Differential effects of castration on LH and FSH secretion in male and female rats. Serum levels of LH and of FSH have been measured using specific radioimmunological procedures in normal controls and in male and female rats submitted to castration 1, 2, 7, 14, 21, 28 and 35 days before.  Gonadectomy is followed by a rapid increase of serum levels of LH in males, and by a delayed response in females.  The responses of serum FSH to castration are quantitatively and qualitatively similar in the two sexes.  Both in males and in females an elevation of serum FSH levels is already present 1 day after the operation.  Serum FSH continues to rise, between post-castration days 1 and 7 with a rather rapid slope, and at later intervals with a smoother progression.


Effect of ovariectomy on oestrogen-sensitive feedback elements. Serum LH and FSH were determined by radioimmunoassay in untreated and oestrogen-treated ovariectomized rats.  The animals were sacrificed 7, 14, 21 and 28 days after ovariectomy.  Oestradiol benzoate was administered in the daily dose of 50 mug on the 2 days preceeding sacrifice.  It has been found that oestrogens exert a strong inhibitory effect on the release of LH, as evidenced by a decrease in serum titers of the hormone.  Oestradiol suppressed LH almost completely in animals with both low (7 or 14 days post-ovariectomy), and high levels of serum LH(21 or 28 days post-ovariectomy).  Oestradiol failed to totally suppress serum FSH at any post-castration time considered.  Moreover, the inhibitory effect of oestradiol on FSH release decreased as post-castration time increased.  These data suggest that the feedback mechanisms which control the secretion of LH and those which control the secretion of FSH are substantially different.


Studies on hearing loss in a community with endemic cretinism in Central Java, Indonesia. An audiometric study was carried out in a community of 642 subjects severly affected by endemic goitre and cretinism.  Hearing loss was measured in 34 out of 41 subjects diagnosed as cretins, 92 normal subjects aged 5-20 years from the same community and 54 subjects (also of 5-20 years) living in a nearby control area without endemic goitre.  The excess number of hearing defects in the endemic area seems to be entirely due to the process that leads to cretinism.  There is no reason to describe deafness and deafmutism in an area with severe endemic goitre as a separate entity.  The hearing defect showed a definite greater loss in the higher frequencies than in the lower frequencies and was found in 92% of the cretins.  Deafmutism was present in 5, a loss of more than 60 db in 8, a loss of 40-60 db in 10 cretins.  A loss of 20-30 db was found in 2.2% of normal subjects in the endemic area and 1.8% of those living in the control area.  It is concluded that audiometry is a simple and significant test to establish the presence of the neurological form of endemic cretinism, which is the most prevalent form in most endemias.  The differential diagnosis and pathogenesis of the described hearing defect are discussed.


Thyroxine metabolism in the rat: effect of varying doses of exogenous thyroxine. Experiments were performed to determine quantitatively the peripheral metabolism of exogenous thyroxine (T4) in rats brought to isotopic equilibrium with doses of -131I-T4 ranging from 1 to 20 mug/100 g body weight/day.  It was found that, although the absolute amount of T4 either deiodinated and excreted as iodide in urine or excreted as T4 in faeces increased as the dose of T4 increased, the percentage of hormone excreted by either pathway at each dose level was relatively constant.  In other words, the fractional rate of deiodination is not greatly influenced by the amount of T4 administered.  A 20-fold increase in the dose of T4 resulted in only a 4-fold increase in serum T4 concentration measured 24 h after injection, but serum T4 levels were elevated considerably more than this for several hours between injections.  Nevertheless, the highest dose of T4 was not greatly thyrotoxic.  The implications of these findings in relation to the possible association between the metabolism and action of T4 are discussed.


The immunogenic properties of highly purified insulin preparations:  The clinical importance of insulin-binding antibodies. Twenty-four diabetic patients were treated with porcine protamine-insulin (NPH-insulin) containing 7-13 mmol proinsulin per mol insulin and 27 diabetic patients were treated with porcine protamine-insuline (HP-insulin) containing 0.36 mmol proinsulin per mol insulin.  75% of the patients treated with NPH-insulin and 15% of the patients treated with HP-insulin formed detectable insulin-binding antibodies.  The difference in the antibody titre in the two groups was significant.  As a group, patients treated with HP-insulin did not have significant rise in the plasma insulin-binding capacity when compared to pre-treated values.  When comparing patients with antibodies and patients without detectable antibodies no difference in the degree of regulation could be demonstrated between the two groups.  Young patients with antibodies had a higher insulin requirement per kg per day than patients without detectable antibodies. Among patients in remission those without detectable antibodies had a longer remission period than those with antibodies.  Apart from the difference in antibody formation and hence a different distribution in the groups compared, the patients treated with NPH-insulin and HP-insulin did not differ with regard to the degree of regulation, the insulin requirement or the duration of the remission period.


16-Dehydrogenase in human placenta.

16-Dehydrogenase in human placenta. Human placenta subfractions were incubated with radioactive oestriol and 16-oxo-oestradiol.  16-oxo-oestradiol was identified from the oestriol incubation.  Oestriol and 16-epi-oestriol were characterized from the incubation while 16-epi-oestriol were characterized from the incubation with 16-oxo-oestradiol.  The 16alpha-dehydrogenase has been located in the soluble fraction (105 000 g supernatant) of the human placenta.


Respiratory activity of oocytes isolated from ovarian follicles of the rat. In order to study substrate dependent respiration oocytes were isolated from antrum follicles obtained from ovaries of prepubertal rats.  The oocytes used were either surrounded by 1 to 3 layers of granulosa cells (oocyte + corona radiata), or mechanically denuded from surrounding cells (denuded oocytes).  By use of Cartesian micro-diver technique the respiratory rate could be determined in single oocyte + corona radiata cell complexes or in samples of a few denuded oocytes.  The oocytes were incubated either in an isotonic Tris-HCL buffer or in a hypotonic phosphate buffer.  The rate of respiration was determined without addition of exogenous substrate and with addition of different substrates in varying concentrations.  Cellular respiration was linear 2-4 h both in absence and presence of substrate.  Both types of oocyte preparations were unable to utilize glucose as substrate.  Lactate stimulated slightly the respiration of the denuded oocyte while pyruvate and oxalcetate increased markedly the respiratory activity of both types of oocyte preparations.  Succinate had an influence on the oocyte + corona radiata only when the hypotonic medium was used while it was easily utilized in both media by the denuded oocyte.  In a preliminary series of experiments the in vitro influence of bovine lutienizing hormone (LH) on the cellular respiration was studied.  Both in presence and absence of corona radiata a significant decrease in oxygen uptake was registered, consistently appearing within 30 min after the addition of the hormone.  As the relationship of the phenomenon to ovum maturation was not studied, the possible biological meaning remains to be elucidated.


DNA and protein synthesis of prostatic cultures in relation to histological response under the influence of testosterone and its metabolites. Testosterone as well as dihydrotestosterone and 5alpha-androstane-3beta, 17beta-diol were capable of maintaining, in rather low concentrations (0.02, 0.02 and 1.0 mug/ml, respectively), the histological structure of the rat ventral prostate in organotypic tissue culture.  By means of histometrical analysis testosterone and dihydrotestosterone were found to be active in lower concentration (0.05 mug/ml) than adrostanediol.  In addition to the visual evaluation of the structure of the explants their total DNA and protein content was measured as well as their rate of incorporation of tritiated thymidine and leucine.  In a concentration of 1 mug/ml the response of the ventral prostate culture was almost equal to all the steroids.  Androstanediol, however, caused no stimulation in lowest concentrations (0.02-0.1 mug/ml) and was not toxic in the highest concentrations (100 mug/ml) at which testosterone and dihydrotestosterone were definately cytolytic.  When the serum from the culture medium was omitted even hormone concentrations close to the physiological level (0.003 mug/ml) could stimulate thymidine and leucine incorporation into the cultured ventral prostate.


Metabolites of lynestrenol acetate in the bile of rats after intravenous administration; a comparison with lynestrenol. Some aspects of the metabolism of lynestrenol acetate, an orally active contraceptive compound, were studied in female rats.  Lynestrenol acetate is stable in gastric and intestinal juice in vitro.  After intravenous administration of lynestrenol acetate and lynestrenol with a -14C label in the nucleus approximately 40% of the administered radioactivity was excreted in the bile within 90 min.  After administration of lynestrenol acetate labelled in the ester group, 6% of the radioactivity was found in the bile.  This means that the greater part of the lynestrenol acetate had lost its acetate group during the process of metabolism.  There was an important difference between the autoradiograms of the thin layer patterns of post-hydrolysis extracts after administration of [4-14C]lynestrenol acetate and those after administration of [1'-14C]lynestrenol acetate and [4-14C]lynestrenol: the major metabolite of [4-14C]lynestrenol acetate did not appear on the autoradiograms of [1'--14C]lynestrenol acetate and [4-14C]lynestrenol.  This indicates that lynestrenol acetate was altered in the nucleus in the presence of the acetate group.  The acetate group itself was removed, either when the alterations took place, or after it had been completed.  The results of IR, NMR and mass spectrometry analysis indicate the introduction of a 15alpha hydroxyl group.  Results of gas-liquid chromatography and thin layer chromatography indicate that a second important metabolic is 19-nor-17alpha-pregn-20-yne-3alpha, 17beta-diol.  The main conclusions are: 1. A part of the lynestrenol acetate is metabolized and excreted in the bile, the acetate group still being present.  2. Lynestrenol acetate is to some extent metabolized via another pathway than lynestrenol.  This indicates that esterification of a steroid can lead to deviation from the metabolic pathway of the free original steroid.


An improved technique for transauricular hypophysectomy and auto transplant of isolated pieces of pars intermedia tissue in the evacuated pituitary capsule of the rat. An improved technique for transauricular hypophysectomy in rats, weighing 120-300 g, is described.  This method can be used for transplantation studies, which is demonstrated by transplanting pars intermedia tissue into the evacuated pituitary capsule.  Our technique has been carried out, successfully, in various laboratories.  The success, however, depends upon the skill and care of the investigator.


Ophthalmopathy associated with primary hypothyroidism. Ten patients with primary hypothyroidism and ophthalmopathy are reported.  None of the patients had previously received any form of therapy for thyrotoxicosis.  In some patients hypothyroidism and exophthalmos presented simultaneously, but in others one clinical feature preceded the other by many years.  Elevated titres of thyroid antibodies were found in 7 of the 10 patients and LATS was detected in 1 of the 10 patients.  Increased 35S-sulphate uptake by the mouse Harderian gland technique occurred with serum from 1 of the 7 patients tested.


