bluuebunny/crossref_metadata_2025 huggingface dataset:
PRINCIPLES FOR THE ASSESSMENT OF THE RELIABILITY OF RADIOIMMUNOASSAY METHODS (PRECISION, ACCURACY, SENSITIVITY, SPECIFICITY)
Authors: Midgley A. Rees , Niswender Gordon D. , Rebar Robert W.
Published: October 2009

This paper represents an attempt to formulate working definitions and principles by which the reliability of a given radioimmunoassay can be evaluated and compared with other radioimmunoassays. Statistical methods for the analysis of radioimmunoassay data are presented. The feasibility of utilizing the logit transformation has been tested empirically, and the advantages which accrue from analyzing data in this fashion are considered. The statistical transformations take into account the heteroscedasticity of the error inherent in a radioimmunoassay and the non-homogeneity extant in the confidence intervals about the estimates of the values of individual samples. By defining sensitivity as the smallest quantity of unlabeled hormone which can be measured with statistical certainty, and precision as the error in the determination of a hormone concentration at each particular point along the standard inhibition curve, it becomes possible to compare established assays in a meaningful fashion. Minimal criteria for establishing specificity, here regarded as freedom from interference by unwanted substances, are presented. Problems in the evaluation of relative and absolute accuracy are also discussed.
RADIOIMMUNOASSAY OF FOLLICLE-STIMULATING (FSH) AND LUTEINIZING (LH) HORMONES BY CHROMATOELECTROPHORESIS
Authors: Saxena Brij B. , Leyendecker Gerhard , Chen Weiyu , Gandy Hortense M. , Peterson Ralph E.
Published: October 2009

Procedure for the radioimmunoassay of FSH and LH using chromatoelectrophoresis for the separation of antibody-bound and free labeled hormone is described. Diurnal variation in FSH and LH levels of normal men and women in follicular and luteal phases of the cycle is reported. The diurnal variation was not abolished in three women taking contraceptive medication and in two women in postmenopause. The diurnal variation of FSH and LH in plasma is compared with that of cortisol, dehydroepiandrosterone and testosterone. It is suggested that diurnal variation of FSH and LH may be independent of a normally functioning pituitary-gonadal axis; however, steroidal feed-back may have a regulatory effect on the magnitude of the diurnal variation. The plasma levels of FSH and LH in normal subjects during normal menstrual cycles as well as in patients with endocrinopathies obtained in our laboratory by radioimmunoassay are compared with those obtained by both radioimmunoassay and bioassay by other investigators.
RADIOIMMUNOASSAY EMPLOYING POLYMERIZED ANTISERA
Authors: Donini S. , Donini P.
Published: October 2009

Antisera made insoluble by polymerization were used for the radioimmunoassay of FSH and LH, as this was shown to be a useful method of separating bound and free labelled hormone.

Highly purified HCG and FSH, prepared in this laboratory, from urinary source were labelled with 125I. Antisera were raised by injecting urinary materials. The specificity of antisera was studied and some cross-reactivity was found between anti-urinary FSH and LH, but not with HCG. The sensitivity of LH and FSH radioimmunoassay was 1 mIU/ml and 3 mIU/ml, respectively.

Some determinations in dialyzed urine from various sources have also been performed and the preliminary results obtained are compared to those previously reported by others.

This paper discusses the radioimmunoassay of FSH and LH using a new solid-phase method, i. e. the use of polymerized antiserum, as previously described (Donini et al. 1968).
CHEMICAL PRECIPITATION TECHNIQUES FOR THE RADIOIMMUNOLOGIC QUANTITATION OF GONADOTROPHINS WITH SPECIAL REFERENCE TO THE ORGANIC SOLVENT DIOXANE
Authors: Thomas K. , Nash D. , Ferin J.
Published: October 2009

A brief survey of the literature reveals that, for the radioimmunoassay of hormones, separation of free from antibody-bound hormone has been accomplished by increasing salt concentration (salting-out), precipitation with organic solvents (primarily ethanol) or combinations of both. The application of these techniques has been most successful with hormones in a relatively low protein milieu such as urine or buffers (standard curve analysis) but are somewhat limited for use on high protein mixtures such as plasma or serum.

A detailed account of a technique involving precipitation of hormoneantibody complexes with the organic solvent dioxane is given. This technique has been most successfully applied to the quantitation of luteinizing hormone (LH) in plasma as well as in urine.

A discussion of the problems encountered due to the specificity of the antibody and purity of tracer antigens (hormones), in the radioimmunoassay procedure for luteinizing hormone, is included.
A COMPARISON OF BIOLOGIC AND IMMUNOLOGIC POTENCY ESTIMATES OF HUMAN LUTEINIZING (LH) AND FOLLICLE STIMULATING (FSH) HORMONES
Authors: Ryan Robert J.
Published: October 2009

Published comparisons of biologic and immunologic potency estimates of human pituitary and urinary FSH and LH have been examined. In general, when pituitary fractions were assayed against pituitary standards and urinary fractions against urinary standards, there was good agreement between the two assay systems for both LH and FSH. However, when pituitary fractions were assayed against urinary standards, and vice versa, disparity occurred. This disparity was greater for LH than for FSH and was of such a nature as to suggest that the urinary hormone lost immunologic potency to a greater degree than biologic potency. A hypothesis, based on the formation of hybrid molecules, is presented to explain this immunologic deficiency of urinary LH. Evidence is also given to indicate that a dissociated, biologically inactive but immunologically recognizable form of LH, as well as the biologically active associated species of LH, is excreted in urine.
THE CORRELATION BETWEEN IMMUNOLOGICAL AND BIOLOGICAL ESTIMATION OF HCG IN BODY FLUIDS
Authors: Wilde C. E.
Published: October 2009

Biological estimates of HCG do not always agree with those obtained by immunoassay. Possible reasons for this discrepancy including factors influencing specificity in both immunological and biological systems are enumerated. Evidence for the occurrence of antigens with similar immunological properties but varying biological potency is presented and the nature of the antigens particularly with relation to their sialic acid content and their effect on HCG assay is discussed.

