.0 S.D., but 5 HGH-deficient children in those lower than -2.0 S.D. Exercise can be used as a screening test for ruling out HGH deficiency when HGH secretion in response to exericse is evaluated by height, bone age and height increment of the patients.


Comparison of two angiotensin II analogues in normal subjects and hypertensive patients. Two angiotensin II analogues, i.e., 1-sarcosine, 8-isoleucine angiotensin II (Sar1, Ile8-AII) and 1-sarcosine, 8-alanine angiotensin II (Sar1, Ala8-AII), are now available in the clinical study. Comparative studies of the antagonistic potency and the agonistic effect of these two AII analogues were made in normal subjects on three sodium balances and in hypertensive patients with various etiologies on sodium depletion. Both AII analogues had an agonistic pressor effect in normal subjects. This effect changed with different sodium balances. In the low sodium state, this agonistic action was minimized. The agonistic pressor effect of Sar1, Ile8-AII was greater than that of Sar1, Ala8-AII in all sodium states. There was found an agonistic activity of both AII analogues not only on blood pressure, but also on renin and aldosterone secretion, and renal vasculature in normal subjects on a regular diet. The antagonistic depressor potency of both compounds was also varied by changing sodium balance, being greatest in the low sodium state. In hypertensive patients on sodium depletion, the blood pressure responses of individual patients to these two AII analogues were significantly correlated (r = 0.8, n = 20). These results indicate taht pretreatment of sodium depletion is necessary to prevent the side effect caused by the agonistic pressor action of AII analogue, and also to predict renin depency in hypertensive patients efficiently.


Effect of secretin on plasma insulin and glucagon in man. To investigate the effect of physiological doses of secretin on pancreatic A and B cell functions, secretin was infused at a rate of 0.5, 1.0 and 2.0 clinical unit per kg body weight per hr into eight normal men for 30 min or 60 min after an over-night fast, and changes in plasma immunoreactive insulin (IRI) and glucagon immunoreactivity (GI) were measured while monitoring circulating secretin levels by a radioimmunoassay. During the infusion of secretin, the plasma immunoreactive secretin (IRS) levels rose to 140-390 pg/ml which was within a range of the physiological fluctuation in plasma secretin levels reported hitherto. No significant alteration of plasma IRI and GI levels could be demonstrated during these simulated physiologic infusions of secretin. These data suggest that, in humans, the physiological dose of secretin does not influence insulin and glucagon secretion from the pancreas in the basal state.


Prolactin binding in pregnancy-dependent mammary tumors in GR/A mice. Prolactin binding in normal mammary glands and pregnancy-dependent mammary tumors and plasma prolactin levels were assayed near parturition in GR/A female mice. Mammary glands and tumors cut into 0.5-mm thick slices were incubated in Medium 199 containing 5 ng of 125I-labeled ovine prolactin at 37 degrees C for 2 hr, and specific prolactin binding was expressed in terms of the difference between total and nonspecific bindings per mg tissue. On day 19 of pregnancy, prolactin binding was low in both normal mammary glands and tumors. One day after parturition, the binding of nonlactating mammary gland was as low as that on day 19 of pregnancy. On the other hand, the binding of lactating glands was raised significantly. There was no difference in the binding of tumors between lactating and nonlactating animals and both were significantly higher on day 1 after parturition than on day 19 of pregnancy. The plasma prolactin level assayed by radioimmunoassay was significantly higher on day 19 of pregnancy than on day 1 after parturition regardless of lactation. No relations were observed between the plasma prolactin level and the prolactin binding of either normal or neoplastic mammary glands.


The suggestive roles of the anterior hypothalamic area on prolactin  secretion in hemiovariectomized and ovariectomized rats. Serum prolactin concentration was studied in hemiovariectomized rats, hemiovariectomized rats bearing chronic estradiol benzoate (E2) implant either in the anterior (A-AHA) or in the middle (M-AHA) portion of the anterior hypothalamic area (AHA), and was also studied in ovariectomized AHA lesioned rats. Hemiovariectomy caused rise of serum porlactin which peaked at day 10 and thereafter declined to the basal level. E2 implanted in the M-AHA either unilaterally or bilaterally temporarily suppressed the serum porlactin rise induced by hemiovariectomy. Unilateral implant of E2 in the A-AHA elicited also a temporary but longer suppressive effect. Bilateral E2 implant in the A-AHA consistently suppressed serum prolactin of hemiovariectomized animals. Ovariectomized AHA lesioned rats responded to systemic E2 treatment with a slight increase of serum prolactin while the non-lesioned ovariectomized rats responded with a significant increase. The additional E2 treatment effectively enhanced serum prolactin elevation in AHA ablated animals. The results substantiated the involvement of the AHA in prolactin secretion as well as they disclosed the A-AHA as a dominant area while the M-AHA appears to be supplementary area for prolactin inhibition, provided that the areas were in response to E2 implantation.


Hypothalamo-hypophyseal-gonadal system in male rats and guinea pigs treated with the antiandrogen 4-nitro-3-trifluoromethylisobutyranilide. Influence of the competitive testosterone antagonist 4-nitro-3-trifluoromethylisobutyranilide (NFBA) on the hypothalamo-hypophyseal-gonadal system in male Wistar rats and guinea pigs was studied. NFBA as well as nonlabelled testosterone diminished 3H-testosterone uptake in hypothalamus and hypophysis of castrated animals. Increase of plasma testosterone level was obtained after the antiandrogen treatment in a dose of 10 or 25 mg/kg per os during 3, 10 and 48 days. Maximal level of the hormone was noticed on the 10th day. The contents of 17-ketosteroids in testes had not changed, but the testosterone content and steroid-delta5-3beta-ol-dehydrogenase activity were high during the treatment period. After 10 days of NFBA administration increased plasma LH level was found. Increase in size and number of gonadotropocytes was revealed by histological and histochemical methods as well as Leydig cell hypertrophy. High plasma testosterone level and Leydig cell activation were obtained in guinea pigs as a result of NFBA subcutaneous injections in a dose of 10 mg/kg during 30 days. The suggestion is made that prolonged NFBA treatment produces changes in the testosterone metabolism. Short-term administration of NFBA may be used for the estimation of the hypothalamo-hypophyseal functional abilities.