Glucose metabolism during and after pregnancy in normal and gestational diabetic women.  1. Influence of normal pregnancy on serum glucose and insulin concentration during basal fasting conditions and after a challenge with glucose. Glucose and insulin concentrations during basal fasting conditions and after an oral challenge with glucose have been studied during early, mid and late pregnancy and also after delivery in a group of 9 normal women.  No significant changes in the fasting serum glucose concentration was observed during pregnancy.  In contrast the fasting serum insulin gradually increased.  No changes in the mean glucose concentration curve were observed until the second half of pregnancy where the level of the curve was significantly elevated, but statistically calculated limits of normality derived from a special study of non-pregnant normal controls were not exceeded.  The serum insulin response to glucose was significantly increased at all stages of gestation and in parallel the insulin-to-glucose index calculated for the total areas below the insulin and glucose concentration curves increased significantly.  The fasting insulin-to-glucose index also increased and was found to be significantly correlated to the stage of gestation.  The shape of the glucose and insulin curves was modified in the opposite direction by pregnancy:  the peak value of glucose was delayed whereas that of insulin was advanced.  The results indicate that in pregnancy a diminished "peripheral sensitivity" to endogenous insulin apparently develops.


Inhibition of placental 3beta-hydroxy-steroid dehydrogenase by naturally occurring steroids. A potential mechanism regulating oestrogen synthesis from unconjugated precursors. We have proposed that inhibition of placental steroid 3-sulphatase by endogenous steroids may regulate oestrogen synthesis during human pregnancy. The possibility that an additional regulatory mechanism, involving the placental 3beta-hydroxy-steroid dehydrogenase (SDH), may also be operative has now been examined. Inhibitory effects of naturally occurring steroids on SDH activity were determined from the reduction in initial rate of conversion of 3H-dehydroepiandrosterone to non-digitonin precipitable products by 10 000 x g supernatant from homogenates of human term placentae. The apparent Km for dehydroepiandrosterone was 0.33 x 10(-6) M. delta4-3-Oxo products of SDH action (4-androstene-3,17-dione, app. Ki=0.60x10(-6) M; progesterone, app. Ki=1.5x10(-6) M) were the most potent inhibitors and appeared to act non-competitively. Delta5-3beta-Hydroxy alternative substrates were less inhibitory and in the case of pregnenolone (app. Ki=4.5x10(-6) M) behaved competitively. 11beta-, 16alpha-, 17alpha- or 21-hydroxylation and epimerization of 3beta- or 17beta-hydroxyl functions of inhibitors decreased their activity. It is concluded that inhibition of both sulphatase and SDH by endogenous steroids may provide complementary methods of regulating placental oestrogen synthesis in vivo. The SDH mechanism may regulate oestrogen synthesis from unconjugated precursors, either formed within the placenta or derived from the circulation. The major potential inhibitors appear to be delta4-3-ketones, acting non-competitively, and formed within the placenta. In the sulphatase mechanism alternative substrates of extraplacenta origin, acting competitively, are the major potential inhibitors controlling utilization of conjugated precursors.


Subunits of human chorionic gonadotrophin: immunological and biological studies. The alpha and beta subunits of human chorionic gonadotrophin (hCG) were prepared by incubation in 8 M urea, pH 4.5. The separation of the two subunits was obtained by DEAE-Sephadex A-25 chromatography and purification was carried out by gel filtration on Sephadex G-100. The beta subunit obtained was biologically active and was therefore further purified by affinity chromatography using as immuno-adsorbent the alpha antibodies coupled to Sepharose 4B. The beta subunit so purified showed a biological activity less than 1 IU/mg. The immunological and biological properties of the hCG subunits have been studied. It was found that the anti HCG beta serum can discriminate between hCG and hLH and that in the 125I-hCG + anti-beta serum radioimmunoassay, the cross-reactivity of pituitary hLH was lower than that of urinary hLH. Moreover, it was observed that the less purified was the urinary LH preparation, the higher was the cross-reactivity. Therefore we considered the hypothesis that during the purification of human menopausal gonadotrophin (hMG) some LH subunits or smaller immunoreactive fragments could have been discarded with the waste fractions. In order to test the validity of this hypophysis, all the protein fractions obtained during the purification of the hMG were gel-filtered on Sephadex G-100. The immunoreactivity of the effluents from the gel filtration was tested by hCG, hCG-beta, hCG-alpha and hLH radioimmunoassays. While the alpha reactive material was found in some fractions as a peak having the same Ve/Vo value as hCG-alpha, the beta reactive material presenude hMG fractions was not observed in other fractions. The cross-reactivity with the anti beta serum was very low and was found in the LH region of the gel chromatogram. Furthermore, the neutralization of the biological activity of hCG and of urinary and pituitary LH by the anti hCG beta serum was studied by incubating a fixed amount of the three hormones with increasing volumes of antiserum and measuring the LH ACTIVITY AFTER INCUBATION BY THE OADD test. It was observed that the anti hCG beta serum inhibits hCG more than urinary or pituitary LH.


Auto-transplantation of rabbit myometrium into ear-chamber and recording of its activity. A transparent ear-chamber and its installation in the rabbit is described as well as the subsequent autografting of myometrial tissue into the connective tissue formed in the chamber. Of the rabbits initially provided with ear-chambers 65% were found to be suitable for grafting while finally only 40% of these presented activity either spontaneously or induced by stretch or oxytocin. Qualitative evaluation of the mechanical activity was obtained by photo-electric recording of the transmission of light through the graft. An evaluation of the electrical activity was made possible by recording volume conducted potentials from wire electrodes in contact with the grafted tissue. A close time interrelationship between electrical potentials and change in transmission of light of the graft was found. Oxytocin produced electrical activity and blanching of the graft. It is concluded that recording of the electrical activity provides the most reliable way of expressing the activity of grafted myometrium.


Function of myometrial graft at time of parturition in the rabbit. Thirteen rabbits equipped with ear chambers containing myometrial grafts were studied during late pregnancy, spontaneous and induced parturition and early puerperium. Spontaneous electrical activity of the graft was recorded by means of F.-M. telemetry transmission in the unrestrained animals. Furthermore the reaction to intravenously injected oxytocin (threshold determination) was recorded daily. During the last 5 days before spontaneous delivery increasing spontaneous activity of the tissue was observed. During the same period the response to oxytocin increased. Concomitantly with the delivery of the litter a period of rhythmic activity was observed. This period had a sudden onset and lasted 12-55 min, while the delivery itself only lasted 6-16 min. Similar activity patterns could be evoked in the early post-partum period by 100 mU oxytocin given iv. In 3 rabbits, doses from 80-300 mU oxytocin induced complete delivery. During the 48 hours' period after delivery spontaneous activity (single spikes and series of bursts) was observed. The response to oxytocin was further increased. It is concluded that as the graft is not influenced by nervous regulation, mechanical influence or local humoral factors the spontaneous parturition and the change in excitability must therefore be due to one (or more) systemic, humoral factor(s).


The pharmacology and metabolism of testosterone undecanoate (TU), a new orally active androgen. Testosterone undecanoate (TU), either dissolved in arachis oil or as micro-crystal suspension, displays androgenic activity on oral administration to rats in the Hershberger test.  The solution in arachis oil is more active than the crystal suspension.  When [3H]TU is administered orally to rats, most of it is metabolized in the intestinal wall.  The majority of the metabolites probably lose their undecanoate group and are absorbed via the portal vein.  The non-metabolized [3H]TU and at least two metabolites are absorbed exclusively via the lymphatic system.  One of these metabolites has been identified as 5alpha-dihydrotestosterone undecanoate (5alpha-DHTU).  TU has no harmful effects on the liver of the rabbits on oral administration.


Endocrine disturbances in patients with congenital aqueductal stenosis. Congenital stenosis of the aqueduct of Sylvius is reported to be associated with sella turcia enlargement and clinical and laboratory abnormalities of the hypothalamic-pituitary-target-organ axis. It is a surgically reversible lesion. In the present report, 3 female patients with this lesion were studied with tests of basal endocrine function, as well as insulin tolerance tests, response to metyrapone and determination of circadian periodicity of plasma cortisol levels. In one patient all testing was normal and no surgery was performed. In 2 other patients the insulin tolerance test revealed either abnormal cortisol or growth hormone responses and in one patient urinary gonadotrophins were absent. All tests became normal post-operatively although in one instance not completely so until 5 years after surgery.


Adrenal potassium and sodium in experimental hyper- and hypoaldosteronism in the rat. Determination by electron probe x-ray microanalysis. Potassium and sodium contents in the various adrenal zones were determined in experimental hyper- and hypoaldosteronism in the rat by electron probe X-ray microanalysis. The analysis aimed at revealing intracellular values. There was no change in the potassium content of the zona glomerulosa, zona fasciculata and medulla neither in hyperaldosteronism induced by Na-deficiency, nor in hypoaldosteronism, elicited by Na-rich diet. The sodium content in the zona glomerulosa and zona fascicularis was increased in the Na-loaded rats, while that of the medulla was not changed. Our data are not consistent with the hypothesis that a change of adrenal intracellular potassium would act as a final stimulus in the regulation of aldosterone secretion.


Metabolism of (4-14C)progesterone and (7alpha-3H)pregnenolone by human ovaries perfused in vitro. Nine human ovaries were perfused in vitro with [4-14C]progesterone and in addition one ovary with [7-3H]pregnenolone. From the perfusate unchanged progesterone and five different metabolites were isolated: 20alpha-dihydroprogesterone, 17alpha-hydroxyprogesterone, 16alpha-hydroxyprogesterone, 5alpha-pregnane-3,20-dione and 4-androstene-3,17-dione. In the ovarian tissue saturated pregnane derivatives were the main metabolites. When [3H]pregnenolone and [14C]progesterone were perfused simultaneously a stimulation of delta4-5-isomerase and 3beta-dehydrogenase activity by HCG was demonstrated.


The binding of insulin and somatomedin A to human placental membrane. A particulate membrane fraction from human placental membrane was shown to be rich in binding sites not only for insulin but also for somatomedin A. The binding of the 125I-labelled peptide was time and temperature dependent. Degrading activity present in the membrane fraction was negligible at +4 degrees C. The Scatchard plot for insulin binding revealed two types of binding sites with an apparent high affinity constant of 3.8 times 108 M(-1) and with 5.4 times 10(-9) moles of binding sites per mg of membrane protein. The Scatchard analysis of somatomedin A revealed two classes of binding sites with an apparent high affinity constant of 2.7 times 107 M(-1) and with 1.9 times 10(-8) moles of binding sites per mg of membrane protein. In high concentrations insulin interfered with the specific binding sites for somatomedin A and vice versa. In comparison with insulin the somatomedin A preparation was one million times more potent in displacing labelled somatomedin A than in displacing labelled insulin from their respective binding sites. A radioreceptor assay utilizing particulate placental membrane and labelled somatomedin A purified on the membrane enabled the determination of somatomedin in unextracted serum. The mean values of somatomedin A in sera from patients with pituitary dwarfism and acromegaly were 0.57 and 3.2 U/ml, respectively by radioreceptor assay and 0.41 and 1.61 U/ml, respectively by bioassay. Various causes of this discrepancy between the methods are discussed.