Antigens of varying biological potency are present in biological standards and the nature of the standard as well as the characteristics of the antiserum will affect the results obtained in immunoassay.

A variety of bioassays are employed for the assay of HCG and these do not always give identical results and may not measure the same activities. The biological activity of HCG is therefore not easy to define and hormonal polymorphism and polymerisation and the synergistic and inhibitory effect of physiological fluids may modify biological activity.

It is suggested that HCG activity and the biological and immunological systems used for its quantitation should be better defined before agreement within methods and between methods can be improved. Meanwhile the deficiencies of both biological assay and immunological assay should be borne in mind and the technique chosen should be related to the purpose for which the estimates are required.
LABORATORY TESTS OF THYROID FUNCTION IN HYPERTHYROIDISM I.
Authors: Lamberg B.-A. , Heinonen O. P. , Viherkoski M. , Aro A. , Liewendahl K. , Kvist G. , Laitinen O. , Knekt P.
Published: October 2009

For the elaboration of a computer programme for the diagnosis of thyroid diseases the pattern of the results of laboratory tests was studied after dividing 125 hyperthyroid patients into different clinical entities and further also according to the severity of the hyperfunction of the thyroid gland. Comparison of variables was made between different clinical entities and, within these, between different grades of severity. All these groups were compared with the controls. Furthermore, the correlation between the results of laboratory tests was studied. The patients were divided into the following groups: toxic diffuse goitre (TDG; 50 cases), toxic multinodular goitre (TNG; 52 cases) and toxic adenoma (TA; 23 cases), and according to clinical severity of hyperfunction of the thyroid gland, into the following grades: »borderline» (I; 16 cases), mild (II; 35 cases) and moderate to severe (III; 74 cases). The laboratory tests studied were: protein-bound iodine in the serum (PBI), uptake of triiodothyronine by Sephadex (T3U), »free thyroxine index» (T3I), free proportionate and absolute thyroxine (PFT4, AFT4), serum cholesterol, serum and urinary hydroxyproline (HOP), glucose-6-phosphate dehydrogenase activity in red blood cells (G-6-PD), 2.5 h and 24 h thyroid uptake of radioiodine, 24 and 48 h urinary excretion, protein-bound radioactive iodine in the blood (PBRI) and the 24 h conversion ratio (CR). There was a statistically significant difference between TDG and TNG with regard to the thyroid uptake of radioiodine and the free thyroxine, the mean values being lower in the latter group of patients. The TA group differed significantly from the combined TDG-TNG group, the mean values deviating towards the normal range. The TA group did not differ from the controls with regard to PFT4, cholesterol and E0–24. This group evidently represented a separate population.

In the TDG and TNG groups the laboratory data correlated well with the clinical assessment of severity. In most cases the differences were statistically significant. Grade I, however, differed from the controls only with respect to the variables of the hormone concentration in the blood.

The variables of the measurement of hormone concentration in the blood had a strong linear correlation (r = 0.6—0.9) to each other and to the 2.5 h uptake. A significant correlation (r = 0.5—0.6) was also found between the variables mentioned and the urinary HOP and G-6-PD. The correlation between HOP and G-6-PD was of the same order.

The fact that the pattern of PFT4—AFT4 and T3U—T3I differed may indicate variations in the ratios between triiodothyronine and thyroxine in different clinical entities of hyperthyroidism.
LABORATORY TESTS OF THYROID FUNCTION IN HYPERTHYROIDISM II.
Authors: Lamberg B.-A. , Heinonen O. P. , Liewendahl K. , Kvist G. , Viherkoski M. , Aro A. , Laitinen O. , Knekt P.
Published: October 2009

The distributions of 13 variables based on 10 laboratory tests measuring thyroid function were studied in euthyroid controls and in patients with toxic diffuse or toxic multinodular goitre. Density functions were fitted to the empirical data and the goodness of fit was evaluated by the use of the χ2-test. In a few instances there was a significant difference but the material available was in some respects too small to allow a very accurate estimation. The normal limits for each variable was defined by the 2.5 and 97.5 percentiles. It appears that in some instances these limits are too rigorous from the practical point of view. It is emphasized that the crossing point of the functions for euthyroid controls and hyperthyroid patients may be a better limit to use. In a preliminary analysis of the diagnostic efficiency the variables of total or free hormone concentration in the blood proved clearily superior to all other variables.
ZUR BEDEUTUNG DES HYPOPHYSÄREN LUTEINISIERUNGSHORMONS FÜR SCHWANGERSCHAFT, GEBURT UND LACTATION BEI DER RATTE
Authors: Loewit K.
Published: October 2009