Effect of long-term treadmill running on gonadal activity in female rats. Histochemical studies of delta5-3beta-HSD and G-6-PD activities in the ovaries of female rats undergoing long term-treadmill running showed a marked reduction. This change was associated with increased concentration of total cholesterol and total ascorbic acid in the same organ. All these results suggest ovarian hypoactivity.


Peripheral conversion and uptake of androgens in a XXY-man with Klinefelter's syndrome. The conversion of 3H-testosterone and the uptake of 3H-testosterone and 3H-5alpha-dihydrotestosterone were investigated in pubic skin and pubic hair follicles of a XXY-man with inadequate pubic hair. The uptake of both androgens was demonstrated in the skin as well as in the hair follicles. Similarly the activity of steroid 5alpha-reductase was present in both tissues. The total conversion of 3H-testosterone was 2-3 times higher in the patient than in control persons. In the XXY-man the major metabolites were 5alpha- and 5beta-androstanediols, whereas in the normal men 5alpha-dihydrotestosterone and 4-androstenedione were mainly formed from testosterone. An explanation of the inadequate growth of pubic hair in our patient seems to be related to a conversion of testosterone - at its low plasma level - to its relatively inactive metabolites.


Androgen metabolism in the epithelium of the bovine cornea. The metabolism of [14C]-testosterone and [14C]-androstenedione resp. was investigated in the epithelium of the cow cornea. [14C]-testosterone was converted mainly to androstenedione (48.3%), while conversion to dihydrotestosterone accounted only for 2%. On incubation of androstenedione mainly testosterone was formed (9.8%) and polar, probably hydroxylated, substances (7.6%).


Posterior pituitary dopamine and noradrenaline content: effect of thirst, ethanol and saline load. Dopamine and noradrenaline content in the posterior pituitary were found to be lower in alcohol-loaded unanesthetized rats than in rats thirsting for 72 h. Dopamine content was also found to be lower in anesthetized saline loaded rats in comparison with hydropenic animals. It is concluded that in situations in which an increase of vasopressin release is expected, the posterior pituitary content of catecholamines will also be increased, and vice versa, which might imply their role in the release of vasopressin.


A simple and rapid technic for radioimmunoassay and calculation of plasma steroid concentrations. A simple RIA technic for plasma steroids is described: Extraction residues or reference standards are dissolved in 20 microliter ethyl alcohol, or 20 microliter of alcoholic plasma extracts are given directly into RIA tubes; 500 microliter tracer solution, containing a known tracer mass, and, after mixing, 500 microliter antiserum solution or buffer solution are added, followed by 20 min incubation at room temperature and 20 min standing in an ice bath.  Free from bound activity is separated by charcoal coated florisil or silicagel. Plasma steroid levels are calculated by a simple equation. An immunologic equivalent to the tracer mass present in the reaction mixture serves as reference value. Drawing of standard curves is not necessary for every assay. The linear transformation used here allows the number of reference standards to be reduced. So also single samples can be analyzed with relatively low effort.


Colloid resorption in thyroid glands as revealed by scanning electron microscopy. Colloid resorption from thyroid follicles in control and in calcitonin-treated rabbits was studied with the scanning electron microscope. Thus, an intense cell surface activity with several vesicles (exocytotic vesicles) and large pseudopods, sometimes cylindroids, is predominant on the thyroid follicular cells of calcitonin-treated rabbits. The pseudopods are irregular, mostly elongated, cup or dome-shaped and operate by phagocytosis or engulfment of colloid droplets (endocytosis). Only microvilli are visible on the cell surface of thyroid follicles in control rabbits. Comparative ultrastructural studies also revealed the occurrence of small exocytotic vesicles and large pseudopods in thyroid glands stimulated by calcitonin. The present findings demonstrate that the scanning electron microscopy better visualizes the colloid resorption and provides more details for the role of follicular cell surface in thyroid physiology.


Inactivation of endogenous FSH by neuraminidase from Vibrio cholerae. Counting experiments on ovary-slices of animals treated with Neuraminidase from Vibrio Cholerae demonstrate that endogenous FSH is inactivated by the enzyme. Comparisons with a zero-test group showed inhibition of antrum formation.


Plasma aldosterone in newborns and children. Plasma aldosterone concentrations were determined by radioimmunoassay in 134 healthy newborns and children of both sexes from the first day to 14 years of life. The highest mean value of aldosterone level was found during the first 24 hours of life (79.5 ng/100 ml), with a mean elevation by a factor of 10 in comparison to the results of adults. An age dependent decrease towards the children in the age group 6-14 years was found. The aldosterone concentration of the age group 6-14 years did not differ significantly from the aldosterone level of adults. The question of the origin of the increased aldosterone levels found in newborns and infancy is discussed.


Serum prolactin after electroconvulsive therapy in man. When electroconvulsive therapy without anticonvulsive premedication was administered to male psychiatric patients, serum prolactin (PRL) increased regardless to the rather high initial values due to the chlorpromazine treatment. Maximal levels were found 30 min after current application, yet 60 min after electroshock the serum PRL values were declining against pre-shock levels. Infusion of 30 mg of phentolamine during 40 min prior to electroshock was without any effects on the PRL response. The obtained data indicate that even a 5-fold higher initial serum level of PRL fails to inhibit the stress-induced hormone release and that alpha adrenergic receptors scarcely play a role in this response.


Pelizaeus-Merzbacher disease: biochemical analysis of isolated myelin (electron-microscopy; protein, lipid and unsubstituted fatty acids analysis). Analysis of myelin from a leukodystrophic brain was performed (Pelizaeus-Merzbacher disease, classical type). Myelin recovery was 7% of normal, when isolated by ultracentrifugation. Electron microscopy showed a great amount of loose lamellae, with less thick sheaths and periodicity close to normal. This myelin contains fewer lipids than normal, sphingolipids and plasmalogens being reduced. FAtty acids from phospholipids are essentially normal, however enols from plasmalogens are largely reduced. Purified sphingolipids (cerebrosides, sulfatides and sphingomyelin) present a considerable diminution in very long chain fatty acids; the ratio of very long chain fatty acids (over C18) on shorter chains is 1% of the normal value for saturated fatty acids and 2% for the monounsaturated homologues. Protein analysis showed that basic protein and proteolipids were reduced, Wolfgram proteins being relatively increased.