Identification and quantitative determination of steroids in bovine corpus luteum during oestrous cycle and pregnancy. Neutral steroids in bovine corpus luteum were isolated by liquid-gel chromatography on hydrophobic Sephadex, and were analyzed by computerized gas chromatography-mass spectrometry. The presence of progesterone and 20beta-hydroxy-4-pregnen-3-one was confirmed. In addition, 3beta-hydroxy-5-pregnen-20-one, 5-pregnene-3beta,20beta-diol, 3beta-hydroxy-5alpha-pregnan-20-one and 5alpha-pregnane-3beta,20beta-diol were fully identified, and 3-hydroxy-4-pregnen-20-one, 4 pregnene-3,20-diol, 22-hydroxycholesterol and 20,22-dihydroxycholesterol were partially characterized. Steroid sulphates were not detected. Quantification of the six fully identified steroids was based on peak areas in specific fragment ion current chromatograms constructed by the computer. During the 4th-19th day of the oestrous cycle the steroid concentrations varied as follows: progesterone 6.0-36.7 mug/g wet luteal weight, 20beta-hydroxy-4-pregnen-3-one 0.8-5.5 mug/g, 3beta-hydroxy-5-pregnen-20-one 1.0-7.1mug/g, 5-pregnene-3beta,20beta-diol smaller than 0.2-0.9 mug/g, 3beta-hydroxy-5alpha-pregnan-20-one 1.7-8.6 mug/g, and 5alpha-pregnane-3beta,20beta-diol smaller than 0.2-1.2 mug/g. The concentrations of progesterone and 3beta-hydroxy-5-pregnen-20-one seemed to vary in parallel and were low during days 11-17. During this period the concentrations of 5-pregnene-3beta,20beta-diol and 5alpha-pregnane-3beta,20beta-diol were highest as was the relative contribution of all three 20beta-hydroxysteroids to the total amount of steroids. The relative amount of 3beta-hydroxy-5alpha-pregnan-20-one seemed to be highest during days 4-6. The total steroid concentration in corpora lutea taken in early pregnancy (75-105 days) was 18-47 mug/g. In the period 75-90 days, progesterone constituted only 35-42% of the total steroids, 3beta-hydroxy-5alpha-pregnan-20-one as much as 23-40% and the total 20beta-hydroxysteroids 18-30%. The total steroid concentration in corpora lutea taken in midterm and late pregnancy was 21-77 mug/g. In this period progesterone was by far the predominant steroid and constituted about 80-90% of the total steroids in corpora lutea taken between days 150 and 240. Possible correlations between luteal growth, steroid oxidoreductases and steroid concentrations are discussed.


Suprasellar disturbance in the syndrome of fertile eunuchoidism: case report. A case of fertile eunuchoidism is presented. The diagnosis was established on the basis of low androgen secretion in the presence of active spermatogenesis, increase in testosterone output after gonadotrophin stimulation, and adequate peripheral response to exogenous testosterone. Resistance to clomiphene stimulation but normal pituitary response to LH-RH was elicited indicating a suprasellar disturbance as the cause of the disorder.


Age related changes in free plasma testosterone, dihydrotestosterone and oestradiol. The age related changes on free plasma testosterone (T), dihydrotestosterone (DHT) and oestradiol (OE2) were determined in 82 healthy adult males. Forty-six subjects were between 22 and 61 years of age (group I), 36 between 67 and 93 years (group II). The percentage of free, non protein bound hormone was determined by equilibrium dialysis of undiluted plasma against isotonic phosphate buffer at 37 degrees C. Total hormone concentrations were measured by radioimmunoassay. The percentage of free T was 2.24% (median), 1.65-3.42 (95 percentiles) in group I and 1.65% (1.24-2.26) in group II. The percentage of free DHT decreased from 1.17% (0.80-2.03) in group I to 0.83% (0.52-1.55) in group II. The decrease in % free Oe2 was only very small: group I = 2.49% (2.13-2.96), ad group II = 2.31% (1.95-3.17). The fall in free T by 43.3% from 12.2 ng/100 ml (6.74-25.0) in group I to 6.90 ng/ml (3.57-10.6) in group II was twice as high as that of total T, which decreased on an average by 20.6%. Free DHT decreased by 25.8%: group I = 578 pg/100 ml (266-987), group II =429 pg/100 ml (168-723), while total DHT was not significantly different between the two groups (-1.9%). Free Oe2 was increased in old age: group I = 42.4 PG/100 ML (26.0-69.4), GROUP II = 55.7 PG/100 ML (35.8-118.9). The increase in free Oe2 by 31.4% was almost as high as that of the total Oe2 (46.9%).


The effect of oestradiol-3N-bis-(2-chloroethyl)carbamate-17beta-phosphate (estracyt) on the 5alpha-reductase in the rat ventral prostate. The ventral prostate of the rat both after in vitro incubation and in vivo experiments was found to contain appreciable 5alpha-reductase activity, whilst a very low activity was registered in the diaphragm and liver. Neither Estracyt nor LEO275 (Estracyt without the phosphate group in position 17 of the oestradiol moiety) had an inhibitory effect on the enzyme activity after in vitro incubation but equivalent amounts of oestradiol-17beta and oestradiol-17beta-phosphate significantly reduced 5alpha-reductase activity. When Estracyt was injected in vivo no influence on activity was registered in "short term" experiments while a significant inhibition was found after "long term" treatment in vivo. Possible explanations for this "long term" effect of Estracyt on 5alpha-reductase activity are discussed.


Effect of pentobarbital anaesthesia on luteinizing hormone levels in intact and ovariectomized ewes. In six intact ewes, deep anaesthesia lasting 1-2 h was induced with pentobarbital given intravenously at the onset of oestrus. In a second group of six ovariectomized ewes, light pentobarbital anaesthesia was induced following a control period of 4 h and then maintained for 2, 3, or 4 h. No drug was given to control animals (seven in the first group, three in the second). Serum levels of luteinizing hormone (LH) were determined by radioimmunoassay hourly in the first group, and at intervals of 5 or 10 min in the second. In the oestrous ewes, pentobarbital did not prevent or modify the expected LH surge. In the ovariectomized ewes, the drug, while not affecting the pulsatile release pattern of LH, reduced average LH levels to about half their control values (P smaller than 0.001) when given in a dosage maintaining 3 or 4 of light sleep; the 2-h dosage had no such effect. These differential effects of pentobarbital indicate that LH secretion in ewes is regulated by more than one mechanism. In particular, our findings are consistent with a hypothesis that there are, in ovariectomized ewes, independent central regulatory mechanisms for the amount of LH released, and for the time pattern of its release.


Radioimmunoassays of 3,5,3'-triiodo-L-thyronine with and without a prior extraction step. Two radioimmunoassays for triiodothyronine (T3) are described, one of which includes an extraction step, while the other does not. To raise antibodies, two carrier proteins and different coupling agents were used, namely haemocyanin and diazotized benzidine or human serum albumin and carbodiimide. In the case of T3 coupled to haemocyanin by diazotized benzidine, evidence of covalent binding of the hapten to the protein was obtained. In the case of T3 coupled to human serum albumin, little evidence of covalent linkage was available. Nevertheless immunization was successful in both cases. The radioimmunoassay in unextracted serum was highly reproducible and precise (intra-assay variability 5.2% inter-assay variability 8.1%). Normal values were determined which clearly indicate a fall in the serum T3 concentration with increasing age. In men the fall occurs in the fifth decade. In women the T3 starts to fall only after 70 years of age. In 31 cases of hyperthyroidism the serum T3 concentration ranged from 2.26 to 10.4 ng T3/ml. In 10 cases of hypothyroidism the values ranged from 0 to 0.8 ng T3/ml. The radioimmunoassay using an extraction procedure was less extensively used since it was found to be less reproducible (intra-assay variability 7.5%, inter-assay 12.25%). The normal values were determined with a mixed population aged 20-50. The mean +/- 2 SD was 0.9 /- .36 ng T3/ml (n = 52). In 17 cases of hypothyroidism the values ranged from 0 to 0.6 ng T3/ml and in 22 cases of hyperthyroidism from 2 to 14.4 ng T3/ml.


A comparative study of serum growth hormone and plasma cortisol levels in stimulation tests with insulin and propranolol-glucagon. Insulin and propranolol-glucagon stimulation tests were carried out on 28 children and 5 adolescents and the results of their growth hormone and plasma cortisol estimations were compared. Twenty-nine subjects with normal growth hormone reserves showed a mean maximum rise of 17.4 muU/ml of serum growth hormone in the insulin test whereas the intramuscular injection of glucagon after oral premedication with propranolol produced a rise of 38.5 muU/ml. Five subjects with normal growth hormone reserves showed a reduced hormone output in the insulin stimulation tests but normal response in the propranolol-glucagon stimulation tests. Only one subject showed a poor response in the propranolol-glucagon but normal response in the insulin stimulation test. In 30 subjects with normal adrenocortical function the mean maximum increase of plasma cortisol was 15.6 muU/ml in the insulin - and 14.9 muU/ml in the propranolol-glucagon stimulation tests, respectively. Both methods are suitable for studying the pituitary-adrenocortical interrelationships. The mechanism of the release of glucagon-induced growth hormone is not clear but the fall in blood glucose does not seem to play a major role in the process. A stress-like mechanism is equally unlikely because vegetative symptoms occurred only i a small number of subjects after intramuscular glucagon administration. It is possible that glucagon possesses a releasing-like mechanism which operates in the pituitary itself.


131I-19-iodocholesterol scintigraphy of the adrenal cortex. 131-I-19-iodocholesterol scintigraphy of the adrenal cortex has been carried out in 26 patients. In 4 patients with normal adrenocortical function the tracer was equally accumulated on the two sides. In 7 patients with untreated Cushing's syndrome, bilateral uptake was found in 4 patients with bilateral hyperplasia whereas unilateral visualization was obtained in three cases of cortisol producing adenomas. The side localization was confirmed at operation. Eight patients had been operated for Cushings syndrome prior to the scintigraphy. Remnant adrenocortical tissue with negligible or subnormal function (4 patients) could not be visualized. Normo- or hyperfunctioning remnant tissue was visualized in 3 patients. One patient had recurrent hypercorticism due to metastases from a previously removed adrenocortical carcinoma; a single pelvic accumulation was seen, whereas several metastases in the abdomen and thorax were not visualized. Four patients with aldosteronism were investigated. Three had primary aldosteronism due to an adrenocortical adenoma. In two of these, the site of the adrenal lesion was localized pre-operatively. In the third patient, equal bilateral accumulation of iodocholesterol was seen even after suppression with dexamethasone. At operation a small tumour was found. In 1 patient with indeterminate aldosteronism both glands were visualized and at a second examination the uptake was equally suppressed by dexamethasone.