The role of luteinizing hormone (LH) for the maintenance of pregnancy, parturition and lactation was investigated by immunological and histochemical methods in the rat. Neutralisation of endogenous rat-LH with Rabbit-Anti-Bovine-LH-Serum (selective hypophysectomy) from days 7-12 of pregnancy resulted in reabsorption of the foetuses and the reappearance of strong 20α-hydroxysteroid-dehydrogenase (20α-OHSD) activity in the corpora lutea (CL) of pregnancy, which normally show no such activity at that time. This effect could be prevented in part by concurrent pregnenolone administration and fully by progesterone, but was not influenced by oestrogen or prolactin. It is concluded that in early pregnancy LH is the main luteotrophic hormone in the rat even though prolactin might act synergistically with it. Antiserum treatment after the 12th day of gestation had no influence on the state or duration of pregnancy or on parturition. LH-injections during the first half of pregnancy had no luteolytic effects i. e. they did not activate 20α-OHSD activity. After day 16 they advanced the reappearance of the enzyme, but delayed parturition or resulted in stillbirths. Neither LH nor antiserum seemed to alter lactation. Since progesterone prevented both the termination of pregnancy and the recurrence of 20α-OHSD activity, it should have some regulatory properties on the enzyme. It is discussed whether the gonadotrophin-dependent progesterone level could regulate the 20α-OHSD activity rather than result from it.
PREPARATION AND CHARACTERISTICS OF CORTICOSTEROID-BINDING GLOBULIN (CBG, TRANSCORTIN)
Authors: Westphal Ulrich
Published: October 2009

The corticosteroid-binding globulin (CBG, transcortin) rises in the blood of the developing mammal from small concentrations in the young to adult levels which are similar in man and woman, but twice as high in the female rat as in the male. The affinity of the CBG complexes with corticosteroids is maximal at approximately pH 8; it decreases with increasing temperature. Study of the relative binding strength for different steroids clearly indicated a species specificity of the CBG molecules. This was confirmed by the isolation of CBG from serum of man, rat and rabbit, accomplished by a variety of chromatographic procedures. The pure glycoproteins, homogeneous by physicochemical criteria, proved to be distinct molecules on the basis of amino acid and carbohydrate composition and other properties. They possess one principal steroid binding site. Thermodynamic data indicate a very tight fit of the interacting components in the complexes. Removal of about 90% of corticosterone from its complex with rat CBG by gel filtration at 23° results in polymerization of the glycoprotein molecule to dimeric, tetrameric and octameric forms. Recombination of the polymeric mixture with 1 mole corticosterone per 53 000 g CBG re-forms the monomer. Possible biological significance of this reversible polymerization of a carrier protein under the regulatory control of the binding steroid hormone has to be explored.
FURTHER ASSESSMENT OF THE RELIABILITY OF PROGESTERONE ASSAYS BY COMPETITIVE PROTEIN BINDING
Authors: Souza M. L. A. de , Williamson H. O. , Moody L. O. , Diczfalusy E.
Published: October 2009

Experimental variables influencing the determination of plasma progesterone by competitive protein binding analysis have been further assessed. The precision of the method has been improved by additional mechanisation. Method interfering factors introduced by the extraction and purification procedure have been evaluated quantitatively. Another type of interfering material is present in plasma extracts from which all progesterone has been removed. The displacement effect of this material is additive to that derived from solvent and chromagram residues.

Unless the various types of interfering material are taken into proper consideration, competitive protein binding methods for steroids may yield erroneous results.
TESTOSTERONE ASSAYS BY COMPETITIVE PROTEIN BINDING
Authors: Vermeulen Alex , Verdonck L.
Published: October 2009

A review of competitive protein binding methods for the determination of testosterone in plasma is given.

The different steps discussed are: Choice of the binding protein, dilution of the protein solution, separation of the free from the bound fraction, purification of the extract and factors determining the blank.

The method used in the author's laboratory is critically discussed. The recovery is 98% ± 4% at 1 ng. The coefficient of variation is 3% at 660 ng/100 ml, 13% at 35 ng/100 ml, and 40% at 20 ng/100 ml. The sensitivity is about 10 ng/100 ml.
ASSAY OF ALDOSTERONE BY COMPETITIVE PROTEIN BINDING
Authors: Robinson Ralph G. , Fanestil Darrell D.
Published: October 2009

We have applied a competitive protein binding technique to the assay of aldosterone, utilizing the aldosterone binding protein (ABP) in kidney as the specific receptor molecule. Intracellular ABP was obtained from the homogenization of adrenalectomized rat kidneys in 0.25 M sucrose. The 10 000 × g supernatant contained the greatest binding affinity for aldosterone and was used without further modification. 1,2-3H-aldosterone was used as the tracer. After maximum binding, which occurred after 5 hours of incubation at 0–4°C, the aldosterone-ABP complex was stable for several hours and could be separated from unbound aldosterone by gel filtration using a 1 × 15 cm Biogel P-10® column. However, since the aldosterone-ABP complex was relatively easily dissociated by warm temperatures, all procedures had to be conducted at 4°C. Competitive inhibition of aldosterone binding to ABP occurred with 9-alpha fluorocortisol, cortisol, and progesterone. No inhibition of binding was found with oestradiol. Using this in vitro binding system, 1.05 × 10−12 and 2.10 × 10–12 mol aldosterone were assayed with coefficients of variation of 13.4% and 13.3%, respectively. Because cortisol effectively competes with aldosterone for ABP, application of this technique to biological fluids must be preceded by separation of aldosterone from cortisol. However, when this separation was achieved with one-step descending paper chromatography, our blank value was high and variable. This is apparently because some material which eluted from the paper interfered with the association of aldosterone and ABP. Therefore, this sensitive assay for aldosterone cannot be applied to biological specimens until the problem of the non-specific blank is circumvented.
VIRUS ASSISTED FUSION OF EMBRYONIC CELLS
Authors: Graham Christopher F.
Published: October 2009

