bluuebunny/crossref_metadata_2025 huggingface dataset:
CONTRACEPTIVE TREATMENT WITH LOW DOSES OF GESTAGENS
Authors: Larsson-Cohn Ulf
Published: October 2009

I ABSTRACT

Chlormadinone acetate (CMA) in a dose of 0.5 mg and Norethindrone (NET) in doses of 0.3 mg, 0.4 mg and 0.5 mg were administred daily to different groups of women. The groups consisted of between 104 and 125 individuals and the observation time varied between 1125 and 1850 months.

The pregnancy rate varied between 0 and 3.2. There was a high frequency of irregular bleeding and amenorrhoea, causing 14.5% of the women to withdraw from trial. The cycle control seemed to be better with CMA 0.5 mg and NET 0.3 mg than with the two other doses of NET. The frequency of general side-effects was relatively low. Endometrial biopsies suggested, when cycles lasting less than 15 days were excluded, that between 70% and 80% of the vaginal flows followed periods of luteal activity. Judged by pregnanediol and progesterone determinations, the corresponding figures were 50% to 60%. This discrepancy is supposed to be due to reduced progesterone production of the corpus luteum, inducing irregular secretory changes of the endometrium. This is also supported by the fact that some women showed low plasma progesterone concentration simultaneously with irregular secretory endometrium.

As the treatment was well tolerated by a majority of the participants and as it seems to have only minor metabolic effects, it is concluded that the method is a valuable complement to the other contraceptive measures available.
STATISTICAL EVALUATION OF SYMPTOMS AND CLINICAL SIGNS IN THE DIAGNOSIS OF HYPERTHYROIDISM
Authors: Lamberg B.-A. , Heinonen O. P. , Aro A. , Viherkoski M. , Knekt P.
Published: October 2009

The occurrence of 15 symptoms and 11 clinical signs was studied in 101 hyperthyroid patients and 106 euthyroid subjects. The hyperthyroid group comprised 50 patients with toxic diffuse goitre and 51 with toxic nodular goitre. The control group included 31 subjects with non-toxic goitre. The symptoms and signs were registered in a standardised manner. The difference in the frequency of each symptom and sign between the hyperthyroid and the control group was analysed statistically.

The most discriminatory were the symptoms and signs related to excessive production of heat and to stimulation of the nervous system. The results were mostly in agreement with those of the few previously existing reports. Lid signs, however, were in both groups less frequent than in a British material. The need for standardisation in registration of symptoms and signs is emphasized.
RELIABILITY CRITERIA FOR SATURATION ANALYSIS OF STEROIDS BY COMPETITIVE PROTEIN BINDING
Authors: Reeves Billy D. , Calhoun David W.
Published: October 2009

This communication is an attempt to delineate and define reliability criteria for saturation analysis of steroids by competitive protein binding assay. The discussion of these criteria evolved from three major considerations of assay method that help to place the ultimate criterion of accuracy in proper perspective. These major considerations are: 1) the measurement system, 2) the assay design and 3) the calculations and statistical control. Such an approach permits an evaluation, both relative and absolute, for a single method or for multiple methods.
COMPUTER ANALYSIS OF RADIOLIGAND ASSAY AND RADIOIMMUNOASSAY DATA
Authors: Rodbard D. , Lewald J. E.
Published: October 2009

A computer program written in extended BASIC (BIICAC) is described for analysis of radioimmunoassay and competitive protein binding assay data. This method uses logit and log transformations to obtain a linear dose responsive curve, followed by unweighted and iterated weighted least squares regression analysis (similar to probit analysis). The standard curve is plotted by computer, and potency estimates and confidence limits are obtained for unknowns. This program has been used successfully in a fully automated data processing system using a time-sharing computer service.1
A SIMPLIFIED PROCEDURE FOR THE ASSAY OF PROGESTERONE
Authors: Johansson Elof D. B.
Published: October 2009

During one year 8000 determinations of plasma progesterone have been made, using a simple petroleum ether extraction of the plasma followed by competitive protein binding analysis. The selection of the petroleum ether is crucial for the specificity, which is acceptable for the determination of progesterone during the luteal phase of the menstrual cycle and during pregnancy. The limit of sensitivity is 0.1 ng. Only 0.25 ml of plasma is needed for the determination during the luteal phase and 0.05 to 0.10 ml during pregnancy. One technician can assay 20 samples in one day with good accuracy and a precision of 7.9 per cent in the most favourable range of measurements. In research projects involving drugs the influence of these drugs on the competitive protein binding system has to be tested. Some of the samples should be further purified by thin layer chromatography as a constant check on the specificity for progesterone.
MEASUREMENT OF TESTOSTERONE BINDING SITES
Authors: Pearlman William H.
Published: October 2009

A Sephadex assay technique, in use in our laboratory for the study of steroid-protein interactions, allows expression of the molar concentration of a testosterone-binding component of human serum of high binding affinity in terms of the molar concentration of testosterone-binding sites: examples with model calculations are provided. The presence in the assay system of competing protein of low binding affinity (i. e. albumin) and of competing steroid (i. e. endogenous, radioinert testosterone) is taken into account when estimating the testosterone-binding component. With the aid of some approximations, the distribution of bound testosterone between the testosterone-binding component and albumin is readily calculated. From this estimate and the total testosterone concentration, the serum concentration of unbound testosterone (Su) is calculated.