Charcot-Marie-Tooth disease with sex-linked inheritance, linkage studies and abnormal serum alkaline phosphatase levels. A family with Charcot-Marie-Tooth disease (CMT) is described. An irregular dominant sex-linked inheritance is observed. No linkage relationship between the Xg(a) blood group locus and the CMT locus was established. Abnormal values of the serum alkaline phosphatase level were found in affected and unaffected members.


EEG sleep study in parkinsonian patients under bromocryptine treatment. Nocturnal sleep patterns were registered from 6 parkinsonian patients treated with bromocryptine, ("-Br-L-ergocryptine, C.B.-154) a dopamine-like agonist. No significant differences in their sleep patterns were found in comparison with patients treated with L-dopa. Since bromocryptine acts direct on the dopaminergic receptor sites, it is suggested that the disturbed EEG sleep patterns in parkinsonian patients cannot be explained by altered dopaminergic or serotoninergic effects whether pre- or postsynaptic.


Liquid preservation of canine granulocytes obtained by counterflow centrifugation-elutriation. Granulocytes (PMN) were isolated from 120 ml of canine whole blood by a modification of the counterflow centrifugation-elutriation technique. Isolated cell suspensions of 96% granulocytes and 4% mononuclear leukocytes with a 21:1 PMN/RBC ratio were stored at 4 degrees C in 4:4:2 medium consisting of four parts HBSS minus Ca++ and Mg++, four parts MEM, twp parts autologous plasma, and 20 microgram/ml gentamicin for 15 days. Granulocytes were stored at concentrations of approximately 4 x 10(6) PMN/ml in polypropylene centrifuge tubes. The stored granulocyte suspensions were assayed in vitro 0, 1, 4, 8, and 15 days to monitor chemotaxis, bacterial growth inhibition, O2 consumption associated with phagocytosis, and enzyme activities. Cell volume analysis was used to evaluate cellular integrity of the liquid-stored granulocytes. Canine granulocytes isolated by the modified dilution technique of counterflow centrifugation-elutriation can be preserved for up to 15 days with 77 +/- 6% granulocyte survival with maintenance of morphological and organelle integrity, as well as retention of in vitro functions of recognition, migration, phagocytosis, and killing of gram-negative bacteria.


Incapacity of hematopoietic stem cell-deprived mice to produce tumor colonies induced by Friend virus-infected cells. The role of host hematopoietic stem cells in the formation of tumor colonies in the spleen of (C57BL/6 X DBA/2) F1 mice after grafts of spleen cells from Friend virus (FVP)-infected donors has been investigated. Hematopoietic stem cell compartments of recipient mice were destroyed by Myleran treatment or gamma-ray irradiation. A single injection of Myleran reduced the pluripotent hematopoietic stem cells (CFU) and the erythropoietin responsive cells (ERC) in polycythemic mice to around 1% of that of controls. Repeated injections of erythropoietin (EPO) restored the erythropoietic precursor cell (ERC) population. Pretreatment of polycythemic hosts with Myleran totally suppressed the tumor colony forming ability of grafted Friend virus-infected spleen cells, whereas it had no effect on tumor colonies produced by inoculation of true tumoral Friend cells. After EPO injections in such Myleran-treated recipients, with a consequent appreciable ERC repopulation, splenic colonies again occurred. Similar results were obtained in hosts whose ERC populations were damaged by irradiation. These data strongly suggest that splenic colonies result from the proliferation of the host cells transformed by virus released by Friend virus-infected cells and not from the proliferation of donor tumor cells.


Granulocyte isolation by counterflow centrifugation-elutriation of canine blood obtained by continuous-flow centrifugation leukapheresis. Granulocytes have been isolated by counterflow centrifugation-elutriation (CCE) from canine leukocyte-rich blood obtained by continuous-flow centrifugation leukapheresis (CFCL). We have attempted to define both the maximal granulocyte recovery and the efficiency of granulocyte purification for the Beckman JE6 elutriation rotor when large volumes of leukocyte rich blood are utilized. The efficiency of granulocyte purification by CCE is 81% (1.31 +/- 0.1 x 10(9) PMNL) if the number of granulocytes entered into the Beckman JE-6 rotor as leukocyte rich blood is limited to 1.60 +/- 0.13 x 10(9) PMNL. Approximately 96 +/- 2% of the leukocytes in the purified fraction were of the granulocytic series with mononuclear leukocytes comprising the residual 4% of the cell population. In vitro analysis of the isolated granulocytes indicated that the cells did not lose their morphological integrity or physiological function as a result of the dual CFCL/CCE procedure relative to granulocytes isolated by CCE from freshly drawn peripheral blood.


Teratogenic effect of windowing eggs at early stages of avian development. The chick embryo is widely used as an experimental model for embryological and teratological studies. Treatment may be iether across the intact shell or through a window cut in the shell above the embryo. However, it is apparent from the present investigation that merely windowing eggs at early stages of development is highly teratogenic. Abnormal development can be attributed to the presence of an introduced air space above the early embryo. Obliteration of this artificial air space by three different methods significantly reduced the teratogenic response.


Fat-storing cells in the liver of rats kept on a low-magnesium diet. While studying the effect of low-magnesium diet on the rat liver considerable proliferation and enlargement of cells, identified as the Ito cells, has been observed. These events were accompanied by the accumulation of lipid substances in the cells. It is the authors' opinion that the lipid accumulation in the Ito cells is due to the deficiency in the lipid metabolising and transporting enzymes. Disturbances in the production of enzymatic proteins may result from the considerable and longlasting magnesium deficiency. The contribution of the fat-storing cells to the collagen biosynthesis is also discussed.