A method is described for fusing mouse blastomeres together. These fused combinations develop into tetraploid blastocysts. On subsequent transfer to a foster mother they develop into large trophoblast masses.
IMMUNOHISTOCHEMISTRY OF INDIVIDUAL ADENOHYPOPHYSIAL CELLS
Authors: Leleux P. , Robyn C.
Published: October 2009

Immunohistochemistry is the intracellular detection of antigens by the use of specific antibodies labelled with a tracer. The choice of the tracer is such that the sites of the antigen-antibody reactions can be visualized by microscopic examination. The present report refers to the human pituitary where most of the immunohistochemical identifications of adenohypophysial cells were conducted with antibodies specific of their hormonal content and labelled with fluorescein isothiocyanate as tracer. Such immunohistochemical identifications had to be correlated to the morphological nomenclatures of the glandular cells based on histochemical stainings. Confusion has been introduced in these nomenclatures by the definition of three to eight cell types using different criteria and different terminologies. In the present report, owing to this absence of standardization, the comparative evaluation of immunohistochemical data have been based on Romeis (1940) and Pearse & van Noorden (1963) nomenclatures.

There is strong experimental evidence supporting the localization of adrenocorticotrophic hormone (ACTH) in the basophils of Romeis β type (R-mucoids of Pearse). Somatotrophic hormone (STH) has been consistently found in the acidophils of Romeis a type. In the human, there is no direct evidence to support the localization of prolactin (LTH) in the acidophils of Romeis ε type. Luteinizing hormone (LH) and follicle stimulating hormone (FSH) have both been located in the basophils of Romeis δ type (S-mucoids of Pearse). Further investigations into the human and also into other mammalian species are required to determine if the gonadotrophic hormones have different localizations on the cellular or on the subcellular level. The immunohistochemical localizations of thyroid stimulating hormone (TSH) and melanocyte stimulating hormone (MSH) have not been convincingly achieved.

The conclusions drawn from immunohistochemical studies of the adenohypophysis are essentially limited by the cross reactions existing between STH and prolactin, between ACTH and MSH and between LH, FSH and TSH.

More experimental data on these immunological cross reactions are still required before more accurate morphological discriminations can be achieved between the cell types secreting STH and prolactin, between those secreting ACTH and MSH and between those secreting LH, FSH and TSH. In addition, when hormones of the adenohypophysis are chemically and/or antigenically closely related, the cells responsible for their secretion are morphologically very similar too.

Finally, immunohistochemical studies revealed the lack of species specificity of the pituitary hormones. Extensive cross reactions have been shown between human STH and STH of all mammalian species studied so far. Consistent cross reactions were also found between human gonadotrophins and those of several mammalian species.
ASSESSMENT OF SEX STEROID ACTION IN VITRO
Authors: Means Anthony R. , O'Malley Bert W.
Published: October 2009

This review summarizes the present knowledge concerning in vitro effects of sex steroid hormones. Moreover, methods are described which may allow the assessment of oestrogen action on chick oviduct in vivo by assaying the cell free synthesis of a tissue specific protein (ovalbumin) in vitro.
IN VITRO ASSAYS OF ADENYL CYCLASE
Authors: Rodbell Martin
Published: October 2009

A brief review of the properties and current knowledge of the components of adenyl cyclase systems in animal cells is presented, followed by some general remarks on the problems of assaying adenyl cyclase. Recent techniques developed for the assay of adenyl cyclase in broken cell preparations are described. Methods which determine the levels of cyclic 3′5′AMP in intact cells or tissues are also presented, with particular emphasis on the use of binding of the nucleotide to cyclic 3′5′AMP dependent protein kinases and radioimmunoassays of the cyclic nucleotide.
PLACENTAL PERFUSION IN VITRO
Authors: Cedard L.
Published: October 2009

Because of its anatomical situation, easily accessible vascular pedicle and closed foetal circulatory system, the chorioallantoic placenta is particularly suitable for perfusion. It is possible to perfuse the whole organ or a limited part of it, such as a single cotyledon or a group of cotyledons. Perfusion methods have been employed for studying some of the numerous functions of the placenta. Such methods permit the study of the metabolism of the isolated organ as well as that of placental permeability to different metabolites, respiratory gases and drugs. Perfusion methods also make possible the investigation of placental endocrine function and seem to represent a more physiological approach than does the study of homogenates, slices or subcellular preparations.
TRANSPORT OF SUBSTANCES ACROSS PERFUSED ORGANS
Authors: Dancis J.
Published: October 2009

The subject of transport of substances through perfused organs has been briefly reviewed, stressing methodology rather than results. The emphasis has been on studies of transport across the placenta.
PROSTAGLANDIN E2: ANALYSIS OF EFFECTS ON PREGNANCY AND CORPUS LUTEUM IN HAMSTERS AND RATS
Authors: Labhsetwar A. P.
Published: October 2009