The concentration of the testosterone-binding component is thus estimated to be of the order of 8 × 10−7M and 1 × 10−7M in late pregnancy serum and adult male serum, respectively, assuming that there is only one binding site for testosterone in the protein molecule. The testosteronebinding component appears to play a role at least equal to that of albumin in binding endogenous testosterone in adult male serum; the testosterone-binding component is clearly dominant in this role in late pregnancy serum. The values for Su at 25° are calculated to be of the order of 3 × 10−10M and 1 × 10−11M in adult male serum and late pregnancy serum, respectively, corresponding to 1.7% and 0.27% of the total testosterone concentration, respectively. It is suggested that the physiologically effective level of circulating androgen may correlate better with the serum concentration of unbound testosterone (Su) than with that of total testosterone.
RADIO-LIGAND PROCEDURES FOR ESTROGEN ASSAY IN NORMAL AND PREGNANCY PLASMA
Authors: Korenman Stanley G. , Tulchinsky Dan , Eaton L. William
Published: October 2009

Radio-ligand assay techniques suitable for measurement of unconjugated estrone and 17β-estradiol concentrations in plasma from normal and pregnant individuals are described. The specific estrogen binding protein of rabbit uterus is employed as specific reagent and activated charcoal is used as non-specific adsorbant to separate bound from free radioactive estrogen. Steroids of interest are separated by column chromatography on celite. By these techniques, large numbers of samples can be assayed with speed and precision for both estrone and 17β-estradiol.
HYPOPHYSE UND HORMONELLES CALCIUM-REGULATIONSSYSTEM
Authors: Salzer Georg M.
Published: October 2009

A survey of the literature of the last fifty years relating to our topic indicates that the sentence from Copp (1969) »there is no evidence that the anterior pituitary or the central nervous system has any direct effect on the secretion of parathormone or calcitonin« is not based upon concrete experimental data. This is particularly evident from a morphological aspect, since considerable variations in the parathyroids have been repeately observed as the result of both hypophysectomy and the application of hypophyseal extracts. Experimental evidence has contributed little material relevant to a possible hypophyseal control of the parathyroids with the exception of the observation that after hypophysectomy the plasma calcium level remains unchanged.

Contrary to this, up to now no positive morphological changes in the C-cells of the thyroid have been determined, although experimental evidence suggests a certain hypophyseal control upon the secretion of calcitonin. All the results of our own experiments in parathyroid morphology suggest significant alterations in the glands subsequent to hypophysectomy. A histological survey shows a considerable reduction in the size of the entire organ and an increase in the connective tissue of the gland. Karyometric tests reveal a reduction of the small diameter of the nuclei of 13.5%, a quantity which corresponds to that found in the nuclei of the adrenal cortex and the follicular cells of the thyroid after the same operation. As a result of hypophysectomy it is possible under the electron microscope to distinguish alterations in the secretory structures of the parenchyma cells of the parathyroid that may indicate restricted parathormone synthesis.

In order to exclude the possibility that these changes may have been conditioned by a cessation of adrenal activity, the parathyroid morphology was analyzed also after adrenalectomy. The results obtained in this way differ fundamentally from those produced by hypophysectomy.

Morphometrical investigation of the C-cell population of the thyroid after hypophysectomy show a significant reduction in the size of the nuclei of 13.6 % and, under the electron microscope, a reduction of the cytoplasm as well as an increase in the number of calcitonin granula.

Two further test series have shown that in hypophysectomized but otherwise untreated animals the plasma calcium level is within the normal range. On the other hand the normal ability to counteract hyper- or hypocalcaemia was found to be lacking to a great extent.

All the results obtained suggest the existence of some hypophyseal influence upon the hormonal calcium regulation system.

A hypothesis is framed to the effect that the feedback: plasma calcium – parathormone and calcitonin serves, above all, the purpose of compensating an alimentary over- and under-supply of calcium, whereas the hypophyseal regulatory cycle serves to meet the endogenic calcium requirements of the organism under special circumstances, as for instance growth or lactation.
TECHNIQUES FOR THE ISOLATION OF CELLS OF THE ADRENAL CORTEX, THE ANTERIOR PITUITARY AND THE CORPUS LUTEUM: MORPHOLOGICAL AND FUNCTIONAL EVALUATION OF THE ISOLATED CELLS
Authors: Sayers George , Portanova Ronald , Beall Robert J. , Seelig Steven , Malamed Sasha
Published: October 2009

A trypsin technique for the isolation of cells of the rat adrenal, the rat anterior pituitary and the bovine corpus luteum is described. The isolated cells have been characterized in morphological and functional terms.
AUTORADIOGRAPHIC TECHNIQUES FOR THE LOCALIZATION OF HORMONES AND DRUGS AT THE CELLULAR AND SUBCELLULAR LEVEL
Authors: Stumpf Walter E.
Published: October 2009

The paper describes four autoradiographic techniques which can be recommended, not without restrictions, for the study of the cellular and subcellular hormone or drug distribution in tissues. In all of the techniques desiccated slides are used which are precoated with photographic emulsion. The techniques are (I) Dry-mounting of freeze-dried sections on emulsion precoated slides; (II) Thaw-mounting of frozen sections on emulsion precoated slides; (III) Smear-mounting on emulsion precoated slides; and (IV) Touch-mounting on emulsion precoated slides. The techniques are designed to avoid or minimize translocation of the labelled molecules during preparation and during the application to photographic emulsion. Cited examples of application of these techniques demonstrate their utility in hormone research.
EVALUATION OF TISSUE STEROID BINDING IN VITRO
Authors: Wira C. R. , Rochefort H. , Baulieu E. E.
Published: October 2009

The definition of a RECEPTOR* in terms of a receptive site, an executive site and a coupling mechanism, is followed by a general consideration of four binding criteria, which include hormone specificity, tissue specificity, high affinity and saturation, essential for distinguishing between specific and nonspecific binding. Experimental approaches are proposed for choosing an experimental system (either organized or soluble) and detecting the presence of protein binding sites.