New bronchodilator aerosol, salmefamol, in asthma. Twenty-five asthmatics were tested with salmefamol aerosol (200 mug q.d.s.) for a period of 3 months. The ventilatory capacity before and after adrenaline was measured weekly and symptoms were assessed daily using a scoring system over a period of 9 months. The results during the 3 months' treatment with salmefamol were compared with the preceding and succeeding 3-month-periods when patients were receiving either salbutamol aerosol or orciprenaline aerosol. Statistically significant improvements were seen in ventilatory capacity before adrenaline inhalation and in symptom scores while on salmefamol. Ventilatory capacity after adrenaline inhalation remained unchanged throughout the study: thus there was no evidence of tachyphylaxis. A significantly greater number of patients preferred the new drug. Four patients developed slight muscle tremor in the first few days of salmefamol therapy, but there were no changes in haematological or biochemical values after 3 months' therapy. Thus, salmefamol seems to have marked efficacy with low toxicity and is generally well tolerated.


Antigen-induced lymphoblast transformation in the diagnosis of cow's milk allergic diseases in infancy and early childhood. The in vitro lymphocyte stimulation of 17 milk-allergic and eight control children was carried out with beta-lactoglobulin, bovine serum albumin, bovine hair extract and phytohaemagglutinin. The blastic transformation was significant with at least one of the milk antigens in 15 of the patients, but only in one of the controls. In two clinical pictures, corresponding to milk allergy of Arthus type (Schönlein-Henoch purpura and Heiner pneumonitis), besides the high ratio of specific blastic transformation of the lymphocytes the cellular reaction to PHA was pathologically decreased. A significant correlation could be demonstrated between the degrees of blastic transformation to BSA and bovine hair extract.


Comparison of allergenicity of a perfume in animals (guinea pig maximization test) and in humans (usage test). Using the Guinea Pig Maximization Test, the sensitization rate of a perfume was shown to be 10 per cent, which means a mild allergenicity rate. A usage sensitization trial was performed with the same perfume included in a soap, on 300 persons for 2-3 months. Seventy-five of these, all house-wives, were randomly selected and tested epicutaneously, after the trial. One woman reacted allergically towards the perfume. This means a sensitization rate of 1.3 per cent, which is a maximum value, as she could, theoretically, have been sensitized before.


Morphological studies on allergen induced blast transformation of sensitized lymphocytes. The morphological and ultrastructural changes occurring in blast transformed lymphocytes, isolated from patients sensitive to grass pollen and penicillin, were studied. Certain differences in the appearance of the blast cells, as evidenced by light microscopic observation, were not found by electron microscopy. The blast cells showed foamy protoplasm regardless of the nature of the allergens employed. An increased number of either single or rosette-forming ribosomes was established. Numerous vesicles were visible in the cytoplasm. The endoplasmic sacs were more numerous. Golgi apparatus was well-developed; the mitochondria were increased and had a dense matrix.


An in vitro assay of leukocyte migration inhibitory activity from human lymphocytes stimulated with concanavalin A. Human venous blood lymphocytes, incubated for 22 h in serum-free culture medium with the plant mitogen concanavalin A (Con-A), elaborated products, which inhibited the migration of human buffy coat cells under agarose. Con-A was removed by applying the supernatants on small Sephadex G-100 columns. The leukocyte migration inhibitory activity (LMIA) was tested in a semi-quantitative modification of the indirect leukocyte migration agarose technique, which is described. Lymphokine activity, demonstrable as early as 9 h after activation of lymphocytes, was most pronounced after 22 h. Significant LMIA was demonstrated in 12 of 17 normal individuals at standard dilution of culture supernatants I/3. In 9 of the 12 experiments, assays of LMIA were carried out on stepwise diluted supernatants with detection of the greatest dilution with significant LMIA. In four experiments LMIA could only be detected after 3- to 12-fold concentrations of supernatants. Considerable individual variation was found, the amounts of LMIA varying by a factor of about 300. The reproducibility appeared to be quite high, but the factor stability of supernatants stored at -20 degrees C was surprisingly low.


Cutaneous reactions to pollen extracts in patients with hay fever. Special references to grass pollen recommended for treatment in fixed combination. Skin reactivity to 8 common grass pollens was investigated in patients with hay fever using commercial, unfractionated extracts in serial dilution technique. The highest degree of sensitivity was observed for cocksfoot, meadow fescue, meadow foxtail, rye grass, timothy and Yorkshire fog. Five of those are included in the Allpyral 5-Grass Mix. The cutaneous responses to rye grass pollen overlap with those to timothy in a high degree.


Electron microscopy of the adrenal medulla of the newborn dog under hypothermia and asphyxia. The left adrenal medullas of 20 (5 litters of 4) day-old dogs were studied with the electron microscope. The right adrenal medullas of the same animals had been examined by fluorescence microscopy and the results reported previously. In each litter one animal served as a coenothermic (37 degrees C) and one a hypothermic control (15 degrees C). The third animal (at 37 degrees C) was asphyxiated until twice its time of last gasp (about 32 min) and the fourth was cooled to 15 degrees C and asphyxiated for the same length of time as the third animal. The induction of hypothermia (to 15 degrees C) in unanesthetized puppies results in a decrease (32%) in dense-cored granules, an enlargement of mitochondria, and slight dilation of the endoplasmic reticulum. In coenothermic puppies asphyxiation to twice the time of last gasp is accompanied by the following changes: loss of dense-cored granules (52%); swelling of nuclei; clumping and marginal aggregation of nuclear chromatin material; dilation of the endoplasmic reticulum; swelling, loss of cristae and rarefaction of the matrices of mitochondria. The changes in some cells were greater than others. Many of these changes are considered to demonstrate anoxic damage. Under the conditions of the present experiments, hypothermia protected most of the adrenal medullary cells from anoxic damage.


The value of an ophthalmic tumour centre. An account is given of the clinician's experience of an ophthalmic tumour centre established in an institute of ophthalmic pathology serving the whole of Denmark (population 5 mill). The Tumour Centre performs supplementary preoperative examination of patients, and gives advice with respect to biopsy, attempted total extirpation, enucleation or observation. The activities of the tumour centre have proved extremely valuable as it contributes to establishing more general lines concerning biopsy, attempted total extirpation, observation, or enucleation, to the benefit of patients as well as research. It is a presupposition that the advisory activity is carried out as an intimate collaboration between ophthalmic pathology and clinical ophthalmology.


Malignant melanoma of the choroid as related to coexistent benign nevus. Ninety-five cases of malignant choroidal melanoma collected during a 15-year period have been studied with special emphasis on histological type, coexistence of benign nevus, and prognosis. Elements of benign nevus were found in 78% of the cases. The data tend to support the hyposis that most cases of malignant choroidal melanoma have their origin in a preexisting benign nevus, and that the tumours undergo a gradual change from a differentiated to a less differentiated type.


Bilateral massive gliosis of the retina involving the optic nerves. Report of a case. A 40-year old woman, blind in her right eye since childhood and in the left since the age of 15, presented a gliomatous tumoral mass filling up the eyes completely. Sections of the optic nerve of one eye showed the same cellular aspect noted in both intraocular tumors. The optic foramina had a normal size and there were no signs of Recklinghausen's disease. The diagnosis was bilateral reactive gliosis but the presence of a congenital anomaly has to be postulated in order to explain the involvement of the optic nerve in the process of intraocular gliosis.


Malignant melanoma of the choroid in an 11-month-old infant. A malignant melanoma of the choroid was observed in an 11-month-old fair-skinned Danish boy - one of the youngest cases ever published. There was no evidence of pre-existing melanosis of eyes or skin. The tumour showed histopathologically a typical "adult" pattern and was classified as a mixed tumour. The melanin content was marked. In some earlier reports on uveal tumours within the first year of life, pigmentation was sparse or lacking; a hypothesis has therefore previously been advanced: That melanomas are unable to produce melanin so early in life. The present case renders proof against this hypothesis.


Conjunctival goblet cells in patients with cystic fibrosis. In five patients with cystic fibrosis of the pancreas the mucous glandular system of the conjunctiva was studied, as changes, if any, in the conjunctival goblet cells might be applicable as a diagnostic test in questionable cases. A whole-mount technique was used, specially developed for studying conjunctival goblet cells. In all five cases the qualitative as well as quantitative goblet-cell findings were in accordance with a previously reported normal material. In particular, there were no signs of stagnated secretion.


Ouclar calcifications in primary hyperparathyroidism. Histochemical and ultrastructural study of a case. Comparison with ocular calcifications in idiopathic hypercalcaemia of infancy and in renal failure. In a case of primary hyperparathyroidism, ocular changes were observed as vacuoles in the basal cells of the corneal epithelium, and by staining procedures, calcium was found in the corneal and conjunctival epithelia, the corneal endothelium and in the sclera. By electron microscopy, needle-like crystals of calcium hydroxyapatite precursors were found intracellularly, also in keratocytes. The crystals in the epithelial cells were often confined to the nucleus. In a case of idiopathic hypercalcaemia of infancy, similar crystals were found intracellularly in the corneal epithelial and stromal cells, and in this case extracellular deposits, morphologically identical with extracellular deposits in conjunctival biopsies from patients with renal failure, were also seen. These extracellular aggregations were probably also composed of hydroxyapatite. The difference in morphology between intracellular and extracellular deposits is discussed in the light of the serum values of the mineral ions found in the three groups of cases.


Calcium oxalate crystals localized in the eye. Subretinal and retinal deposits, including deposits in the pigment epithelium. Calcium oxalate crystals, identified morphologically, chemically and histochemically, were found in the eyes of two patients, one with congenital buphthalmia and one with proliferative diabetic retinopathy. The deposits had not been observed clinically or by macroscopic examination during sectioning of the eyeballs. Generalized oxalosis could be ruled out as a source of the oxalates. The crystals were localized in the detached, degenerated retina, subretinally and in the retinal pigment epithelium. Previously, deposition in the last-named layer has been reported only once and only in a case of generalized oxalosis. The literature is reviewed, and suggestions concerning the formation of localized oxalate crystals are advanced. It is concluded that the pigment epithelium is probably involved in the production, perhaps through a changed metabolism.


Lymphoma and other lymphoid lesions of the orbit. Preliminary report. The orbit differs from the rest of the organism, excluding the central nervous system, as concerns lymph drainage. This may possible explain some of the peculiar features in lymphoid orbital lesions. The lymphoid tumours of the orbit are discussed on the basis of the classification most widely applied. An illustrative case is reported and it is concluded that even if local therapy may prove successful, patients in whom orbital lymphoid tumours have been diagnosed should be kept under constant observation with a view to prompt institution of treatment upon evidence of generalized disease.