Effects of prostaglandin (PG) E2 on some reproductive processes of hamsters and rats were studied. The PG exerted antifertility effects in either species although it was 4–10 times less potent than PGF2α. Antifertility doses of PGE2 caused morphological degeneration of corpora lutea, induced fresh ovulations during the course of the treatment, significantly decreased peripheral progesterone levels and inhibited development of deciduomata in response to trauma. Both pregnancy and decidual growth could be maintained by simultaneous administration of exogenous progesterone. PGE2 also decreased plasma progesterone concentration in pseudopregnant-hysterectomized hamsters although the decrease was not significant when compared with intact pregnant or pseudopregnant hamsters. PGE2 produced no demonstrable effects on egg transport in hamsters but may have induced delayed implantation in rats by causing tubal retention of zygotes. The antifertility doses of PGE2 but not of PGF2α inhibited implantation of a proportion of blastocysts as studied by the Pontamine blue reaction and adversely affected the spacing mechanism of blastocysts in the uterus significantly reducing the distance between the adjacent implantation sites. When PGs E2 and F2α were given together to pregnant rats in doses which were virtually ineffective individually, the combined treatment caused loss of pregnancy. Administration of PGE2 to near term hamsters produced premature littering but oxytocin in doses tried proved inactive. Antifertility doses of PGE2 produced diarrhoea in rats but not in hamsters.

The results demonstrate that PGE2 shares the luteolytic property of PGF2α but in addition possesses several other properties not known to be shared by PGF2α.
ULTRASTRUCTURAL LOCALIZATION OF PROTEINS AND NUCLEOPROTEINS IN THE INTERPHASE NUCLEUS
Authors: Bouteille Michel
Published: October 2009

A review is given on the present state of knowledge of the location of diffuse and »figured« proteins, and ribonucleoproteins in the interphase cell nucleus. Attention is focused on structures of still obscure function, and on various types of nuclear inclusions which do not seem to be constant, but are presumably related to the degree of cell activity, whether physiologic or pathologic.
ISOLATION AND COMPOSITION OF NUCLEI AND NUCLEOLI
Authors: Busch Harris , Choi Yong C. , Daskal Ierachmiel , Inagaki Akira , Olson Mark O. J. , Reddy Ramachandra , Ro-Choi Tae Suk , Shibata Hirotoshi , Yeoman Lynn C.
Published: October 2009

The past decade has brought about the evolution of increasingly precise methods for isolation of nuclei and nucleoli and their components including histones, acidic proteins, high molecular weight RNA, low molecular weight RNA and ribonucleoproteins. The available methods and improving methods permit the isolation of individual unique molecular species in high states of purity. Elegant studies have been made on the amino acid sequences of some of the histones and on the nucleotide sequences of some of the species of low molecular weight RNAs uniquely found in the nucleus. In addition, nucleotide sequences are being defined for fragments of high molecular weight nuclear RNA. In studies on the effects of hormones on cells, the roles of repressor and derepressor acidic nuclear proteins are now being defined. The next decade should bring extensive new information regarding nuclear structures and functions and nucleolar components and their structures and functions.
INDUCTION OF SPECIFIC TRANSLATABLE MESSENGER RNA'S BY OESTROGEN AND PROGESTERONE
Authors: O'Malley Bert W. , Rosenfeld Gary C. , Comstock John P. , Means Anthony R.
Published: October 2009

In this manuscript, we report the de novo synthesis of two specific chick proteins, ovalbumin and avidin, in a heterologous rabbit reticulocyte lysate in vitro system. Synthesis of these proteins is absolutely dependent on RNA extracted from oviduct and not RNA extracted from non-target tissues. Furthermore, we have demonstrated that the intracellular accumulations of ovalbumin mRNA and avidin mRNA are directly dependent upon prior stimulation by oestrogen and progesterone, respectively.
FRACTIONATION OF MAMMALIAN CHROMATIN
Authors: Janowski Michel , Nasser DeLill S. , McCarthy Brian J.
Published: October 2009

Sheared chromatin may be separated into two peaks representing active and inactive fractions by gel exclusion chromatography on agarose columns. The active fraction is identified by the presence of nascent RNA chains synthesized in vivo or in vitro. Similarly, 3H labelled E. coli polymerase binds preferentially to the front active peak. Labelled hormones become associated with the chromatin of target tissues. Again this association occurs predominantly with the minor chromatin fraction identified as active by the above criteria. The basis for fractionation and the implications for our understanding of chromatin structure are discussed.
NUCLEIC ACID-REACTIVE ANTIBODIES SPECIFIC FOR NUCLEOSIDES AND NUCLEOTIDES
Authors: Erlanger B. F. , Senitzer D. , Miller O. J. , Beiser S. M.
Published: October 2009

A method is described for making anti-purine and anti-pyrimidine anti-bodies by immunization with conjugates of naturally-occurring nucleosides or nucleotides with carrier proteins. The specificities of the antisera are presented and shown to be predominantly, if not solely, for the determinant purine or pyrimidine group presented in the antigen. The antisera react with single-stranded or denatured DNA and, in the case of anti-adenosine, with RNA preparations. The antisera have been utilized in biochemical systems to inhibit the priming ability of DNA in a DNA-dependent DNA polymerase system, to detect minor bases such as 6-methyladenosine in DNA, and to detect small areas of single-strandedness in otherwise native DNA. The sera have also been shown to react with living cells, inhibiting the development of fertilized sea urchin eggs and entering and inhibiting transformed Chinese hamster lung cells without affecting normal ones. They have been used in immunofluorescence experiments in which they have been shown to react with nuclear DNA of fixed mouse L-cells harvested during the S phase. Moreover, they have found application in the characterization and identification of human and mouse chromosomes. Finally, initial studies have shown that they can be used in highly specific radioimmunoassays for purine and pyrimidine nucleosides.
CONTROL OF EXPRESSION OF GENETIC INFORMATION IN E. COLI BY CYCLIC AMP
Authors: Pastan Ira , Crombrugghe Benoit de
Published: October 2009