Morphological findings in spontaneously hypertensive rats in comparison with findings after experimental renal hypertension. The morphological findings in spontaneously hypertensive rats (SHR) were compared with the findings after experimental renal hypertension. In addition the effect of an arteriosclerogenic diet was investigated. Cardiac hypertrophy occurred in all types of hypertension. During the course of renal hypertension there was also hypertrophy of the tunica media of the arterial vessels of the heart, kidneys, mesenterium, and orta. Degenerative changes in the vascular walls occurred when nephrectomized rats were given a diet containing 2% cholesterol and 1% cholic acid. This produced intima lipoidosis after seven to eleven weeks. The intensity seemed to be dependent on the degree of hypertension and the duration of the experiment.


Ethanol toxic effect on the newborn rat liver.--Histochemical and electronmicroscopical investigations. The object of the study was the liver of newborn rats. Specimens were taken from the 2nd to the 8th hour after birth. Tissue material was obtained from control animals and the newborns whose mothers had been ethanol fed throughout gestation period. 40% ethanol was administered in doses of 8.0 g/kg weight, by gastric tube. In the newborn liver ethanol ingestion had led to significant accumulation of lipids, a strong acid phosphatase reaction and to a drop in succinic dehydrogenase activity. Histochemically, the intensity of alcohol dehydrogenase activity did not show any difference when the ethanol treated newborn liver was compared with controls. Ultrastructurally, the changes in the liver cells were expressed by a disappearance of the rough endoplasmic reticulum elements. Mitochondria were often swollen and distorted.


Studies on morphological changes of arteries by administration of soluble immune complexes to low calcium diet bred guinea pigs.--Special reference to the morphological pathogenesis of arteritis. Guinea pigs bred on a low calcium diet for 5 weeks, were injected soluble immune complexes once and angitis or angiolytic changes of the vascular wall in the lungs were noted. Leucocytic adhesions along the luminal surface of the vascular wall, which were carved with leucocytic reactions, could be noticed. Loosening or duplication of the basement membrane accompanied by protrusion of vascular smooth muscle cells into the subendothelial space could be demonstrated. Above results are thought to be a consequence of the conjugation of immune complexes and the negative charges of acid mucopolysaccharides.


The nephrotoxicity and renal accumulation of amikacin, tobramycin and gentamycin in rats, rabbits and guinea pigs. The nephrotoxicity and renal concentrations of amikacin, tobramycin and gentamycin were studied in rat, rabbit and guinea pig. The nephrotoxicity was estimated by histological examination and by measuring the relative weight of the kidneys (mg/100 mg body weight). These two systems had a significant correlation (r = 0.87; p less than 0.001). The concentrations of aminoglycosides were determined by the cylinder-plate method measuring free aminoglycosides. The renal toxicity of the aminoglycosides in equal doses weight by weight followed the order amikacin less than tobramycin less than gentamycin in the rat. The same order was observed in the renal accumulation of the aminoglycosides in rat, rabbit and guinea pig. The critical kidney damaging concentration of free aminoglycoside in the whole rat kidney was estimated to be 160--190 microgram/g.


Experimental ultrastructural study on stimulation of the rat gastric endocrine cells. In vitro distension of the stomach by air inflation. The effect of distension of the stomach by air (in vitro) upon the ultrastructural picture of gastric endocrine cells in rats starved for 72 hours was investigated. Distension of the stomach by air in vitro, lasting for 5, 10, 15 and 20 minutes, resulted in massive dissolution of gastrin granules that had accumulated in the course of starvation. In AL, D1, EC and ECL cells distension of the stomach by air failed to induce dissolution of granular content or release of granules by any other mechanism. In AL and D1 cells accumulation of secretory granules caused by starvation was observed, the ultrastructure of EC and ECL cells remained unchanged both under the effect of starvation and distension. Dilatation of endoplasmic reticulum profiles as well as other changes on some cell organelles observed on endocrine cells after distension of the stomach by air in vitro are due to the effect of autolysis.


The results of family therapy revisited: the nonbehavioral methods. Studies from 1971 to 1976 reporting on the outcome of the nonbehavioral family therapies are analyzed and critically reviewed. Such research has increased in both quality and quantity since 1970 and, broadly speaking, has legitimized the status of family therapy as a viable mode of helping. Particularly potent effects were noted for family therapy as an alternative to psychiatric hospitalization, with psychosomatic problems in children and adolescents, and in certain applications with parent-child and parent-adolescent relationships. However, a number of studies comparing family therapy with no formal treatment or an alternative treatment found little difference in outcome. Problems in family therapy outcome research are discussed and some future directions suggested.


Technolatry, methodolatry, and the results of family therapy. Wells and Dezen's revisited results of research on the outcomes of nonbehavioral family therapy are themselves revisited. While their conclusions are largely defensible in terms of conventional criteria for research design and for assessing change in psychotherapy, we question whether such standard criteria are sufficient for studying the outcomes of family therapy. Moreover, Wells and Dezen's preoccupation with therapeutic technology at the expense of relationship factors and of "objective" change measures at the expense of more inferential measures severely limits the clinical and conceptual meaningfulness of their review.


Ideologies, idols (and graven images?): rejoinder to Gurman and Kniskern. Three major issues raised in Gurman and Kniskern's commentary are discussed. These are (a) the suitability of established research design criteria for studying the outcome of family therapy; (b) the impact of therapist relationship factors on therapy outcome; and (c) the place of concrete or objective change measures in psychotherapy outcome research. Areas of agreement and disagreement with Gurman and Kniskern's observations are identified.


Family ties and child placement.

Family ties and child placement. The fundamental premise of this paper is the primacy of the child's experience of biological-familial continuity in establishing his sense of self and personal significance. This paper examines the effects of current child placement practices on the child's ties to his biological, foster, and adoptive families. It explores alternative practices that would take into account biological-familial continuity. Comment is invited.