Symptoms, signs and early course of optic neuritis. The study was an anlysis of the early course of optic neuritis based on the case histories of 185 patients, 57% of whom were females and 43% males. More than half of the patients suffered from multiple sclerosis. In 28% of the patients the etiology remained unknown. The most common initial symptom was acute decrease in visual acuity, but in 25% the onset was subacute or slow. Pain occured in 62% and preceded decrease in visual acuity in 16% of the cases. The initial attack was unilateral in 70% and bilateral in 30% of the patients. On admission, in 64% of the involved eyes, visual acuity was poor and in 73% the defect in the visual field involved the central field. The optic disc was normal in 46%, blurred and /or hyperaemic in 20%, oedematous in 23% and in 11% there was temporal or total pallor already on admission. The last finding was common in patients with bilateral optic neuritis with a slow onset. Six months after admission visual acuity was good or excellent in 56% and the visual field was normal in 45% of the involved eyes.


Later course and prognosis of optic neuritis. The study was a re-examination of 176 patients with optic neuritis. The follow-up period was for 38 patients, 6-12 months, for 52 patients, 1-5 years and for 86 patients, 6-24 years from the initial attack of optic neuritis. In 66% of the involved eyes visual acuity had again become good or excellent, but in 25% it was poor. The visual field was normal in only 38%, in 30% there was an absolute or relative central defect and in 31% a paracentral or peripheral defect. Recurrent attacks of optic neuritis occurred in one fourth of the patients and was a common finding in MS patients. Nineteen per cent of the eyes had suffered from more than one attack. Visual acuity was good or excellent in more than half, but the visual field was normal in only 29% of the eyes with more than one attack of optic neuritis. The frequency of bilateral involvement was high at the end of the follow-up period, 44% of patients had both eyes involved by optic neuritis. In 47 patients the initial attack was bilateral optic neuritis and 34% of these patient had permanently poor vision in both eyes. The initial attack was unilateral in 30 patients but the other eye became involved later. In 26% of all patients with bilateral involvement, visual acuity was permanently poor in both eyes. Bilateral papillitis was a common manifestation in young patients and in this age group the disease had a tendency towards good recovery.


Circulatory and respiratory effects evoked by hypertonic ventriculo-cisternal perfusion. The cerebral ventricular system of anesthetized dogs was perfused with synthetic isotonic CSF and, for 80 s intervals, with hypertonic CSF of various compositions. Hypertonic perfusion evoked centrally mediated marked increases in arterial blood pressure, heart rate, respiratory rate, and ventilation and, after some delay, an excitatory reaction resembling arousal. The responses were coordinated in time with the induced CSF hypertonicity, graded in relation to its magnitude, and reversible on return to isotonicity. The effects, which seemed to be elicited from periventricular structures in the brain stem, were more pronounced and consistent when CSF hypertonicity was produced by adding NaCl or Na-lactate than monosacharides to the isotonic CSF solution. Analysis of the cardiobascular responses indicated that they were caused by increased sympathetic vasoconstrictor and cardiac accelerance fibre activity and by inhibition of vagal discharge to the heart. The described pattern of response much resembles that evoked by physical exercise, a state which might lead to osmolar changes in the brain and CSF of a similar kind to that in the present study as a consequence of the pronounced work-induced arterial hyperosmolality. It is suggested that such an osmotic mechanism might constitute a "metabolic link" in the centrally mediated circulatory and respiratory adjustments in exercise.


Nutrition of the elderly at home. III. Intakes of minerals. Intakes of calcium, iron, potassium, and magnesium are reported for 77 men and 187 women aged 65 and over and living at home in N. Glasgow. Calcium intakes were below the national average. The importance of milk as a source of calcium and the relevance of calcium intake to bone disease in old age are discussed. Iron intakes were probably adequate, but those of potassium and magnesium may well be below optimum levels.


Stabilization of rat erythrocytes against the haemolytic effect of glass particles by tribenoside. Incubation of washed rat erythrocytes with minute glass particles results in haemolysis varying in degree according to the concentration of particles in the incubation medium. Cells pre-incubated with tribenoside1) display resistance to the destructive action of the glass particles. This protective effect is demonstrable at constant concentrations of tribenoside and varying concentrations of glass particles and conversely at constant particle concentrations and varying concentrations of tribenoside. Tribenoside thus also exerts a membrane-stabilizing action on red cells.


Penetration of azidocillin into the secretion and tissues in chronic maxillary sinusitis and tonsillitis. The penetration of azidocillin into the sinus mucosa and secretion as well as into tonsils and adenoids was studied. Altogehter 73 patients were included and they were arranged in four groups according to age, diagnosis and drug dose. The average azidocillin concentration was satisfactory and predictable in all tissues and well over the MIC for most upper respiratory tract pathogens studied. In the maxillary antrum secretion the values were much lower and in many cases under the MIC for hemophilus. This is thought to depend upon the drug's being bound to proteins in the secretions. The clinical results were good in all cases and the contribution of the surgical operation, particularly in providing drainage for the secretion, is obvious.


Effect of steroids on definitive localization of primordial germ cells in the chick embryo. It has been suggested that PGCs are attracted to developing gonads by a chemotactic-like agent secreted by the gonads and that this agent might be steroidal in nature. This study was undertaken to ascertain whether specific exogenous steroid hormones exert an influence on germ cell colonization of the gonads, by enhancing, inhibiting or otherwise interfering with it. Testosterone cypionate in cottonseed oil, crystalline testosterone propionate, estrone in aqueous suspension and crystalline estradiol-17beta were adminstered to chick embryos at 33 hours incubation. Normally developed embryos, those receiving cottonseed oil (vehicle for testosterone cypionate) and those receiving cholesterol served as controls. A decrease in the number of germ cells in the gonadal area at five days of incubation occurred in all groups receiving the androgens and estrogens. However, in only one group (that receiving testosterone cypionate) was this decrease found to be significant. The mean number of germ cells found in the cottonseed oil controls and the cholesterol controls closely paralleled that of the normally developed controls. Normal asymmetry in the distribution of the germ cells favoring the left side in the chick was not affected in any of the groups; however, the percentage distribution of the germ cells between the right and left gonads at this early stage appeared to be affected.


Rentrolental fibroplasia in a full-term infant. A 3,800-g infant, born after 40 weeks' gestation, developed retrolental fibroplasia, documented by indirect ophthalmoscopy. Elevated arterial oxygen tensions may have occurred during brief treatment for two episodes of cyanosis.


Functioning at the clinical-research interface: the clinical-research meeting. The authors describe their experience in carrying out clinical and psychobiological research in a therapeutic milieu setting. The clinical-research meeting, composed of clinical-care staff with secondary research responsibilities, researchers, and acutely psychotic patients, proved to be a useful mechanism for identifying and resolving inevitable problems at the clinical-research interface and enhanced the effectiveness of research implementation and patient care. The authors discuss three specific areas where covert issues threatened to undermine the work of the unit-the abrogation of research responsibility, the abrogation of clinical responsibility, and intergroup competition and envy.


Lymphosarcoma of the breast.

Lymphosarcoma of the breast. Four patients with lymphosarcoma of the breast are reported. Three had involvement of some nodes at the time of diagnosis. Violaceous skin discoloration was observed in two patients. A review of the literature indicates that although many of these patients present with disease initially localized to the breast, dissemination eventually occurs.


Arbovirus studies in Angola: Serological survey for antibodies to arboviruses. A serological survey was made comprising 4,590 human sera collected in 16 areas of Angola, in order to determine the distribution of arboviruses. The results obtained with the hemagglutination inhibition (HI) test have shown that the activity of the arboviruses is more prevalent in northern Angola, where a larger number of sera reacted in the HI test against group A and B arboviruses (7% and 32..6%, respectively). Several sera with HI reactions against Bunyamwera, group C, and Rift Valley fever viruses were also found.


Antigenic relationships of group A arboviruses by plaque reduction neutralization testing. Antigenic relationships of 20 group A arboviruses were assessed by the plaque reduction neutralization test, using highly specific hyperimmune mouse ascitic fluids and antisera. The existence of three complexes of group A viruses was verified. With rare exceptions, heterologous neutralization reactions were observed only among viruses in the same complex; however there were at least one or two immune reagents in each complex which were broadly cross-reactive within that complex.


Comparison of cardiovascular, hyperthermic, and toxic effects of para-methoxyamphetamine (PMA) and 3, 4-methylenedioxyamphetamine (MDA). Para-methoxyamphetamine (PMA) and 3, 4-methylenedioxyamphetamine (MDA) both had potent stimulatory effects on mean arterial blood pressure and heart rate in the cat. These effects were antagonized by bretylium, a compound which blocks release of norepinephrine. The increases in heart rate were also antagonized by propranolol and the blood pressure increases by phenoxybenzamine. PMA and MDA were nearly equipotent. In male albino mice, determination of 6 hr LD50's indicated PMA to be most toxic although 24 hr LD50's showed nearly equal toxicity. MDA had a potent hyperthermic effect in mice which was partially antagonized by bretylium and was blocked by haloperidol. The hyperthermic effect of PMA was not as pronounced and was followed by hypothermia.


[Triploidy as a cause of midtrimester gestosis(author's transl)]. The authors report a case of triploidy in a prematurely born child. The pregnancy was complicated by severe toxemia beginning during the second trimester and terminating in preeclampsia leading to cesarian section in the 33th week of gestation. The externally female child was underweight, hypotonic and asphyctic and died shortly after birth. She exhibited a complex brain malformation including excessive hydrocephaly, cebocephaly and aplasia of the piturary gland, and multiple dysmorphic signs. Autopsy revealed hypoplasia and dysplasia of the adrenal glands; no uterus, tubes and ovaries were present, but no testes were found. The placenta showed localized hydatidiform degeneration and was relatively large. The karyotype was 69,XXY, and with the aid of fluorescence markers, paternal origin of the additional haploid chromosome set was demonstrated. Typical findings in pregnancies with fetal triploidy are discussed and the possibility of prenatal diagnosis of triploidy by chromosome analysis in cultivated amniotic fluid cells is pointed out.


Influence of preoperative radiotherapy on postoperative morbidity in patients with endometrial carcinoma. In a total of 284 patients with endometrial carcinoma, stage I, the influence of preoperative intracavitary radiation on the postoperative morbidity was evaluated. In preoperatively radiated patients there was a postoperative increase in the number of wound healing disorders, urinary tract infections, febrile courses, and in the total number of patients with complications. Moreover, surgical difficulties were increased. These increases are statistically significant. Advantages and disadvantages of preoperative radiation are discussed. The influence of a febrile postoperative course on the 5-years-survival-rate is considered.