Techniques are then presented for evaluating the specific protein binding sites (receptors) in terms of the four criteria. This is followed by a brief consideration of how receptors may be located in cells and characterized when extracted.

Finally various examples of oestrogen, androgen, progestagen, glucocorticoid and mineralocorticoid binding to their respective target tissues are presented, to illustrate how researchers have identified specific corticoid and mineralocorticoid binding in their respective target tissue receptors.
ADRENALECTOMY IN CUSHING'S DISEASE
Authors: Ernest Ingrid , Ekman Hans
Published: October 2009

Forty-four patients with Cushing's disease underwent bilateral adrenalectomy and 39 survivors were followed for, on an average, eight years and a half. There were three postoperative deaths and two other patients died within one year after operation. Thirty patients are now working but six remain invalidized. In four subjects cortisol secretion persisted in spite of "total" adrenalectomy but a clinical remission ultimately occurred. Five patients, who presented with an atypical clinical picture, failed to show any obvious changes in the distribution of fat or in the complexion. Body weight did not decrease after surgery in several patients and in many others body weight increased again after an initial loss of weight. Menstruations reappeared in all premenopausal women except two. Manifest diabetes, which was found in seven subjects, disappeared and in several other cases glucose tolerance improved.

The blood pressure, which was high in most patients, normalized or decreased in all hypertensive patients but the blood pressure still remains elevated in 22 subjects. ECG-abnormalities disappeared in 11 of 17 subjects, who presented with ST-T changes and/or high amplitude R waves. The size of the heart decreased in most patients with cardiomegaly.

A remarkable improvement in the radiological appearance of the vertebrae occurred in a 16-year-old boy but in the other patients there were no such changes. Mental disturbances, which were a main cause of preoperative disability, disappeared in most cases. Two patients, however, continue to be invalidized from mental disease, which developed during the course of the hypercorticism.

One patient presented with a grossly evident pituitary tumour and in four other subjects a large pituitary tumour developed after adrenalectomy together with hyperpigmentation. Pituitary apoplexy occurred in one of these patients and another was operated upon because of visual field losses.

It is concluded that the remission obtained after adrenalectomy for Cushing's disease increases the survival rate and makes a normal life possible in most patients. However, persistent hypertension and cardiovascular disease as well as the possibility of the development of a pituitary tumour make the late prognosis somewhat uncertain.
MATHEMATICAL ANALYSIS FOR THE INTERPRETATION OF IN VIVO TRACER INFUSION EXPERIMENTS
Authors: Gurpide Erlio
Published: October 2009

In vivo tracer experiments involving intravenous infusion of 1, 2 or 3 labelled hormones are described in schematic form. Formulae are provided to calculate from steady state isotopic data, rates of production and interconversion of circulating compounds, foeto-maternal production and transfer of hormones and rates of metabolic reactions occurring in an organ. Formal procedures to convert these formulae to expressions applicable to single injection experiments are outlined. The simplicity of the mathematical basis for the interpretation of most current experiments in hormone research is emphasized.
EVALUATION OF MECHANICAL SYSTEMS USED IN PERFUSION
Authors: Bernstein Eugene F.
Published: October 2009

Among the critical factors in organ perfusion are (1) the mechanical components of the system, (2) the composition of the perfusate, and (3) the perfusing conditions. In this review, particular consideration is given to the pump, the oxygenator, and cannulas in such systems. Emphasis is placed upon the selection of pertinent equipment for the goals of a particular perfusion experiment, based upon the criteria of adequacy of the perfusion. Common problems in organ perfusion are summarized, and potential solutions to the perfusion problem, involving either biologic or mechanical extracorporeal systems, are suggested.
FUNCTIONAL ASSESSMENT OF AUTOTRANSPLANTED ENDOCRINE ORGANS
Authors: Goding J. R. , Baird D. T. , Cumming I. A. , McCracken J. A.
Published: October 2009

Autotransplants of the adrenal gland and of the ovary have been extensively used to study basal secretion and mechanisms of control of steroid secretions by these organs. The validity of the use of such preparations for this purpose is critically examined in this paper.

Autotransplantation did not materially affect the normal histological architecture of the glands. The adrenal became re-innervated and re-connected to the lymphatic system. Sheep with their remaining adrenal gland transplanted to the neck not only survived for many years but also could support normal pregnancy. The blood flow through the transplants was somewhat greater than that from the organs in situ. This was shown to be due to (a) absence of acute vasoconstriction due to recent surgery, (b) some degree of admixture with peripheral blood, (c) compensatory hypertrophy. Steroid secretion from the autotransplants was similar to that from in situ glands and could be increased by specific stimuli. Biosynthetic transformations, with simultaneous measurements of hormone secretion rates, can be studied under controlled conditions. Cyclic behaviour was disturbed when the ovary alone was transplanted to the neck, but this was restored when the ovary and the uterus were transplanted together to a cervical site.