Adoption: legal resolution or legal fraud? Adoption, long considered a binding social contract, is under attack because it separates the adopted from their biological family. The attitudes and practices of secrecy by the courts, agencies, and adoptive families have intensified the adoptees' feelings of being different and cut off from their roots. In a changing climate of greater openness about sensitive subjects, less tension in discussing adoption can be a significant prophylactic factor. In this sense, adopted people, collectively and individually, are making a meaningful contribution to a complex area of human relations. However, if adoption records are unsealed by legislative acts, the primary needs of the adoptive and biological parents can be undermined by the needs of the adopted adult.


Birth parents who relinquished babies for adoption revisited. The fact that adoption records may be opened by court decree to enable adoptees to have access to identifying information about their birth parents makes it incumbent upon those concerned with adoption practices to study the impact of this on adoptees, adoptive parents, birth parents, and professional practice. This paper reports on research addressed to the attitudes and feelings of birth parents years after they relinquished babies for adoption.


Keith: a case study of structural family therapy. This is an edited case presentation of marital therapy of a couple whose child was originally presented as the problem. Two primary themes are emphasized throughout the case. The first is that therapy consists of stages involving critical transitional points that need to be appropriately timed. The second is the use of a task to bring about structural realignment within the family. Excerpts were chosen because they clearly represent the process of movement in the therapy, illustrating both the stages of therapy and the utilization of an assigned task by the therapist. Commentary on the case material is interspersed throughout transcriptions.


Family interaction and communication deviance in disturbed and normal families. Confusions and conclusions: a response to Doane. The purpose of this paper is to evaluate critically the logic and conclusions of Doane's article (14), and in so doing, to raise serious questions regarding her evaluation of the family interaction literature. In particular, we suggest that Doane's paper is characterized by (a) a confused and inaccurate presentation of previous reviews; (b) a vague and contradictory set of terms around which she attempts to recast previous findings; (c) an arbitrary and unsystematic selection of findings used to support suggestions regarding reliable group differences; (d) uncritical and erroneous interpretations of findings that ostensibly support suggested differences between groups; and (e) a superficial acceptance of recent family interaction data concerned with the relationship between communication deviance/clarity and psychopathology. We hope our detailed critique will stimulate other observers of family interaction research to contribute more accurate and significant perspectives to the field than those presented in Doane's review.


Family interaction and communication deviance in disturbed and normal families. Questions of strategy: rejoinder to Jacob and Grounds. Perhaps the best way to respond to Jacob and Grounds' critique is to focus on the more important issues they raise and to clarify some of the apparent confusion. It seems there are several general classes of criticism that Jacob and Grounds have offered: arbitrariness in selection of studies, inconsistency in reporting findings, inaccuracies in the data reported, and an overly favorable conclusion with respect to the communication deviance literature. Each of these issues will be dealt with here, using as many specific illustrations and examples as space permits.


[Biological activity of 1,2-benzylisothiazoles. Relationship between the structure and the auxinlike activity of the 1,2-benzylisothiazol-3-ylacetic acid class]. The LFER model was used to study quantitatively the interrelations between chemical structure and auxinic activity of some methyl esters of 1,2-benzisothiazol-3-ylacetic acids, using as biological response the cell enlargement induced in etiolated stems of Pisum sativum, var. Alaska. Having demonstrated that activity, expressed as log 1/C, is correlated, by Eq. 8, with factionary charge (-- Qs) calculated for the sulfur atom and with the logarithm of retention time (log t), determined by reverse phase HPLC, the AA. deal with the influences of the latter parameters on the effect registered.


[Non-steroidal anti-inflammatory agents. V. Optically active 4-biphenylyloxybutyric and 4-biphenylylglycolic acids]. The synthesis and the resolution of the enantiomers of (+/-)2-(4-biphenylyl)-4-oxybutyric and (+/-)4-(4-biphenylyl)-2-oxybutyric acids and the resolution of the enantiomers of (+/-)2-(4-biphenylyl)-2-oxybutyric and (+/-)4-biphenylylglycolic acids are described. The potassium salts of each enantiomer were submitted to preliminary assays of antiphlogistic activity.


Synthesis of some 11H-indolo[3,2-c][1,8]naphthyridines. The preparation of some 11H-indolo[3,2-c] [1,8]naphthyridines (V), having oxygen functions at positions 3 or 3 and 10 or 3 and 8, as in steroidal active compounds, is described. Compounds (V) were obtained using the Fisher indole synthesis. The preparation of the parent nucleus: 11H-indolo [3,2-c] [1,8]naphthyridines (V n) is also described. Some compounds were subjected to pharmacological screening, but none was found to be significantly active.


[Heteropolycyclic systems. VII. 4-Dialkylaminoalkyl-2,3,3a,4-tetrahydro-1H-pyrrolo[2,1c] [1,2,4]benzothiadiazine-5,5-dioxides and their benzene-substituted derivatives. Cardiovascular effects in the anesthetized rat]. A series of 4-dimethylaminoethyl and 4-diethylaminoethyl derivatives of 2,3,3a,4-tetrahydro-1H-pyrrolo[2,1-c] [1,2,4] benzothiadiazine-5,5-dioxide substituted or unsubstituted in the benzene ring was prepared and subjected to preliminary investigation using the cardiovascular system of the anesthesized rat. Introduction of a basic group in the alkyl chain on N4 gives compounds with good hypotensive activity, this being most pronounced for (III, IV a and XIV) and an increase in differential pressure, marked for (V a, X, XII). These effects are sometimes accompanied by some bradycardizing activity, most marked for (V a) and (X). The results therefore confirm that derivatives of tetrahydropyrrolbenzothiazine have cardiovascular activity. Structure-activity relationships are discussed.


[Pyrazolo[3,4d][1,3]thiazine systems].

[Pyrazolo[3,4d][1,3]thiazine systems]. The synthesis of 1,2,4,6-tetrahydro-2-methyl-3-hydroxypyrazolo [3,4-d] [1,3]thiazine-4,6-dithione is described. By spectroscopic methods the anphionic structure of this and other related compounds is demonstrated. In addition, the action of alkylating reagents upon pyrazolo-[3,4-d] [1,3]thiazine-4,6-dithionic compounds is examined and described. The biological activity of the new synthesized compounds is reported.