[Review of cervical smears in a gynaecologic department after a period of ten years under the consideration of colposcopic findings(author's transl)]. The study comprises 96042 cytologic cases within a period of 10 years. After screening women were admitted for biopsies. The histologic findings are reported. By 690 "right positive smears" 71 squamous cell cervical cancers stage Ib or more 87 microcarcinomata-1, 377 carcinomata-in-situ of the cervix, 100 dysplasias, 47 adenocarcinomata and 8 different malign tumors were found. 61 cases having histologic diagnosis outside are just mentioned. 629 cases which had been diagnosed in our hospital are described extensively. The first biopsy was nearly always taken by selective scraping of the ecto- and endocervix. Technical improvements of this method are explained. The efficiency of cytodiagnosis is especially pointed out by separating cases which could have been recognized or suspected by means of inspection or colposcopy only. 140 out of 282 carcinomata-in-situ and 7 invasive, mainly endocervical cancers (Ib-III), could only be diagnosed by a smear. In 17 woemn who turned out to have invasive cancer (Ib -III) a positive smear was the only reason for admission. Careful inspection of the cervix in the hospital, however, was sufficent to reveal the correct diagnosis. In our material the cervical smear could be of little help in cancer diagnosis of the upper genital tract such as adenocarcinoma of the corpus uteri, sarcoma and ovarian cancer. 39 of the 45 women with endometrial cancer cells in the specimen had bleeding anomalies, especially postmenopausal. The number of "false negative smears" mainly yields from histologic examination of 2415 uteri after hysterectomy and 4497 specimen after curettage of cervix and corpus uteri. In the first group 1 microcarcinoma-2 and 5 carcinomata-in-situ, in the second group which obviously is less representative 4 carcinomata-in-situ were found unexpectedly. It is also searched for cases which had a negative smear first and a positive or suspect smear later. This happened in 41 patients. The underlying lesion were 5 advanced cancers, 2 microcarcinomata-3 and 34 carcinomata-in-situ. Up to 30 months elapsed between the last negative cytologic finding and histologic diagnosis. 121 out of 811 suspect or positive smears were "false positive". Cytologic grouping III, IV and V inconsistently matched with the corresponding histologic results. The type and extension of squamous cell atypias are anticipated with little certainty. The great number of suspect specimen (group III) both in microcarcinoma-4 (6 in 78) and carcinoma-in-situ (33 in 344) was striking. Therefore we consider a histologic diagnosis to be necessary in this group as well as in group IV and V. The method of fractioned cervical scraping makes the decision of hospital admission easier. Low risk for the patient does not imply any loss in diagnostic security.


[Mycotic aneurysm of the ascending thoracic aorta. Clinico-pathological report of a case]. The case of a six year old girl with a congenital aortic stenosis is presented. A few months before being admitted to the Instituto Nacional de Cardiologia; her clinical manifestations suggested a bacterial endocarditis. The existance of an aneurismatic area in the ascending aorta and the later development of a crisis of angor pectoris, suggested a mycotic aneurism of the ascending aorta as a possible diagnosis; and the dissection or migration of septic material to the aneurism was found, it has a sacular form of 3.5 by 3.8 cm in the ascending aorta. There was also dissection into the left coronary artery at  the ostium level. Microscopic study showed areas of recent myocardial necrosis on the free wall of the left ventricle and in the lower part of the interventricular septum. The etiopathogeny, clinical manifesations, radiology, evolution and treatment of mycotic aneurism were considered. The need of an early clinical diagnosis and early medical and surgical treatment is emphasized.


Treatment of galactorrhea-amenorrhea syndrome with 2-Br-alpha-ergocryptin (CB 154). Clinical response and pattern of pituitary and steriod hormones before and during therapy. 2 Bromo-alpha-ergocryptine, a specific prolactin inhibitor, was administered to 9 patients suffering from galactorrhea-amenorrhea syndrome of varying aetiology. Plasma levels of FSH, LH, prolactin, total estrogens and progesterone were measured by radioimmunoassy before and after treatment initiation. Prior to treatment, plasma prolactin levels were in all cases supraphysiological. FSH and LH levels were, with the exception of one patient, in the low cyclic range. One patient had subnormal gonadotropin levels, presumably reflecting hypophyseal insufficiency following surgical removal of a pituitary adenoma. Mean plasma levels of total estrogens were in the lower normal range. Administration of CB 154 led in all cases to a reduction of plasma prolactin levels. In eight cases, galactorrhea was suppressed during the first month of treatment. Eight patients menstruated and seven ovulated as indicated by the basal body temperature (BBT) or plasma progesterone measurement. The postoperative hypophyseal tumor patient did not, with the exception of galactorrhea suppression, respond to treatment, presumably due to hypophyseal insufficiency. 2 patients conceived during the course of treatment. One patient, who developed galactorrhea-amenorrhea syndrome as a result of psychopharmacological drug administration received 7,5 mg/day CB 154. Prolactin secretion, as indicated by plasma levels, was inhibited but the inhibitation was much slower in onset than that exhibited by the other patients and this patient ovulated only after 5 months of treatment. Upon withdrawal of CB 154 therapy after 6 to 7 months, the patients redeveloped galactorrhea-amenorrhea syndrome, so that a definitive cure could not be demonstrated.


[Synthesis of virostatically acting N[adamantyl-(1)]-carbonic acid amides]. By introduction of carboxylic acid amide groupings into 1-amino-adamantant (I), remarkable virustatic effects have been obtained, particularly which N-(1-adamantyl) cinnamic acid amide (VIIa) and N-(1-adamantyl)-3-(4'-methoxyphenyl)-acrylic acid amide (VIIc). The synthesis of the N-(1-adamantyl)-carboxylic acid amides (IV, VII, X) could be reallzed by interaction of I with carboxylic acids (II, V, VIII) via the carboxylic acid chlorides (III, VI, IX), the latter not being isolated for the simplification of the process.


Chiral 1,4-benzodiazepine. VII. Cyclization rates of 2-(n-alpha-ammoniumacyl)-amino-5-chloro-benzophenones in the chiral 1,4-benzodiazepin-2-ones. Cyclisation rates of some S-alpha-amino acid derivatives (I--VII) into chiral 1,4-benzodiazepin-2-ones were determined under physiological-like conditions (pH, temperature) and plotted against pKa values of the corresponding alpha-amino acids. No correlation between k, i.e. t1/2 values, of the acidic precursors, and pharmacodynamic activity, as determined by some standard tests, were observed, however. Unambiguity of cyclisation, and its t1/2 values reveal benefit for physico-chemical properties of the investigated acyclic precursors as transport-forms of the chiral 1,4-benzodiazepin-2-ones with prolonged pharmacological activity.


Chemico-pharmacological studies on saponins of Panax ginseng C. A. Meyer. I. Chemical part. For the purpose of clarification of pharmacological actions of ginseng saponins we tried the isolation of ginseng saponins on a larger scale and succeeded in the isolation of eight saponins in a pure state and in sufficient amounts for the pharmacological assay. These eight saponins agreed all completely with the authentic samples of Shibata-Shoji and co-workers in molecular formula of saponins and sapogenins, sugars and TLC. The physical properties of saponins were shown. As other components of ginseng, there were isolated small amounts of panaxadiol, panaxatriol, daucosterol from the first running of column chromatography and mannitol, sucrose and glucose from the ethanol insoluble aqueous extract.


[Time of onset and regression of D-penicillamine activity, measured by rupture load of rat skin]. Two experiments were carried out to determine the period of time for a) the decrease of the load resulting in rupture of excised dorsal skin strips of the rat induced by daily oral treatment with D-beta, beta-dimethylcysteine (D-penicillamine, Metalcaptase) and b) the effect produced by D-penicillamine treatment to subside after discontinuation of the substance. 1. On the 5th day of treatment (320 mg/kg/day), the load resulting in rupture of excised dorsal skin strips of rats from an animal group decreased from 3480 p (initial value) to 2350 p. On the 9th day of treatment, the value for the load resulting in rupture dropped to 1790 p. Further decrease of the load resulting in rupture cannot be obtained even after a 2 weeks' treatment with 320 mg/kg D-penicillamine. 2. An animal group was treated with 320 mg/kg D-penicillamine for 7 days. At the end of this period of treatment, the load resulting in rupture of excised dorsal skin strips amounts to 1400 p. After discontinuation of the medication, the load resulting in rupture of the dorsal skin is determined at intervals of several days. 6 days after discontinuation of the test substance, a marked rise was observed and after 15 days, no difference was found between the initial value of non-treated animals and previously treated animals. Thus, it takes a few days only to induce a decrease of the load resulting in rupture by D-penicillamine and its recovery after discontinuing the medication.


[Spectrophotometric determination of methaqualone in biologic material]. A rapid and simple spectrophotometric procedure for the determination of 2-methyl-3-o-tolyl-4(3H)-quinazolinone (methaqualone) in biological material is described. After extraction of the specimen with chloroform and washing with 0.5 N sodium hydroxide and 0.5 N sulfuric acid methaqualone is extracted with 5 ml 50% sulfuric acid and read in a spectrophotometer at 234 nm. The procedure is suitable to determine serum levels of methaqualone after a therapeutic dose.


The solution of pharmacological problems with computers. Part 9: Kinetics of the inhibition of the enzyme morphine-N-demethylase by N-allylnormorphine. Mathematical analysis by a FORTRAN-program. In this paper, we present a method for statistical analysis of enzyme-kinetic data. This analysis allows determination of the character of the inhibition of an enzyme reaction, and computes the 95% confidence limits of the dissociation constants and of Vmax. This method involves the solution of the Michaelis-Menten equation, (see article for formula), using the Guass-Newton iteration.


DNA complexing antibiotics: daunomycin, adriamycin and their derivatives. 9-Acetyl-4-methoxy-7,8,9,11-tetrahydroxy-6,7,9,11-tetrahydroxy-o-(3'-amino-2',3',6'-trideoxy-alpha-L-lyxohexopyranoside) (daunomycin, daunorubicin, NSC-82 131, Daunoblasin), 14-hydroxy-daunorubicin (adriamycin, doxorubicin, NSC-123 127, Adriblastin), dihydrodaunomycin, their aglycones and other related compounds represent the group of S. peucetius derived antibiotics. The biosynthetic glycosides as well as some semisynthetic analogues or derivatives form strong complexes with double stranded DNA, which can be understood in terms of the known intercalation model. A clear relationship is found between the complex forming ability and the biological activity of the different compounds. The structure and configuration of the sugar moiety appear of primary importance for the stabilization of the complex and, therefore, for action. A class of side chain derivatives, namely adriamycin 14-0-esters, looks promising. Different tissue distribution of such derivatives may result in new useful anticancer drugs.


Conformational analysis of atropine.

Conformational analysis of atropine. The conformation of atropine was studied by nmr and dipole moment measurements. It was shown that there is a cis conformation between the C = O bond in the ester group and the tropanyl group. The nmr measurements indicated that the Ph--CH(CH2OH)--CO-- grouping assumes conformations I and II in equilibrium, while the conformational isomer III can be excluded. Consequently, the distance between the nitrogen atom and the carbonyl group is 4.5--5.0 A, and this determines the binding to the cholinergic receptor. This distance is substantially less than the 7 A presumed by other authors earlier.