Impairment of cyclicity was shown to result from the separation of the ovary from the uterus. The uterus was shown to affect luteal function by secreting the prostaglandin (PG) F2α. PGF2α gained access to the ovarian circulation by a countercurrent mechanism which was demonstrated to be quantitatively competent to cause luteolysis in the normal animal. The minimum infusion rate of PGF2α into the uterine vein of sheep on day 7 of the cycle which would cause luteolysis was about 20 μg/h. This is similar to the observed rate of PGF2α secretion in the uterine vein of ewes at the time of luteolysis. Hence PGF2α can be regarded as »the« luteolytic hormone in the sheep, and the ovine uterus as an endocrine organ.
PREPARATION AND MECHANICAL PERFUSION OF THE ISOLATED MONKEY BRAIN
Authors: White Robert J.
Published: October 2009

An isolated monkey brain model viably sustained by a mechanical extracorporeal perfusion system employing Dextran diluted subhuman primate blood has been developed. Each isolated brain is instrumented for continuously monitoring electrical activity (depth and cortical), ventricular pressure as well as intracerebral temperature and arterial (Circle of Willis) pressure. Employing EEG criteria the isolated brain approaches during the initial 60 to 90 minutes of perfusion the neurophysiological and metabolic performance of the intact brain. After the second hour of perfusion these preparations demonstrate accelerating electrical failure with an associated rise in excess lactate and a topographical cortical appearance consistent with developing cerebral oedema.

The solutions to problems of surgical isolation of the brain and design of extracorporeal perfusion support systems for this delicate organ as well as the performance of the isolated monkey brain are reviewed.
THE INFUSED TESTIS
Authors: Eik-Nes Kristen B.
Published: October 2009

The infused testis has been discussed as a model system for studies of processes leading to formation and secretion of testosterone. While this model possesses certain advantages over other systems used in such studies, it is far from perfect and a rather large battery of techniques in vivo and in vitro are required in order to obtain adequate information on steroidogenesis in the male gonad.
THE STRUCTURE OF EUKARYOTE CHROMATIN
Authors: Dounce A. L. , Chanda S. K. , Ickowicz R. , Volkman D. , Palermiti M. , Turk R.
Published: October 2009

In this paper certain aspects of the structure of chromatin are considered, with emphasis on the relationships of histone and residual protein to DNA. Recent findings relative to the complexing of histone with DNA are reviewed and material from this laboratory concerning the ratio of total histone to DNA and related topics are discussed. The questions of DNA strandedness and DNA repeating sequences are considered briefly. The relationship of residual protein of the cell nucleus to the structure of the chromosome is taken up and the bearing of the results of studies of nuclear gels on this problem are discussed. Recent results from this laboratory on the use of shear and a new autolytic procedure for detaching DNA from residual protein are outlined. The failure of residual protein to cause a marked elevation in the tm of nuclear DNA is discussed and material on the density gradient centrifugation of the DNA-residual protein complex is presented. Recent work on the fractionation of residual protein obtained from chromatin and isolated nuclei is critically reviewed. Finally a new hypothesis concerning a protein core structure in eukaryote chromosomes is presented briefly in the form of a model which may help to rationalize previously unexplained observations.
FUNCTIONAL STUDIES ON CHROMATIN
Authors: Paul J. , Carroll D. , Gilmour R. S. , More J. A. R. , Threlfall G. , Wilkie M. , Wilson S.
Published: October 2009

Functional studies on chromatin include assays of template activity and template capacity. Template activity is the measure of the rate of polynucleotide synthesis supported by a chromatin template in an incubation mixture including exogenous DNA-dependent RNA polymerase. It can be a highly misleading criterion unless great care is taken to standardise conditions. Template capacity is the measure of the DNA in a preparation of chromatin as compared with purified DNA, which is accessible to transcription by exogenous DNA-dependent RNA polymerase. It is determined by studying the complexity of the RNA made in the reaction, usually by RNA-DNA hybridization. The effects of different methods of preparation of chromatin, different RNA polymerases, conditions of incubation and purification of RNA are discussed.

The application of these and other methods to the study of reconstituted chromatin is described.
ON THE ROLE OF RNA IN GENE AMPLIFICATION
Authors: Brown Ronald D. , Mattoccia Emilio , Tocchini-Valentini Glauco P.
Published: October 2009

A complex of ribosomal DNA with RNA has been isolated from ovaries of metamorphosing tadpoles of Xenopus laevis. The complex is disrupted by treatments which destroy hydrogen bonds and the sedimentation of the DNA within the complex is sensitive to RNase. We suggest that the RNA: rDNA complex is an intermediate in the synthesis of amplified rDNA and that the role of the RNA in the process is that of a template. In addition, in a preliminary attempt to mimic the amplification process in vitro, we have demonstrated the use of this RNA from the complex as a template for DNA synthesis by a DNA polymerase isolated from ovaries of Xenopus.
RIBOSOME-MEMBRANE INTERACTION AND PROTEIN SYNTHESIS
Authors: Tata J. R.
Published: October 2009

The role of secretion of proteins for the attachment of ribosomes to membranes has been well established. That another function must exist for membrane-ribosome interaction is suggested by observations on: (a) the active synthesis of proteins on membrane-bound ribosomes of predominantly non-protein secreting cells, and b) the massive proliferation of membrane-bound ribosomes during active growth and development of both secretory and non-secretory tissues.