Cyclic AMP is required for the expression of a large variety of dispensable gene functions in E. coli. Cyclic AMP acts at the gene level to permit transcription of genes under its control. To do this cyclic AMP first combines with a cyclic AMP receptor protein and the cAMP-CRP complex alters the DNA so that RNA polymerase can bind in a form ready to initiate transcription. The positive function of cyclic AMP and CRP should be contrasted with the action of the lac and gal repressors whose presence prevents gene transcription. It is only when cyclic AMP and CRP are present and the repressor is neutralized by a specific inducer that transcription of the lac and gal genes can occur.
GENE ACTIVATION IN MAMMARY CELLS
Authors: Turkington Roger W. , Kadohama Nobuyuki
Published: October 2009

Hormonal activation of gene transcription has been studied in a model system, the mouse mammary gland in organ culture. Transcriptive activity is stimulated in mammary stem cells by insulin, and in mammary alveolar cells by prolactin and insulin. Studies on the template requirement for expression of the genes for milk proteins demonstrate that DNA methylation has an obligatory dependence upon DNA synthesis, but is otherwise independent from hormonal regulation of mammary cell differentiation. Incorporation of 5-bromo-2′deoxyuridine into DNA selectively inhibits expression of the genes for specific milk proteins. Undifferentiated mammary cells activate the synthesis of specific acidic nuclear proteins when stimulated by insulin. Several of these induced acidic nuclear proteins are undetectable in unstimulated undifferentiated cells, but appear to be characteristic components of the nuclei of differentiated cells. These results indicate that mammary cell differentiation is associated with a change in acidic nuclear proteins, and they provide evidence to support the concept that acidic nuclear proteins may be involved in the regulation of gene transcription and of mammary cell differentiation.
RIBONUCLEIC ACID SYNTHESIS AND OESTRADIOL ACTION IN THE UTERUS
Authors: Baulieu Etienne-Emile , Wira Charles R. , Milgrom Edwin , Raynaud- Jammet Claude
Published: October 2009

Some aspects of the fate of oestradiol in uterine cells and of a proposed »cascade« mechanism of its activity are discussed.

After a newly described step of entry of the hormone into the target cells, oestradiol is recognized by an intracellular transfer receptor system delivering it selectively to specific Non Histone Chromatin (NHC) protein of very feable abundance, provoking in turn (in a still unknown manner) the transcription of one (or a very few) gene(s). The resulting mRNA(s) would have a short half life and code for Key Intermediary Protein (KIP)(s) (also of very short half life). The latter activates the synthesis of essential component(s) of the cellular machinery in particular rRNAs, which are implicated post-transcriptionally in the generalized protein synthesis and therefore promote the growth of the differentiated cells. The reported changes in uterine RNA polymerase activity and protein synthesis are also discussed in the frame of other mechanisms.
ASSOCIATION BEHAVIOUR OF THE OESTRADIOL-BINDING PROTEIN COMPLEX WITH THE NUCLEAR FRACTION
Authors: Williams David L. , Gorski Jack
Published: October 2009

A number of studies have been carried out to examine the distribution of the oestradiol-binding protein complex between cytosol and nuclear fractions as a function of total binding site saturation. The results of these studies suggest that each binding protein has one binding site for the hormone. In addition, these studies suggest that the interaction of the oestradiol-binding protein complex with the nucleus involves a large number of low affinity association sites.
MORPHOLOGICAL ASPECTS OF PROTEIN SYNTHESIS
Authors: Droz Bernard , Pisam Monique , Chrétien Monique
Published: October 2009

Morphological aspects of protein synthesis have been mainly investigated by means of high resolution radioautography. Most of the cellular proteins are synthesized in the rough endoplasmic reticulum and on free cytoplasmic polysomes. Only few structural proteins (sedentary protein) remain in the vicinity of their sites of synthesis. The great majority of the newlysynthesized proteins move away (migratory protein). One portion of the migratory proteins is redistributed to various cell organelles and is referred to as "non exportable" protein. The migration of polypeptide chains from cytoplasmic polysomes ensures the renewal of protein in mitochondria, nucleus, and specialized membranes, in spite of the capacity of these organelles to incorporate slightly labelled amino acids. Plasma membrane precursors undergo a series of intracellular translocations and molecular complexification by adding carbohydrate groups before they reach the cell surface. Another portion of the migratory proteins, which includes secretory products, is referred to as "exportable" proteins. These exportable proteins are conveyed from the rough endoplasmic reticulum to the Golgi apparatus, then either stored and condensed in secretory granules or directly released by exocytosis. In some differentiated cells, the development of the cellular machinery for the synthesis of protein is triggered and maintained by the level of steroid hormones which controls both the quality and the quantity of secreted proteins.
EARLY STEPS IN THE FORMATION OF A TRANSLATION INITIATION COMPLEX ON NEWLY TRANSCRIBED MESSENGER RNA
Authors: Crépin Michel , Lelong Jean-Claude , Gros François
Published: October 2009