Pharmacological action of FD-008, a new dopamine-beta-hydroxylase inhibitor. III. Effects on endogenous biogenic amine levels in rats. Effect of a new dopamine beta-hydroxylase inhibitor, 5-(4'-chlorobutyl)-picolinic acid (FD-008) on endogenous amine levels in various tissue of normotensive and spontaneously hypertensive rats were determined in comparison with its mother compound, fusaric acid, and disulfiram. FD-008 decreased norepinephrine (NE) levels in the brain and heart with a slight increase of dopamine (DA) level in the brain. NE lowering activity of FD-008 was stronger than that of fusaric acid. Disulfiram was less active and the activity was one-tenth of FD-008. FD-008 lowered NE level even when DA was increased by pretreatment with L-DOPA or iproniazid but did not cause a further significant reduction of NE when catecholamines were practically depleted by pretreatment with reserpine.


[Pharmacologic effects in combined use of oxyfedrine and beta-acetyldiogoxin]. In guinea pigs the toxic effect of beta-acetyldigoxin was not increased by simultaneously administered 3-(beta-hydroxy-alpha-methyl-phenethyl-amino)-3'-methoxy-propiophenone (oxy-fedrine). There was no evidence of a potentiation of the glycoside effect by the combination with oxyfedrine (ildamen-Novodigal). The threshold dose for arrhythmia was increased: typical glycoside tachyarrhythmias only occurred after higher infused doses. In anesthetized dogs with heart failure by overdosed pentobarbital, the therapeutic effects of beta-acetyldigoxin were sustained and enhanced by oxyfedrine.


[Effect of a protein-free hemoderivative on recovering capability of rat liver following 60 minutes of hemorrhagic shock]. The effect of the protein-free standardized blood extract from calves Actihaemyl on the concentration of ATP, ADP, AMP, lactate, pyruvate, beta-hydroxybutyrate, acetoacetate and glycogen of the rat liver after a 60 min hemorrhagic shock and recovery times of 30, 60 and 120 min has been investigated. Actihaemyl lowered the shock induced increased values of lactate and beta-hydroxybutyrate within the first 30 and 60 min after termination of shock at a faster rate than did the placebo. Consequently, the ratios lactate/pyruvate and beta-hydroxybutyrate/acetoacetate were found to be decreased. The remaining parameters showed no significant differences between the placebo and the Actihaemyl group. The results suggest that Actihaemyl increases the oxydative energy metabolism after a hemorrhagic shock.


[Anti-phalloidine activity of the silymarins silybine and disilybine]. Originating from silybin, a phenylchromanone from Silybum marianum (L.) Gaertn., a dimer -- disilybin -- was prepared. Disilybin exhibits a phalloidin antagonistic activity at least 10 times that of silybin with regards to a dose of the same weight.


[Phalloidine antagonists. 1. Effects of silybine derivatives on isolated perfused rat liver]. The protective effect of two silybin derivatives against phalloidin was tested in isolated rat livers, perfused with an erythrocyte free medium. Both liver swelling and potassium loss were examined for the evaluation of antagonism. Liver swelling and potassium loss were markedly decreased when silybin hemisuccinate was added to the perfusion medium before the application of phalloidin. In contrast, the protective effect of this compound was minimal after the mushroom toxin was partially absorbed by the liver. The protective efficiency of a dimer derivative (disilybin) was much greater than that of the monomer product. The binding of both derivatives in the liver cannot be very strong because the protective effect mainly could be washed out by a single change of the perfusion medium. Spectroscopical studies on isolated hepatocytes indicate that silybin inhibits the interaction of phalloidin with the cell surface.


[Toxicology of epidithio-dioxopiperazines. 1. Studies on toxicity of cyclosarcosylsarcosineepitetrasulfide in guinea pigs and rats]. The present paper deals with the toxic effect of a mycotoxin of the group of epidithio-dioxopiperazines: cyclo-sarcosyl-sarcosin-epitetrasulfide, particularly upon the liver of guinea-pigs and rats. The sporidesmines, related to the compound tested, cause chronic obstructive cholangitis characteristic of a disease in sheep endemic in New Zealand. This type of cholangitis was attempted to be reproduced in order to establish an experimental model of producing biliary cirrhosis in animals. The acute and subacute LD50 in rats and guinea-pigs were determined intraperitoneally and intragastrally. In neither mode of application nor in short-term nor in chronic test could we find any specific, dose dependent changes of bile ducts. Merely, parenteral administration of lethal doses led to unspecific subcapsular liver necroses obviously due to direct diffusion of the toxic agent. The discrepancy between the efficacy of the toxic grass in New Zealand and the substance tested in the study is discussed.


Stereospecificity in some central and circulatory effects of phenylisopropyl-adenosine (PIA). The effects of l- and d-stereoisomers of phenylisopropyl-adenosine (PIA) were tested on the central nervous and circulatory system. In mice l-PIA in doses of 0.1--0.2 mg/kg i.p. reduced motor activity and muscular coordination, prolonged barbiturate sleeping time and exerted a hypothermic and analgetic effect. In most of these tests even 10--20 times higher doses of d-PIA proved to be ineffective. In isolated guinea-pig heart preparation l-PIA increased the coronary flow, diminished the contraction amplitude and frequency in approximately 1/20 of the doses than did d-PIA. Blood pressure of rats was markedly lowered by l-PIA in doses of 6--15 mug/kg i.v. but not by the same dose of d-PIA. There seems to be stereospecificity for PIA in various tissues and animals in vivo as well as in isolated organs. The l-isomer is usually 10--20 times more active than the d-form. In addition to stereospecificity at receptor sites, differences in lipid solubility of the stereoisomers could explain these findings.


Study of the protection on afforded by nicergoline against the effects of cerebral ischemia in the cat. The cortical evoked potential of the two cerebral hemispheres of the cat is taken as the parameter of brain function; its course is studied during strangulation which determines its disappearance, and especially during strangulation release (recovery phase). Under these experimental conditions, the injection of 9% NaCl solution in each carotid does not modify the symmetry of recovery in the two cerebral hemispheres. The unilateral injection of increasing doses of 1,6-dimethyl-8beta-(5-bromonicotinoyl-oxymethyl)-10alpha-methoxyergoline tartrate (nicergoline) (20 to 400 mug) produces more rapid recovery on the treated side. This is also the case for the unilateral injection of 40 mg of sodium malonate. Observation of this effect in the hypoventilated, hypercapnic animal, with an already dilated cerebral network suggests that the mechanism of the protection afforded is not due to vasodilatation of the cerebral network produced by either of the two products. The effect of nicergoline disappears when a previous i.v. injection of sodium malonate has inhibited anoxic depolarization of the cellular membrane and inhibited the fall in cerebral ATP caused by ischemia. It would thus appear that the anti-ischemic properties of nicergoline, acting at the level of the central nervous system, are due to an effect on the cellular membrane or to an inhibiting effect on the metabolism of the brain cell. Supplementary experiments, involving the use of more specific pharmacological reagents should allow its action to be localized at the level of the membrane and/or of cellular metabolism.


Multivariate analysis in the pharmacological evaluation of a saluretic compound. The pharmacological evaluation of the diuretic activity of a compound -- performed measuring the urinary volume and determining its electrolyte content -- may be usefully integrated by a multivariate analysis, in order to: express a general evaluation for each animal in terms of positive or negative response; evaluate the false positive responses of the control group animals and the negative responses in the treated animals on the basis of the general response. An example of application relating to a well-known saluretic, 4-chloro-N-(2-furfuryl)-5-sulfamoyl-anthranilic acid (furosemide), is reported.


[Effect of terodiline on bronchial muscles and on tracheobronchial clearance in the rat]. Effects of 1-methyl-3,3-diphenyl-1N-tert.-butyl-propylamine-hydrochloride (terodilin, Bicor) on bronchial muscle and mucociliary function was studie in isolated airways preparations of rats. Terodilin blocked the bronchoconstrictor effect of acetylcholine but was ineffective against that caused by serotonin and bradykinin. The alteration of the dose-respone curves of acetylcholine by terodilin indicated that terodilin is a partial agonist of acetylcholine with a very small intrinsic activity. This finding may explain the ciliostimulatory effect of this drug.


The role of sex differences in the effect of anabolics on the liver. The effect of two anabolic steroids, norandrostenolone-phenylpropionate (Nerobolil) and norandrostenolone-decanoate (Retabolil) on the liver was studied in rats. Body weight, wet liver weight and the protein content of the liver homogenisates were found to increase under the effect of anabolic treatment, the most explicitely in females treated with Nerobolil. The function of the liver to metabolize hexobarbital, measured in vivo, is increased by a single dose of anabolic. The prolongation of treatment keeps on shortening hexobarbital anaesthesia only in females. Even 8 weeks after the end of treatment the effect is invariably lasting in females, in males it is not. Studies of the vaginal cycle cannot prove a decline of ovarial function. In females the hepatotropic effect of anabolic treatment, performed simultaneously with the chronic carbon tetrachloride lesion can be demonstrated. The liver weight and protein content are maintained on the control level. Under the effect of anabolic treatment the function to metabolize the effect of anabolic treatment the function to metabolize hexobarbital, which has been impaired by the lesion, remains near the level of the untreated animals. Our experiments support the observations of the inductive property of the steroids being parallel to their anabolic characteristics. Their catatoxic effect is pronounced in females. Nerobolil was found to be more advantageous from the point of view of both anabolic and hepatotropic effect. Our experiments do not suggest the possibility of liver damage during the administration of these two anabolics. The effect of the anabolics on the enzymatic induction may be of therapeutic value when adequate preparations are selected and sex differences as well as the character of the liver damage are taken into consideration.