Literature on functional and compositional differences between membrane-bound and free ribosomes is reviewed and it is proposed that a major function of ribosome-membrane interaction is to effect a topological segregation of different populations of ribosomes synthesizing different classes of proteins.
OVALBUMIN mRNA, COMPLEMENTARY DNA AND HORMONE REGULATION IN CHICK OVIDUCT
Authors: Schimke Robert T. , Rhoads Robert E. , Palacios Raphael , Sullivan Drew
Published: October 2009

Chick oviduct differentiation is controlled by various steroid hormones, including both oestrogen and progesterone. Administration of oestrogen to chicks results in differentiation of tubular gland cells, which synthesize differentiated cell products, including ovalbumin and conalbumin. Ovalbumin can constitute as much as 60% of the protein synthesized by oviduct in the fully stimulated chick. Methods have been developed that allow for the quantitative assay of ovalbumin mRNA, the specific isolation of ovalbumin synthesizing polysomes by use of immunologic techniques, as well as the synthesis of a DNA probe complementary to the ovalbumin mRNA. With these techniques we have found that the rate of ovalbumin synthesis is proportional to the content of ovalbumin mRNA. Hybridization studies using the DNA probe indicate that there are only two copies of the ovalbumin gene per genome, and that there is no amplification of the ovalbumin genes to account for the large amount of ovalbumin synthesized.
TERATOGENIC SCREENING METHODS AND THEIR APPLICATION TO CONTRACEPTIVE PRODUCTS
Authors: Tuchmann-Duplessis H.
Published: October 2009

After an analysis of the basic principles of teratogenic drug testing and the complexity of factors involved in the production of congenital malformations, the following topics are considered:

the mechanisms of the teratogenic action

the role of the placenta in drug transfer

the choice of an animal model for teratogenic testing of hormonal sex steroids

the influence of oral contraceptives upon various phases of reproduction.

To compete with the various mechanisms which control the normal intrauterine development, multiple conditions have to be fulfilled. Such a situation occurs only occasionally in humans.

Much data are available but little knowledge exists regarding the teratogenic mechanisms and the precise role of the placenta in drug teratogenicity.

Great differences of reactions to drugs exist among various species. They can be related to differences in the enzymatic systems involved in the metabolic turnover of a drug, but in many cases no explanation is yet available.

It seems that preliminary screening of sex steroids could be done in rodents. This can contribute to the selection of interesting therapeutical agents and may aid in discovering potential side effects. Since the maintenance of pregnancy in rodents requires functioning ovaries, while in women and in primates the placenta takes over very early the hormonal control of pregnancy, it is suggested to use primates for the safety evaluation of contraceptive drugs. The results obtained in these species are likely to have good predictive value.

Oral contraceptives have no mutagenic effects, they do not produce chromosomal aberrations and seem to be devoid of teratogenic action.

Experimental and clinical data suggest that oral contraceptives have no adverse effects on the perinatal and postnatal development.
SPECIAL REQUIREMENTS FOR PLASTIC DEVICES
Authors: Bischoff Fritz , Bryson George
Published: October 2009

The safety testing for polymers in contraceptive devices already in use, at the human pilot test stage, or proposed is appraised. At the subcutaneous and intraperitoneal sites in the rat, polyethylene did not demonstrate chemical carcinogenicity; in the uterine horn endometritis was a prelude to epidermoid carcinoma development. In rodents, polyvinyl chloride copolymer, vinyl chloride, liquid silicone, and other polymers may respond as chemical carcinogens. A polyvinyl chloride resin produced local uterine squamous cell carcinomas in rats and leiomyosarcomas in mice. Intrauterine implants produce a characteristic foreign-body reaction in all species studied, including man, and secondary infection in rats and mice. Solid-state carcinogenesis can no longer be regarded as a phenomenon exclusive for rodents. Because it depends upon a minimum implant period in situ and a long latent period, its appearance in humans should occur only after many years and only if the implant has resided more than temporarily.

When safety testing fails because of differences in response between species and/or anatomical test sites, evaluation of the effects of polymers already used for other purposes in humans and over long periods of time may be a necessary recourse. The tendency of the silicone prosthesis to mechanically erode its way through body tissues and to absorb lipids suggest general lack of biological inertness.

The development of a fibrous capsule around parenteral implants may decrease metabolic exchange and limit the use of polymer-hormone depots.
SPECIAL PROBLEMS IN TOXICITY TESTING OF LONG ACTING DEPOT CONTRACEPTIVES
Authors: Neumann F. , von Berswordt-Wallrabe R. , Elger W. , Gräf K.-J. , Hasan S. H. , Mehring M. , Nishino Y. , Steinbeck H.
Published: October 2009

Two types of so-called "depot contraceptives", long-acting steroids which are of interest for human use, were studied in animals.

Norethisterone oenanthate, mainly gestagenic in the human and other species, turned out to be predominantly oestrogenic in rats. This oestrogenicity caused indirectly, via an enhanced hypophysial prolactin secretion, the well-known hypophysial and mammary tumours in rats.