[Chemical and pharmaceutical research on pyran derivatives. XI. Synthesis of 2-dialkylamino-4-oxo-10-methyl-4H-naphtho[2,3b]pyrans]. Reaction of N,N-dialkylethoxycarbonylacetamides with 1-methyl-2-naphthol, in the presence of phosphorus oxychloride, gave rise to the formation of 2-dialkylamino-4-oxo-10-methyl-4H-naphtho[2,3-b] pyrans through the preliminary attack of the amide-phosphorus oxychloride reactant at the phenolic hydroxyl and cyclization at position 3 of the naphthalene moiety. However when (N-alkyl,N-phenyl)ethoxycarbonylacetamides were used in the reaction an ortho position of the N-phenyl group was involved in the cyclization and 1-alkyl-2(1'-methyl-2'-naphthoxy)-4-quinolones were achieved. Pharmacological investigation showed that some naphtho[2,3-b]pyran derivatives have neurotropic activity of the sedative, anticonvulsant and antidepressant type very similar to that shown by previously studied 1-oxo-3-dialkylamino-1H-naphtho[2,1-b]pyrans (5).


[Phytotoxic activity of N,N-di-sec-butylbenzamides with alkyl substituents on the aromatic nucleus]. A new series of N,N-di,sec.butylalkylbenzamides was studied and it was found that some compounds showed particularly interesting selective phytotoxicity comparable with that shown by analogous halogen derivatives.


[Unexpected anti-inflammatory activity of rigid structure derivatives of 6-arylpyridazinone antihypertensives. I. Synthesis and activity of 4,4a-dihydro-5H-indeno[1,2c]pyridazin-3-ones]. The antihypertensive 5-methyl-6-p.cyanophenyl-4,5-dihydro-3(2H)-pyridazinone has been embodied in a rigid framework corresponding to a 4,4a-dihydro-5H-indeno[1,2-c]-3-pyridazinonic structure (II). The resulting 7-cyano derivative (IIc) was found to be devoid of antihypertensive activity. However this compound, as well as other members having structure (II), exhibited antiinflammatory properties.


[Benzotriazole derivatives active on plant growth. I. Preparation, characterization, and correlation between physicochemical properties and structure]. Following preliminary observations of auxin-like activity for several benzotriazole derivatives, a large number of new derivatives bearing on position 1 or 2 different kinds of omega-substituted chains were prepared and tested for activity on plant growth. Some physico-chemical properties which are potentially correlated with biological activity were determined for all the new benzotriazole derivatives and for those already studied in this field as growth promoting substances. Partition coefficients (between n-octanol and buffer with pH 5.6) and RM values of 2-substituted isomers were correlated, through regressional analysis, with the corresponding properties of 1-substituted isomers. Finally, the chromatographic characterictics of the sixty substances so far considered were correlated, again through regressional analysis, with the corresponding partition coefficients and with structural features and specific functional groups by which the eventual interactions with the substratum are regulated.


[Effect of benzotriazole derivatives on plant growth. II]. The action on plant growth of sixty benzotriazole derivatives carrying on position 1 or 2 several kinds of substituents, has been studied via the oat coleoptile section elongation test. At medium and low concentrations almost all compounds behave as activators, while at the highest concentrations many of them, like indolylacetic acid (IAA), are growth inhibitors. It is worth noting the lack of toxicity, even at the highest concentration used, for the 1-hydroxyalkylbenzotriazoles and, more generally speaking, the low toxicity of hydroxyalkylbenzotriazoles and of benzotriazolylalkanoic acid amides. In the range of concentration from 10(-6) to 10(-8) M some methylalkylbenzotriazoles as well as some benzotriazolylalkanoic acids and esters are more active than IAA exhibiting from 40 to 60% of the maximal activity of IAA. The activities of methylalkylbenzotriazoles and particularly that of 2-(1'-methyl)pentylbenzotriazole are outstanding; actually at the lowest concentration tested (10(-8) M and at 10(-5) M, at which concentration this compound shows the maximum activity, the biological responses correspond respectively to more than 50 and 80% of maximal activity of IAA.


Conformational properties and in vitro antibacterial activity of some diarylsulphones. The 2-pyridylphenylsulphone, di-2-pyridylsulphone and its p-nitro and p,p'-dinitro derivatives were synthesized and tested for antibacterial activity against a number of gram-positive and gram-negative bacteria. The electric dipole moment of these compounds were also measured (in benzene, 25 degrees) and analyzed in terms of molecular conformation in solution. The derivative compounds bearing a 5-nitro-2-pyridine fragment were found to be the most active in the series examined. Among these the nitro-2-pyridylphenylsulphone was found to have a broad-spectrum effect on gram-positive and gram-negative bacteria. Results of the conformational study in combination with microbiological data allowed discussion on the possible inferences concerning the stereochemical aspects of the interaction of diarylsulphones with bacterial receptor sites.


[5H-Pyrimidothiazines. I. Synthesis 6- or 7-carboxyalkyl derivatives of 5H-pyrimido[4,5b][1,4]thiazine]. The synthesis and chemical behaviour of pyrimidothiazine derivatives with a carboxyl group in position 7 [compounds (II) and (III)] or in position 6 [compounds (IV) and (V)] are described. It has been shown that these products, analogous with the isosteric benzothiazines, have the 5H-pyrimidothiazine structure. Compounds (II), (III) and (IV) show immunological activity, inhibiting passive cutaneous anaphyllaxis in the rat.


[Some aroylhydrazones of halobenzaldehydes and halogen-substituted 2,5-diaryl-1,3,4-oxadiazoles]. The preparation of a series of aroylhydrazones of halogenobenzaldehydes (I) - (XI) is described together with that of the corresponding reduction products (XII) - (XXV) and of 2,5-diaryl-1,3,4-oxadiazoles with halogen substitution (XXVI) - (XLIII). The results of pharmacological and microbiological examination of the compounds is reported.