The aim of this study is to investigate how formation of a translation initiation complex affects the rate of transcription from a phage DNA template (λ plac or Ø 80 dlac DNA). Addition of "native" 30S ribosomes to a Ø 80 dlac in vitro transcription system (including limiting amounts of purified E. coli RNA polymerase and the requisite substrates) markedly enhances the rate of RNA synthesis. Factor free, 1 M NH4Cl washed 30S or 70S ribosomes show a weak albeit detectable activity, "run off 70S" or washed 50S being inefficient. Single addition of purified initiation factor IF3 greatly magnifies (about 2–3 fold) the stimulation obtained with washed 30S subunits, the effect being catalytic with respect to IF3; IF2 addition causes a weaker and stoichiometrical effect. Maximum stimulation (up to 6 fold) is achieved by the combined addition of washed ribosomes (30S or 70S), IF1, IF2, IF3 plus the highly purified fMet-tRNAfMet species. Under such conditions, very efficient initiator tRNA binding to nascent RNA does occur. Initiation factors show no activity in the absence of ribosomes. Kasugamycin greatly reduces the stimulation of RNA synthesis in the presence of the various translation elements.

This system provides a new and very sensitive means to study the various factor dependent ribosome-messenger interactions even in the absence of initiator tRNA, thereby enabling one to analyze early translation initiation steps.

Both the frequency of transcription initiation and the rate of RNA chain propagation appear to be enhanced when RNA synthesis and initiation of protein synthesis are coupled.
MATERNAL mRNA AND PROTEIN SYNTHESIS IN THE EMBRYO
Authors: Gross P. R. , Gross K. W. , Skoultchi A. I. , Ruderman J. V.
Published: October 2009

According to the masked maternal messenger RNA hypothesis, a large part if not all the protein synthesis of early development is directed by mRNA already present in the cytoplasm of unfertilized eggs. This mRNA is supposed to be synthesized during oogenesis and stored in some unavailable form until some later time in development, when it is selectively associated with the translational machinery. To the indirect evidence, which is nevertheless very strong, there can now be added a direct proof of the hypothesis for the case of histone mRNA. The five main histones of sea urchin embryos are synthesized on small polyribosomes, directed in part by newly-synthesized messages that sediment as a group at about 9S. Some histone synthesis survives total transcription block, however, suggesting that maternal histone mRNA exists. In competition-hybridization experiments, the egg RNA is shown to contain sequences characteristic of functional, embryonic histone mRNA. The competing RNA is localized in ribonucleoprotein particles of egg homogenates that sediment at 20–40S. These same particles contain RNA that stimulates a cell-free heterologous system to synthesize sea urchin histones. The application of these facts to some problems of translation control and of development generally is discussed.
SPECIFIC SECRETORY PROTEIN OF THE FEMALE GENITAL TRACT
Authors: Beier Henning M. , Beier-Hellwig Karin
Published: October 2009

Investigations on the biochemical composition of genital tract secretions during oestrus and early reproductive stages reveal a spectrum of different substrate and time specific patterns, particularly of secretory proteins. Results are presented on the biochemistry, immunological specificity, and endocrinological regulation of rabbit female genital tract secretion proteins. Special attention is drawn upon uteroglobin, which is one of the genital tract specific proteins presumably involved in preimplantational blastocyst development. Since analytical data on rabbit genital tract secretion proteins accumulate, there is consequently a necessity for an approach to comparable studies in the human uterus. Discelectrophoretical patterns and immunochemical test results are presented in this paper which encourage further studies on human genital tract specific proteins with regard to human reproduction, particularly with its contraceptive and fertility problems.
HUMAN CHORIONIC SOMATOMAMMOTROPIN IN DIABETIC PREGNANCIES
Authors: Wiese J. , Mølsted-Pedersen L. , Lebech P. E.
Published: October 2009

The serum concentration of human chorionic somatomammotropin (HCS) during the third trimester of pregnancy has been measured in 56 insulintreated diabetic women. A total of 318 serum samples were estimated and compared with a control material of 134 serum samples from normal pregnant women. The HCS concentration was found to be significantly higher in diabetic than in normal pregnancies. Significantly positive correlations were demonstrated between the HCS concentration and the placental weight, the birth weight of the infant and the oestriol excretion, respectively. Evidence is presented that simultaneous determinations of HCS and urinary oestriol would be valuable in the management of diabetic pregnancies.
OESTRIOL EXCRETION IN PREGNANT DIABETICS RELATED TO LONG-TERM PROGNOSIS OF SURVIVING CHILDREN
Authors: Yssing Minna
Published: October 2009

Urinary 24-hour oestriol excretion in pregnant diabetics were studied in 154 pregnancies resulting in 158 surviving children. Subnormal oestriol levels were found in 33 pregnancies associated with low birth weight, but otherwice widely independent on pre- and perinatal factors.

Results of follow-up study of the complete infant material at ages varying from 1 8/12 to 10 1/12 years were related to maternal oestriol excretion levels in relevant pregnancies.

The incidence of children with major abnormalities in the subnormal oestriol group (39.4 %) was significantly higher (P < 0.01) than in the normal oestriol group (16.0 %).

The incidence of cerebral damaged children in the subnormal oestriol group (51.5%) was significantly higher (P <0.01) than in the normal oestriol group (22.4 %).