Bioavailability of trivalent iron in oral iron preparations. Therapeutic efficacy and iron absorption from simple ferric compounds and high- or low-molecular weight ferric hydroxide-carbohydrate complexes. All available results from critical hemoglobin regeneration tests, postabsorption serum iron concentration studies, 59Fe erythrocyte incorporation and 59Fe whole-body retention measurements demonstrate that humans do absorb ferrous iron between 4 and 10 times (in the average about 5 times) better than ferric iron from therapeutic oral 50--250 mg iron doses. Ferrous sulfate iron is 3 to 4 times better available than the iron from ferric ammonium citrate or sulfate. Whereas 100 mg of ferrous sulfate iron/day are sufficient for an optimal oral compensation iron therapy and to produce initial hemoglobin regeneration rates of about 0.26 g/100 ml/day, 400 to 1000 mg of ferric iron/day are necessary for the same therapeutic effect because of the poor bioavailability of ferric iron. The ratio of the dose-absorption relationships for ferric and ferrous 59Fe was shown to decrease from 0.43 for a diagnostic 0.56 mg Fe dose to 0.21 for the therapeutic 50 mg Fe dose in subjects with normal iron stores. Absorption ratios of 0.65 for the 0.56 mg Fe dose and 0.26 for the 50 mg Fe dose were measured in subjects with depleted iron stores. At all dose levels the superior bioavailability of ferrous iron was demonstrable. A high-molecular weight ferric hydroxide-carbohydrate complex (MW similar to 30 000) was palatable but so poorly absorbed that is was practically without effect on hemoglobin regeneration even at a daily 300 mg Fe dose. Following several warnings such a useless commerecial oral iron preparation was finally withdrawn from the market. The iron from any high-molecular weight carbohydrate complex of ferric hydroxide has to be suspected to be poorly absorbed and therefore therapeutical useless, unless the opposite has been demonstrated with a reliable bioassay (59Fe absorption whole-body retention and hemoglobin regeneration test). A low-molecular weight so-called ferric hydroxide-fructose complex was shown to contain iron of more or less the same poor bioavailability as contained in ferric chloride since the iron from ferrous sulfate was about 5 times better absorable. The good absorption of ferrous sulfate iron was not further augmented by even very large oral doses of fructose since this carbohydrate did not improve the ferrous iron absorption at a fructose: Fe molar ratio of 106:1. Trivalent iron in simple compounds like ferric ammonium citrate or in low- and high-molecular weight carbohydrate complexes of ferric hydroxide is so poorly available for intestinal iron absorption in man that it cannot be used for a fast and reliable oral iron therapy with reasonably low doses as it can be easily practised with quick-lease preparations of ferrous sulfate at a 100 mg Fe2


[Effect of xylitol on increased intracranial pressure]. Increased intracranial pressure could be lowered by means of 250 ml 40% xylite (1,2,3,4,5-pentanpentol) in 17 patients, the average dosage amounted to 1.36 mg/kg, the lowering of the pressure to 54.4 plus or minus 18.4%. The effect lasted 109.5 plus or minus 35.4 min; the maximum occurring after 50.4 plus or minus 14.8 min. With the exception of 3 patients who complained of slight gastrointestinal disturbances, no side reactions were observed. Paralleling the lowering of CSF-pressure increased diuresis occurred amounting to 400% (including the infusion volume). The maxima of diuresis and lowering of CSF-pressure concur. Simultaneously we can see a highly significant lowering of urine potassium and a slightly significant lowering of sodium within the first 2 h, which after 3 h is clearly receding. There was no significant change in the values of serum electrolytes, bilirubin and transaminases (SGOT and SGPT); the residual urea was just a little lowered. The lowering of CSF-pressure after xylite surpasses the effect of the same dosage of sorbit and fructose.


[Effect of naftidrofuryl on creatinine serum concentration and on PAH/inulin clearance in peripheral arterial occlusive diseases]. The effect of 200 mg 3-(1-naphthyl)-2-tetrahydrofurfurylpropionic acid beta-diethylaminoethylester (naftidrofuryl, Dusodril) administered as a single i.v. infusion, on serum creatinine was investigated in 10 patients with occlusive disease of the peripheral arteries (average age 58.6 years). In a second group of similar patients (average age 60.5 years) determinations of PAH/inulin clearance and serum creatinine were performed prior to and immediately after treatment with 400 mg naftidrofuryl per day over a period of two weeks. Creatinine concentration did not change following acute or chronic administration of naftidrofuryl; glomerular filtration rate and renal plasma flow were not affected by prolonged treatment (2 alpha less than 0.05). An obligatorily nephrotoxic effect of naftidrofuryl can therefore be ruled out.


[New method for the exact determination of changes in the size of the leg for the quantification of therapeutic success in various angiopathies]. For a quantitative evaluation of foot or leg volume and its alterations, a new type of instrument is presented. With this apparatus exact results will be achieved in a simple and quick method, which has proved true in several previous investigations. The apparatus is also suitable for the evaluation of therapeutic results concerning swelling of the extremities. This statement is based on measurements of the foot volume of five healthy male volunteers to whom in a double-blind test were administered wto different doses of the active substance ethyl-3,5,6-tri-O-benzyl-D-glucofuranoside (tribenoside, Glyvenol); for comparison also a placebo was used. The dose-response relationship in the foot volume reducing activity of the drug was measured quantitatively. Statistical analysis of the values measured revealed a significant difference at the 1% confidence level between the effects of Glyvenol and placebo on foot volume changes. Further investigations in patients are in preparations.


[Stability of amphetaminil. 1. In-vitro studies]. The decomposition of alpha-phenyl-alpha-N-(beta-phenyl-isopropyl)-aminoacetonitrile (amphetaminil, AN 1¿) in different solvent mixtures seems to depend on their polarity. The rate of hydrolysis of amphetaminil is much higher when the compound is dissolved in a methanol/0.1 N HCl (75/25; v/v) or in methanol containing traces of concentrated HCl (36%) (97.5/2.5; v/v). Using radioactively labelled amphetaminil we were able to demonstrate the decomposition of the compound on TLC plates coated with silica gel--independent from the solvent applied. When plates with cellulose as stationary phase were used for chromatography, amphetaminil proved to be stable in cyclohexan/ethylacetate but underwent decomposition in more polar solvents, for example, methanol and chloroform, respectively.


[Bacterial growth in the perfusion medium of the isolated rat brain and its inhibition by antibiotics]. In the perfusion medium of the isolated rat brain the bacterial content was examined. The perfusion medium used contained 30% bovine erythrocytes, 2 g% bovine serum albumin and about 13 mM glucose in Krebs-Henseleit-solution. Before the perfusion of the isolated rat brain was started the medium recirculated up to two hours in the perfusion apparatus. After this, the perfusion of the isolated rat brain was carried out for four hours. At various times and at different points of the circulating system samples of the perfusion medium were withdrawn and tested for their bacterial content. Using the plate count procedure, at the end of the perfusion 10(5) to 10(6) bacteria per ml medium were found. This order of magnitude of the bacterial content suggested that it is not necessary to add antibacterial substances to the medium when perfusions lasted up to 4 hours. Nevertheless, from each sample 10 bacteria species were isolated and their sensitivity against 15 different antibacterial substances was tested. The substances were ampicillin, carbenicillin, cephaloridine, chloramphenicol, cloxacillin, erythromycin, gentamycin, nalidixic acid, neomycin, nitrofurantoin, penicillin, polymyxin B, sulfadiazine, tetracycline and trimethoprim + sulfamethoxazole. Tetracycline was most active in inhibiting bacterial growth. 1 mg and 0.5 mg tetracycline added to 100 ml perfusion medium were able to prevent an increase of the bacterial count during the perfusion experiments.


[Chromatographic studies of four water-soluble rutosides]. The water-soluble rutosides O-(beta-hydroxyethyl)-rutosides (HR), trihydroxyethylrutoside (tri-HR) and tetrahydroxyethylrutoside (tetra-HR), with and without NaCl, were separated by means of both thin-layer chromatographic and paper-chromatographic methods; separation of mono-HR was carried out using only the product with NaCl. The results of these studies show: 1. According to the procedure employed, the chromatograms of the tested rutosides showed several spots up to a maximum of 12 in each product. 2. Under certain methodical conditions the chromatograms of tri-HR and tetra-HR with and without NaCl differed; in the chromatograms of the tetra products either 12 or 9 components were detected. 7 spots were found in the chromatogram of tri-I, and 6 spots were observed in that of tri-II. The tetra-HR with NaCl was found to contain a substance which reacted with the diazonium salt fast blue salt B adopting a red coloring. This substance might be responsible for the biological effects, such as the influence exerted on yeast cell respiration. 3. At present, difficulties are still encountered in separating the substance. A comparison of its chromatograms with those of rutin, quercetin, and the bivalent phenols hydroquinone, catechol, and resorcin suggests the substance to be a mono-isomer or a di-isomer of rutin with a resorcin-like structure in the A ring. 4. Preliminary studies on yeast cell respiration using tetra-HR seem to confirm the biological effectiveness of the substance.


[Gas chromatographic analysis of benzodiazepines. 2. Diazepam and its metabolites]. A gas-chromatographic method is reported which completely resolves diazepam and its major metabolites and thus enables the specific quantitation of these compounds after extraction from serum and urine. The sensitivity limits are about 3 ng/ml if 4 ml serum or urine are extracted.


[Polarographic determination of some phenylsubstituted anticonvulsants]. A method is described to determine the anticonvulsive drugs phenobarbital, phenytoin, mephenytoin, methylphenobarbital and primidone. A serum extract is nitrated, separated by thin-layer chromatography, and the nitro compounds are determined by polarography. The rates of recovery are: phenobarbital 81%, phenytoin 79%, mephenytoin 80%, methylphenobarbital 86% and primidone 84%. The sensitivity limits range from 3.5 mug/ml plasma for phenytoin to 5.5 mug/ml for mephenytoin and methylphenobarbital. This method might be useful not only for determination of therapeutic blood levels but also for toxicology because it saves time and the expenditure of work is small. The mean deviation for mononitrated anticonvulsive drugs is +/- 2.8% (for 50 mug/ml) and +/- 11.6% (for 10 mug/ml). For phenytoin being dinitrated, the values are +/- 0.6% and +/- 7.5% for 50 mug and 5 mug, respectively). We determined blood levels of out-patients containing phenobarbital and phenytoin besides other substances. Having once established calibrating curvers internal or external standards are not required.


[Radioimmunoassay of n-butylbiguanide in human serum]. A radioimmuno assay for butylbiguanide in human serum is described. The detection limit is 1.5 ng per assay tube, the intra assay variance 3 percent, and the inter assay variability 12 percent. Antibodies to the biguanide group were obtained by immunizing rabbits with p-carboxy-phenethyl-biguanide coupled to bovine serum albumin. The affinity of the antibodies for tyrosine, urea, guanidine, butylbiguanide, phenethyl-biguanide, and p-hydroxy-phenethyl-biguanide is shown by competitive binding studies. The recovery rate is 100 percent.


Antiaccelerator and decelerator activity of eleven phenothiazine derivatives. Eleven N-substituted phenothiazine derivatives were studied for their antiaccelerator and decelerator activities on the dog heart. The antiaccelerator activity was determined in heart-lung preparation in dogs. Taking into consideration the percentage inhibition of cardio-acceleration produced by continuous infusion of 3 mug/min of adrenaline, all the drugs except chlorpromazine and 10-butyl-phenothiazine in two experiments caused more than 50 percent inhibition. 10-Propylphenothiazine induced maximum inhibitory effect, i.e. 90.1 percent in a total dose of 20 mg. Two types of effects were observed upon impulse generation in the sinus node. Acceleration was produced by chlorpromazine, 10-cyclopentyl-phenothiazine, prothipendyl, and thiazinamium, which was very short-lived, whereas other drugs produced deceleration. The correlation between antiaccelerator and decelerator properties is rather a close one.