Another synthetic gestagen, 4,6-dichloro- 17- acetoxy- 16α-methyl-4,6-pregnadiene-3,20-dione, which might be considered in its biological actions similar to preparations containing chlormadinone acetate or medroxy-progesterone acetate, induced no signs of oestrogenicity in dogs. It is surmised that its gestagenic influence indirectly, and probaby, via an enhanced hypophysial prolactin secretion caused "mammary nodules" in this "non-rodent" species. These studies have born out mainly two facts:

A synthetic steroid, norethisterone oenanthate, exerted different biological effects in different species: it was a gestagen in the rabbit, whereas in rats, its predominant influence was oestrogenic.

The hypophysial prolactin secretion was enhanced in various species by different mechanisms: in rats, the oestrogenicity caused an increased prolactin plasma level, whereas in dogs, a gestagen with obviously no inherent oestrogenicity, 4,6-dichloro-17-acetoxy-16α-methyl-4,6-pregnadiene-3,20-dione, converted the histological appearance of the anterior pituitary into a condition with a greatly increased number of eosinophils. This histological finding was interpreted as an indicator for a hypersecretion of prolactin.

Hence, animal work with "gestagens" has only limited predictive value with respect to their possible effects in the human species. Therefore, inflexible recommendations are not helpful in solving the safety problem of long-acting steroids which affect primarily reproductive processes.
SPECIAL REQUIREMENTS FOR TESTING POST-COITAL CONTRACEPTIVES
Authors: Lerner Leonard J.
Published: October 2009

Post-coital antifertility agents are a group of diverse chemical and pharmacological substances that have one activity in common, the prevention of or the termination of pregnancy. As with other classes of drugs, comprehensive toxicity evaluation must be performed in laboratory animals and in humans. Additional studies, however, should be undertaken to emphasize the influence of the clinical protocols on toxicity. Much toxicological information can be gained during broad-spectrum pharmacological testing and this information together with the biological activities can give direction for extended toxicity studies. Choice of animal species for these studies should be governed by parallelism with man phylogenetically, metabolically and physiologically, in response to hormones and drugs, in the metabolism and kinetics of compounds related to the drug in question, in specific areas related to reproduction and in other ways. It may take several different species to satisfy some of these requirements. The indicated use of the drug makes mandatory a study of foetal-toxicity and teratogenesis, and sub-antifertility doses as well as the recommended clinical dose should be employed in these studies. New types of drugs and new chemical entities require in-depth study, careful evaluation of effects, careful and constant clinical monitoring, and rapid dissemination of information about undesirable effects. While study of post-coital agents may not require great deviation from the manner in which other kinds of drugs are studied for toxicity, the fact that healthy women may be given such drugs for many years makes it imperative that all possible measures be taken to safeguard the health of these women.
SEX HORMONES – EFFECTS ON PROLACTIN CELLS IN THE RAT, DOG, MONKEY AND MAN
Authors: El Etreby M. F. , Günzel P. , Soulioti G.
Published: October 2009

Histomorphological and immunocytochemical methods were used to study the effect of either physiological and pathological endogenous production or long-term exogenous application of high doses of sex (steroid) hormones on the prolactin cells in the anterior pituitary of experimental animals and man.

Using these methods it was shown that enhanced endogenous secretion of sex hormones, mainly of oestrogens, seems to produce nearly the same morphological changes in the anterior pituitary of the rat, dog and man. It was also possible to show that long term exogenous application of high doses of oestrogens or oestrogens and progestagens can stimulate prolactin cells in some experimental animals.

The demonstration of similar mechanisms in man calls for more extensive investigations on the possible role of the sex hormones on prolactin secretion.
CHARACTERIZATION OF SPERMATOZOAL AUTO-, ISO- AND ALLO-ANTIGENS
Authors: Voisin G. A. , Toullet F. , Almeida M. D'
Published: October 2009

Three approaches are utilized to study and characterize spermatozoal antigens.

An immunological approach has demonstrated the presence of spermatozoal auto-, iso- and allo-antigens. Spermatozoal auto-antigens studied by several authors are able to induce the whole spectrum of immune reactions (delayed hypersensitivity, complement-fixing antibodies and anaphylactic antibodies) as well as of autoimmune aspermatogenic orchiepididymitis (AIAO). Different extraction procedures result in various preparations and even in different independent autoantigens (at least four), one protein, one membrane-linked antigen and at least two glycoproteins. Spermatozoal iso-antigens stricto sensu are determined by the Y chromosome and are present on at least 50 % of the spermatozoa. Spermatozoal allo-antigens are also present at the surface of spermatozoa, especially blood group antigens (ABO and MNS systems), transplantation antigens (HL-A, H-2) and also some other unidentified ones.

A biochemical approach has mainly been directed towards spermatozoal enzymes that have been shown to be antigenic even in the species of origin. This is the case for lactic dehydrogenase LDH-X (a mid-piece enzyme) and for acrosomal enzymes, e. g., hyaluronidase, possibly sorbitol dehydrogenase and trypsin-like acrosomal proteinase (the auto- and allo-antigenicity of the latter having not been established). At least three of these enzymes are known or supposed to play a role in the process of fertilization.