There was no significant difference in distribution of congenital malformations.

Correlations of maternal White class, pre- and perinatal complications and birth weight with oestriol levels and results of follow-up study reflected the distribution in the total series with higher incidence of abnormal children in the subnormal oestriol fractions.
DESIGN OF EARLY CLINICAL TRIALS FOR NEW CONTRACEPTIVE METHODS
Authors: Mishell Daniel R. , Nash Harold A.
Published: October 2009

A plan is outlined to shorten the current suggested duration of animal testing prior to clinical investigation with certain types of pharmaceutical agents. For many new drugs, especially steroids, after a pharmacologic profile is established, long term toxicity testing can usually be shortened prior to initiation of 6 months clinical trials involving about 1000 women. Clinical trials of this magnitude usually correlate very well with ultimate performance of contraceptive drugs, provided proper selection of subjects is performed with resultant good continuation rates. It is thus possible with certain new antifertility candidates to safely shorten the duration of both toxicological and clinical studies before initiating large scale field trials.
SPECIAL TOXICOLOGY FOR THE PROSTAGLANDINS
Authors: Kirton K. T. , Carlson R. G.
Published: October 2009

The special requirements for toxicologic evaluation of the prostaglandins are considered. These compounds have many differences from the classical antifertility agents which seem to dictate special consideration for evaluation of safety and potential side effects. These include their ubiquitous distribution and spectra of pharmacologic activities, and extremely rapid in vivo metabolism to compounds of relatively low biological activity. All of these characteristics tend to cause prostaglandins to have a high order of acute activity, and therefore also acute toxicity. Most undesirable effects of steroids are seen only after prolonged administration. Toxicology and safety studies are designed to take these differences into account.
MACROPHAGE ACTIVATION INDUCED BY LYMPHOCYTE MEDIATORS
Authors: David John R.
Published: October 2009

This paper is a brief review of studies which demonstrate that lymphocyte mediators can activate macrophages in vitro. Macrophages which have been incubated in lymphocyte mediator-rich Sephadex fractions show changed morphology, metabolism and function. These changes include an increase in adherence to glass, ruffled membrane movement, phagocytosis of some particles, glucose oxidation through the hexose monophosphate shunt and an increase in the activity of a membrane enzyme, adenylate cyclase. Such mediator-activated macrophages show enhanced bacteriostasis and tumoricidal activity. In addition, studies describing the role of membrane sugars and esterases in the interaction of migration inhibition factor and macrophages are reviewed.
ONTOGENY OF IMMUNITY IN MAN
Authors: Stites Daniel P. , Caldwell Joseph , Carr Martin C. , Fudenberg H. Hugh
Published: October 2009

We have reviewed selected aspects of recent findings in the ontogeny of immunity in man. For obvious reasons, constraints placed on experimental work with human subjects, albeit deceased foetuses, limit the work to in vitro studies. Nevertheless, a number of novel and important concepts have emerged. First, as in lower animals, lymphoid development of the foetal thymus in general precedes the development of immunocompetence in peripheral lymphoid tissues. A striking exception to this rule is the finding of cells in early foetal liver which respond to allogeneic cells in the mixed lymphocyte culture some weeks before lymphoid organization of the thymus. In addition, the response of foetal cells to allogeneic cells in the mixed lymphocyte reaction (MLR) precedes the response to phytohaemagglutinin (PHA), a stimulant with relative T cell specificity. The use of markers for T cells to map the emergence of this class of lymphocytes correlates well with various functional attributes of T cells in in vitro culture systems. B cells emerge first in foetal liver at about 9 weeks' gestation, but formation of immunoglobulin by the foetus occurs at very low levels until after birth. There is a suggestion that progression from IgM to IgA synthesis occurs during human foetal lymphoid development. Taken as a whole, these data suggest a rather remarkable and perhaps unexpected degree of cellular and potential humoral immunocompetence at early stages of foetal development in man.
FERTILITY CONTROL THROUGH ACTIVE IMMUNIZATION USING PLACENTA PROTEINS
Authors: Stevens Vernon C.
Published: October 2009

Numerous studies have demonstrated that antibodies to placental proteins in a variety of species are capable of preventing or disrupting gestation. Early work in this area was primarily directed towards the passive immunization of rodents with heterologous antisera to whole placental extracts. Toxicity and renal damage often accompanied fertility inhibition. More recent studies reported less toxicity and a higher specificity of antibodies to reproductive function when anti-placental antibodies were absorbed with serum and extracts of non-reproductive organs. Few studies have been reported in which active immunization with placental proteins was employed. The most detailed studies of active immunization have employed highly purified placental hormones. Immunizations of rats and rabbits with human placental lactogen have resulted in marked reduction in reproductive function. Immunization of human females with chemically altered (hapten-coupled) HCG resulted in the production of antibodies reacting with unaltered HCG and pituitary LH. These antibodies were capable of reducing the level of endogenous serum LH in pre- and postmenopausal women. They also altered the events of the menstrual cycle in premenopausal women. More specific inhibition of chorionic gonadotrophin has been obtained by immunization of baboons with the beta subunit of HCG. Antifertility effects without alterations in the menstrual cycle of female baboons immunized with the beta subunit of HCG have been reported. The antibodies produced in these animals reacted significantly with human LH in vitro. The possibility of using hormonal and non-hormonal placental proteins as antigens for the specific immunological inhibition of fertility remains.