[Non-steroidal anti-inflammatory agents. VI. Optically active 3-(4-biphenylyl)-3-oxybutyric, valerianic, caproic and isocaproic acids]. The synthesis of the (+/-) 3-(4-biphenylyl)-3-hydroxybutanoic, pentanoic, hexanoic and isohexanoic acids, and their resolution in to enantiomers are described. The latter were submitted to preliminary assay for antiinflammatory activity: interesting results were obtained above all for the isohexanoic acid derivative.


Synthesis and biological activities of some cephalosporin derivatives with carbonylamino or acetylamino [(nitro or amino) imidazole] substituents in position 7. The synthesis and the physicochemical properties of some cephalosporin derivatives with a carbonylamino- or acetylaminoimidazole moiety in position 7 are described. The effects on biological activity of the presence of some groups (methyl or ethyl, 2-nitro or 5-nitro, 2-amino) on the imidazole nucleus are examined. Selected heterocyclicthiomethyl substituents were also introduced in the 3 position of the cephalosporin nucleus. The in vitro antimicrobial activities of the compounds and their effectiveness in protecting against bacterial (S. aureus) infections in mice were evaluated. Two of the compounds (V b) and (V d) are as active as cephalexin and more active than cephalothin when administered subcutaneously, while they appear to be less effective when administered orally.


[Preparation and study of the phytotoxic activity of arylalkylamides. II. N-Phenethylamides]. A series of N-phenetylamides was prepared and tested for phytotoxic activity. Comparison with N-benzylamides showed that lengthening the alkyl chain in the aralkylamide residue reduces phytotoxicity and that, also in the case of phenetylamides, phytotoxic activity is not due to inhibition of the Hill reaction.


[Study of benzimidazole and its derivatives. III. Radioprotective structure-activity relationship of various 2-alkyl (or 2-aryl) benzimidazoles]. Benzimidazoles show important radioprotecting activity. Structure-activity relationships (radioprotective activity and toxicity) are given for benzimidazole-2 derivatives.


[Asymmetric chlorobenzoic acid N,N-dialkylamides having phytotoxic activity]. In continuation of research on phytotoxic substances with selective activity, pre- and post-emergence tests against some weed species were made using a series of 3-chlorobenzamides substituted on the amide nitrogen with a sec.butyl or a 1-methylbutyl group, the second alkyl substituent being linear, branched, saturated or unsaturated. Asymmetric 3-chlorobenzamides with a tert.butyl in place of sec.butyl or with an 1-ethylpropyl in place of 1-methylbutyl and with a second alkyl residue of different nature and with variable weight, were prepared and tested for the purposes of comparison. The results show that amides with excellent phytotoxicity and selectivity of action are obtained if a sec.butyl, 1-methylbutyl or 1-ethylpropyl group is retained and the second substituent on the amide nitrogen is suitably varied. The results give further information on the importance of the steric characteristics of the substituents on nitrogen in the phytotoxic N,N-dialkylamides.


[Phytotoxic activity of asymmetric 3,4-dichlorobenzoic acid dialkylamides]. The phytotoxicity of asymmetric N,N-dialkyl-3,4-dichlorobenzamides was studied in pre- and post-emergence tests against some weed species. The amide groups were characterized by the presence of a sec.butyl or 1-methylbutyl group and a variable second alkyl group either linear, branched, saturated or unsaturated. Isomers of the above compounds were also prepared and tested as useful comparison materials; these isomers have n.butyl or ter.butyl in place of sec.butyl or a 1-ethylpropyl in place of 1-methylbutyl or other branched alkyl groups with 5 or 6 carbon atoms, always having a methyl group in the alpha position, in place of the 1-methylbutyl and a second substituent in C2 leads to C4. The results show that compounds with phytotoxicity and selective action as regards Echinochloa and Setaria are obtained if a sec.butyl group, a 1-methylbutyl or a 1-ethylpropyl group is retained and the second substituent on the N-amide is varied appropriately. The compounds with greater phytotoxicity towards the weeds were further tested against some species of agricultural interest.


[1.3-Dipolar cycloaddition products of 8-azidotetrazol[1,5a][1,8]naphthyridines with alkynes]. A series of new 1,2,3-triazol derivatives, obtained by reaction of 8-azidotetrazol[1,5-a][1,8]naphthyridine with alkynes, is described. Some of the tested compounds showed activity against hypoxia in mice.


Synthesis of (+/-) 8,12-trans-delta5-en-cis-9-keto-13,14-en-trans-15-R,S-hydroxyprostanoic acid methyl ester. The synthesis of the title compound (X) is easily achieved by suitable transformations of the lateral chains of the 1-ethylenedioxy-2-allyl--trans-3-cyclopentane acetic acid (II).


Synthesis of three metabolites of 3-hydrazino-6-[bis-(2-hydroxyethyl)amino]pyridazine. A new rearrangement of 3-[1-methylethyliden)hydrazino] derivative. The synthesis of three metabolites (I), (II), (III) , isolated from rat urine, of the hypotensive compound 3-hydrazino-6-[bis-(2-hydroxyethyl)amino]pyridazine dihydrochloride (IV), is described. The first metabolite, 3-[bis-(2-hydroxyethyl)amino]pyridazine (I) (Scheme 1) was obtained either by reaction of 6(3)-chloro-3(6)-[bis-(2-hydroxyethyl)amino]pyridazine-N-oxide (V) with hydrazine hydrate or by catalytic hydrogenation of 3-chloro-6-[bis(2-hydrxyethyl)amino]pyridazine (VI). 3-Methyl-6-[bis-(2-hydroxyethyl)amino]-2-triazolo [4,3-b]pyridazine (II) was prepared (Scheme 2) by reaction of (IV) with acetic anhydride and subsequent hydrolysis of the resutling 6-[bis-(2-acetyloxyethyl)amino]-s-triazolo[4,3-b]pyridazine (VII). The third metabolite, 3-[bis-(2-hydroxyethyl)amino]-6-isopropyloxypyridazine (III) Scheme 3), was synthesized from 3-[(1-methylethyliden)hydrazino]-6-[bis-(2-hydroxyethyl)amino]pyridazine (VIII) which in phosphate buffer (pH 7.38) or in sodium hydrogen carbonate solution, underwent a rearrangement of new type to give (III).