A clinical approach has described the presence of spermatozoal-coating antigen(s), such as transferrin or blood group substances from secretors obtained following the admixture of the secretions of the seminal vesicles. Indications were also obtained for the existence of antibodies directed against defined antigens. Several types of localization of antibodies on spermatozoa were described: acrosome (front part), equatorial segment, post-nuclear region, mid-piece and tail. Attempts at fractionation of human spermatozoal antigens are still at a preliminary stage.

Whatever the approach, the main interest of these antigens is that they are able to induce, in the species of origin or in a related species, antibodies capable of interfering with the normal process of reproduction, especially fertilization. In view of possible applications in the male or in the female, a few immunological considerations are of primary importance, particularly the localization of antigens (and their accessibility at the spermatozoal surface), their biological necessity for the reproductive process, the passage of the immune agents from the intravascular compartment to the genital tract, the possibility of inducing local immunization as well as to be able to direct the immune reaction at will toward the formation of immune agents responsible for the rejection reaction (sensitized cells, complement-fixing antibodies and complement) or the facilitation reaction (facilitating antibodies or suppressive cells).
IMMUNOLOGICAL INTERFERENCE WITH FERTILIZATION
Authors: Shivers C. Alex
Published: October 2009

Rabbit antisera against mouse and hamster ovaries and sheep antisera against rabbit ovary have been used to demonstrate ovary specific antigens in these three species of mammals. That one or more of the specific antigens are located in the zona pellucida which surrounds the egg was indicated in immunofluorescence studies and by the effects antibodies have on the zona. After exposure to the antibody a precipitate forms on the zonae of preovulatory, unfertilized and fertilized eggs and all embryonic stages up to the time when the zona is shed. The precipitate was always seen near the outer surface of the zona suggesting that the active sites of the antigen(s) are located in this region. Trypsin and pronase which usually dissolve the zona were ineffective on the precipitated outer portion but did dissolve the inner, unprecipitated region. Species- and tissue-specificity and chemical nature of the zona are discussed.

Important roles for the zona antigen(s) in a number of early events in reproduction were indicated by the effects antizona antibodies have on fertilization and implantation. Pretreatment of unfertilized hamster eggs with zona precipitating antibodies was most effective in blocking sperm attachment to the zona. When the antibody was added subsequent to in vitro insemination, but before the sperm had passed through the outer region of the zona, sperm penetration was blocked. Mouse embryos treated with zona precipitating antibody and cultured in vitro failed to escape from the zonae, implying that implantation could be inhibited by antibody. Passive immunization of female mice with antizona antibodies and precipitattion of the zonae of hamster embryos in transfer experiments reduced the number of implantation sites. The identification of zona antigens and the effects of zona precipitating antibodies on early reproductive events encourage further studies on the zona in reference to immunological control of fertility.
TECHNIQUES FOR ELICITING MUCOSAL IMMUNE RESPONSE
Authors: Waldman R. H. , Ganguly R.
Published: October 2009

Development of techniques for eliciting an immune response on mucosal surfaces is a relatively new area of clinical research. With the recognition of the existence of secretory immunity, independent of the systemic system, there was renewed interest in re-examining the conventional approach for optimal immunization techniques. A large body of data indicate that the majority of the secretory immunoglobulins and antibody produced to antigenic stimulation of mucosal surfaces is locally produced. Thus, antibody to C. albicans in the cervical or vaginal mucus has been shown to be of local origin and it predominantly belongs to the secretory IgA immunoglobulin class. The mechanism of antigen processing by the secretory surface leading to antibody formation remains a mystery, but it might be determined by the selective localization of antigens in the reticuloendothelial cells of the lamina propria, bronchi or small intestine. Usually application of antigen topically to the mucosal surface elicits local antibody formation to a greater extent than does parenteral immunization. On the other hand, a more pronounced systemic immune response is seen when the antigen is administered systemically. However, a number of other factors determine the quality and quantity of the immune response, e. g., the physical state of the antigen, live vs killed vaccine, dose, adjuvant, previous exposure to similar or cross-reacting antigens, and site of application of the antigen. These factors are discussed in the review. Recent observations suggest that cell mediated immunity is a component of the secretory immune system, and like the humoral mechanism, also may be partially compartmentalized.
SURFACE ANTIGENS OF NORMAL EARLY EMBRYOS AND A TUMOUR MODEL SYSTEM USEFUL FOR THEIR FURTHER STUDY
Authors: Edidin Michael , Gooding Linda R. , Johnson Martin
Published: October 2009

We review experiments on the presence of strong transplantation antigens, H-2 antigens, in mouse gametes and early embryos. Parallels are drawn between antigenic structure of embryos and tumour cells, and experiments with a model tumour system, a teratoma, are presented indicating the presence in early embryos of surface antigens which are shared with the tumour.
I. ANTI-ANDROGENIC EFFECTS OF A NEW STEROID TSAA-291 (16β-ETHYL-17β-HYDROXY-4-OESTREN-3-ONE) AND ITS DERIVATIVES
Authors: Nakayama Ryo , Masuoka Michio , Masaki Tsuneo , Shimamoto Kiro
Published: October 2009

Anti-androgenic properties of newly synthesized steroidal compounds, TSAA-291* and its derivatives, were studied in the rat.