[Metabolism of vincamine in the rat].

[Metabolism of vincamine in the rat]. The metabolism of vincamine hydrochloride was studied in the rat after oral administration of the drug. Vincamine is almost completely metabolized, only a small fraction of the original compound being excreted in the urine. The metabolites detected in blood, urine and tissues were purified by preparative thin layer and column chromatography in several solvent systems, and analyzed by mass spectrometry. It was found that the main urinary metabolites were vincamine conjugates (sulphates and glucuronides). Two new metabolites were detected in all the biological fluids and specimens analyzed: these compounds are more polar than vincamine and their structure was characterized by mass spectrometry, I.R. and U.V. spectroscopy and confirmed by synthesis in our laboratory.


Nitrogen mustards derived from alpha-hydroxymethyleneketones. alpha-[Bis-(2-chloroethyl)]aminomethyleneketones were prepared starting from alpha-hydroxymethyleneketones and N-(2-chloroethyl)-2-chloroethanamine. Also 3-[2-(2-chloroethylthio)ethyl]aminomethylene-2-bornanone was synthesized from 2-(2-chloroethylthio)ethanamine and 3-hydroxymethylene-2-bornanone. This compound and 3-[bis-(2-chloroethyl]aminoemthylene-2-norbornanone were active against P388 lymphocytic leukemia in mice.


[Asymmetric N,N-dialkyl-3,5-dichlorobenzamides having phytotoxic activity]. Asymmetric N,N-dialkyl-3,5-dichlorobenzamides bearing on the amide N a sec.butyl group or a 1-methylbutyl or 1-ethylpropyl and a second C3 or C4 alkyl group which may be branched or linear, saturated or unsaturated, were prepared and tested for phytotoxicity in pre- and post-emergence experiments. As reference compounds were prepared and tested 3,5-dichlorobenzamides, isomers of the first series, characterized by the presence on the amide N of a n.butyl or ter.butyl in place of the sec.butyl, of a 1-ethylpropyl or other branched alkyl in place of the 1-methylbutyl and a second alkyl group (C2 leads to C4) of varying weight and nature. The results show that all the 3,5-dichlorobenzamides studied have high phytotoxic activity and good selectivity in respect of the two graminaceous weeds tested. The amides proving most active against the weeds were tested against some species of agricultural interest. The substances proving active against the weeds and atoxic for the agrarian species were characterized by the presence on the amide N of a branched C4 or C5 group and a second C3 or C4 group.


Effect of prostaglandin A on renin secretion in the dog. The role of PGA on the release of renin in the anesthetized dog has been studied. The infusion of prostaglandin for 60 minutes at a dose unable to cause pressor modifications (2 microgram/kg/min) was shown to increase the plasma renin activity (PRA); this effect was inhibited by propranolol and strengthened by aminophylline.


[Preparation of 1-(5'-nitrofurfuryl)-5-halouracils]. Some 1-(5'-nitrofurfuryl)-5-halouracils were synthetized and tested in vitro for antimicrobial activity. N.M.R. spectra of the compounds proved 1-substitution on uracil.


[Phytotoxic activity of asymmetric N,N-dialkylamides of trisubstituted benzoic acids]. Asimmetric N,N-dialkylamides of benzoic acids variously trisubstituted on the nucleus were prepared and tested for phytotoxicity in pre- and post-emergence experiments. These amides are characterized by the presence on the amidic N of a sec.butyl or a 1-methylbutyl or a 1-ethylpropyl group and a second alkyl group of varying weight and nature. For reference the Authors prepared and studied 2,3,5-trichlorobenzamides bearing on the N atom a tert.butyl and a second linear C2 leads to C4 alkyl. The substances studied generally proved very active against the weeds tested and showed marked specificity of action towards Setaria and Echinochloa. The compounds proving most active against the weeds were also tested against plants of agricultural interest. In general these asymmetric amides proved more phytotoxic against agrarian species than the corresponding N,N-di,sec.butylamides studied previously.


Studies on the mechanism of action of mitomycin C. The in vitro formation and properties of the molecular complex between mitomycin C and native DNA were examined by means of various experimental methods; the data obtained indicate that the complex is extremely weak and that the chromophoric moiety of the antibiotic is not involved in its formation. The alkylating activity of mitomycin C was also studied using 3H-mitomycin C; while monofunctional alkylation increases almost in parallel with the concentration of the antibiotic, the difunctional alkylation, causing inter-strand cross-linkages in DNA, rapidly reaches a maximum and then remains constant even when increasing the concentration of the antibiotic and monofunctional alkylation. On the basis of these results, the currently accepted molecular model of the mitomycin--DNA interaction must be revised; a new model of this interaction is presented, which is in better agreement with the properties of mitomycin C and with the latest findings on the subject.


The binding of 8-methoxypsoralen by human serum albumin. The ability of 8-methoxypsoralen (8-MOP) to bind human serum albumin has been investigated in vitro through equilibrium dialysis and fluorescence quenching. By means of the first technique it was observed that, at concentrations presumably close to those obtainable in vivo following its administration in the photochemotherapy of psoriasis, over 80% of 8-MOP was bound to serum albumin. In human serum albumin fluorescence techniques revealed a preferential site of binding for 8-MOP with a high binding constant (Ka = 0.7 X 10(5) M-1) and precise steric requirements, since small conformational variations of the protein molecule were able to abolish its affinity for furocoumarin.


[Synthesis of 2,3-diazaphenothiazines. XIV. Dialkylaminoalkyl derivatives of 8-acetylamino-2H-pyridazino(4,5b)(1,4)benzothiazine-1(10H)-one and of the isomer 8-acetylamino-3H-pyridazino(4,5b)(1,4)benzothiazine-4(10H)-one]. A description is given of some 2 or 3 dialkylaminoalkyl derivatives of 8-acetylamino-2H-pyridazino[4,5-b][1,4]benzothiazin-1(10H)-one (IV a...d) and 8-acetylamino-3H-pyridazino[4,5-b][1,4]benzothiazin-4(10H)-one (VIII a...d). The derivatives were subjected to pharmacological examination.