(1) Subcutaneous administrations of TSAA-291 and TSAA-272* were effective in depressing the weight of seminal vesicles and ventral and dorsal prostates in the adult rat. (2) TSAA-291, TSAA-328* and TSAA-272 were shown to be anti-androgenic by the subcutaneous or oral route in the immature castrated male rat treated with testosterone propionate. The dose-response curves of all the test agents expressed in terms of percentage inhibition were approximately parallel to each other. Cyproterone acetate depressed the weight of levator ani muscle dose-dependently, whereas TSAA-291, TSAA-328 and TSAA-330* did not inhibit the muscle weight in a dose range sufficiently effective in inhibiting the weight of seminal vesicles and ventral and dorsal prostates. (3) TSAA-291 was also antiandrogenic against androstenedione and dehydroepiandrosterone.

(4) Local administration of the anti-androgenic compounds into the left seminal vesicle or left testis of the young adult rat depressed the weight of the seminal vesicle or testis on the treated side as compared with the vehicle-injected contra-lateral side.
III. HORMONAL SPECTRA OF ANTI-ANDROGEN TSAA-291 (16β-ETHYL-17β-HYDROXY-4-OESTREN-3-ONE) AND ITS DERIVATIVES
Authors: Masuoka Michio , Masaki Tsuneo , Yamazaki Iwao , Hori Tatsuhiko , Nakayama Ryo
Published: October 2009

For the purpose of obtaining hormonal spectra of anti-androgen TSAA-291* and its derivatives, a variety of endocrine characteristics were studied.

(1) Androgenic and anabolic activity: Subcutaneous administration of anti-androgen TSAA-291 and its acetate, TSAA-328*, to the immature orchiectomized rat resulted in significant weight increase of the levator ani** but in only a nominal response of seminal vesicles and prostates even at a large daily dose of 9.6 mg. The resultant anabolic/androgenic ratio was estimated to be extremely high. (2) Oestrogenic activity: Uterine weight in response to these anti-androgens were sluggishly dose-dependent, and the maximal plateau response remained considerably lower than that induced by oestradiol-17β. The oestrogenic activity of these anti-androgens was estimated to be 1/200 000 or less as that of oestradiol-17β. A single subcutaneous dose of 100 mg of TSAA-291 or its caproate, TSAA-330*, did not induce the vaginal cornification in the adult ovariectomized rat. (3) Anti-oestrogenic activity: Antagonistic effect of these anti-androgenic compounds on the uterine weight response to oestradiol-17β was found in the immature ovariectomized rat. A single subcutaneous dose of 100 mg of TSAA-291 or TSAA-330 also induced the antagonism against the cornification caused by daily treatments with 1 μg oestrone in the adult ovariectomized rat. (4) Progestational activity: These anti-androgenic compounds proved to be less active than progesterone in the McPhail's test. (5) Anti-inflammatory activity: Daily subcutaneous dose of 20 mg of TSAA-291 for 6 days did not significantly depress the weight of granuloma developed around the cotton-pellet implanted in the young male rat. TSAA-291 did not affect the anti-inflammatory action of 1/6 mg of prednisolone phosphate. Combination of both agents seemed to be effective in enhancing the anti-androgenic action of TSAA-291, whereas prednisolone phosphate alone rather increased the weight of the accessory sex organs. (6) Liver glycogen deposition activity: Daily intramuscular doses up to 38.4 mg of TSAA-291 for 5 days did not increase the liver glycogen level in the adrenalectomized rat.
VII. ON THE MECHANISM OF ANTI-ANDROGENIC ACTION OF 16β-ETHYL-17β-HYDROXY-4-OESTREN-3-ONE (TSAA-291)
Authors: Nakayama Ryo , Masuoka Michio , Hiraga Kentaro , Miki Takuichi
Published: October 2009

The mechanism of anti-androgenic action of a steroidal compound, TSAA-291, was summarized and discussed in reference to its drug-designing and structure-activity relationship. The target of the drug-design was to obtain a substance which is inactive in androgenic activity and is capable of antagonistically competing with androgen for the receptor. With this intention, the androgen molecule was rendered with a steric hindrance influencing upon the functional 17β-hydroxy group. Introduction of a bulky group at the steroidal position-13 or -16 led to anti-androgenic properties. Intense steric hindrance by introducing an enormously bulky group or complete elimination of the 17β-hydroxy group rather decreased the anti-androgenic activity. Of these anti-androgens thus synthesized, TSAA-291 proved to be the most active in the anti-androgen assay and also antagonistic against the uptake of [3H]testosterone by the rat ventral prostate.
Prevalence and Incidence of Type II Diabetes in Denmark Compared with Other Countries
Authors: Damsgaard Else Marie
Published: October 2009

ABSTRACT. The prevalence of type II diabetes in Denmark increases with age to a maximum about the age of 70 years and the increase is higher for known diabetics than for previously undiagnosed diabetics. In the age group 60–74 years the total prevalence of diabetes was 6.1 % (5.5 % for women and 6.6 % for men).

The results of glucagon tests seem to indicate that about 60 % of insulin treated subjects after the age of 60 years are of type II. All diet treated diabetics and nearly all tablet treated and newly diagnosed diabetics are of type II.

The incidence of type II diabetes in Denmark has not been well described and might only be determined by reexamining the same fixed population with short intervals.

Differences in ascertainment, methods, and diagnostic criteria, from one study to another make a close comparison of different studies impossible.

Key words: prevalence, incidence, epidemiology, maturity onset diabetes.