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Objectives: Isolated axillary lymph node metastases is an unusual clinical presentation of breast carcinoma. We studied its different issues., Methods: This study is a follow-up study of 16patients, treated between 1996 and 2012, presenting with axillary metastases with an occult breast carcinoma, which could not be identified by physical examination nor by a conventional imaging or a breast MRI. Clinical characteristics, histological analysis, treatment, monitoring and five-year survival rate were studied., Results: The incidence of this kind of breast cancer was 0.20%. A breast MRI was performed in 75% of the patients. The histology of these tumors showed a rate of hormono-sensibility of 50% and an HER2 overexpression of 44%. Sixty-nine percent of the patients had no breast surgery or radiotherapy; global five-year survival rate for these women was 77.4%±11.5., Conclusion: The survival rates of this study should lead the practitioner to choose a less aggressive breast therapy. Moreover, the histological characteristics explain the high metastatic potential of these tumors, and relate them to the HER2+ subclass of gene expression patterns of breast carcinomas., (Copyright © 2015. Published by Elsevier SAS.)

Biomarker assessment of breast cancer tumor samples is part of the routine workflow of pathology laboratories. International guidelines have recently been updated, with special regards to the pre-analytical steps that are critical for the quality of immunohistochemical and in situ hybridization procedures, whatever the biomarker analyzed. Fixation and specimen handling protocols must be standardized, validated and carefully tracked. Cooperation and training of the personnel involved in the specimen workflow (e.g. radiologists, surgeons, nurses, technicians and pathologists) are of paramount importance. The GEFPICS' update of the recommendations herein details and comments the different steps of the pre-analytical process. Application of these guidelines and participation to quality insurance programs are mandatory to ensure the correct evaluation of oncotheranostic biomarkers., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

The natural history of ductal carcinoma in situ (DCIS) is not fully elucidated, but it is recognized that DCIS is the true precursor of invasive carcinoma. Studies could show that DCIS is as heterogeneous as invasive ductal carcinoma, yet, they were unable to predict which DCIS will progress to invasion. Several biomarkers were also demonstrated to have a certain prognostic value. However, except for estrogen receptors and HER2, biomarkers are not yet widely used in clinical practice since their predictive value has not proven to be better than the grade and the classical classifying systems of DCIS. Identifying biomarkers for risk of invasiveness in DCIS could be of great value to help high risk patients through the management of their disease and to avoid overtreatment in low risk patients., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

The control of tooth development by 1,25-Dihydroxyvitamin D3 is analyzed by light- and electron-microscope immunocytochemistry and Northern-blotting in vitamin D-deficient rats. The receptor for 1,25-Dihydroxyvitamin D3, immunostained at the light microscope in all stem-cells, became immunodetectable only at the ultrastructural level in the ameloblasts which elaborate enamel and odontoblasts which synthetize dentin. Moreover, 1,25-Dihydroxyvitamin D3 induces an up-regulation specifically in these cells. In parallel, the calbindins-D9k, -D28k and osteocalcin, in contrast to the phosphoprotein, appear sensitive to vitamin D-deficiency. A single injection of 1,25-Dihydroxyvitamin D3 led to the increase of steady-state levels of the corresponding calbindin mRNAs. These data show that tooth constitutes a target-organ for 1,25-Dihydroxyvitamin D3, as other components of the phospho-calcic metabolism.

We have studied and compared the 1,25-dihydroxyvitamin D3 receptor (RD3) content of 154 human digestive carcinoma with the normal mucosa one, removed at distance from the same surgical specimen. The distribution of biopsies is as follows: 5 oesophagus, 10 stomach, 6 small bowel, 35 right colon, 47 left colon, 40 rectum and 11 pancreas. RD3 were measured by the Dextran Coated Charcoal method and characterized by sucrose gradient ultracentrifugation. One single class of high affinity binding sites (kD = 1.5 +/- 0.7 x 10(-10) M) was defined, with a sedimentation coefficient of approximately 3.4 S. The RD3 was present in the 6 samples of small bowel and in 82% of whole normal mucosa, whatever their localization along the digestive tract and pancreas, while in the tumoral tissue, the RD3 was positive in only 32% of the cases. In these tumor specimens the incidence decreases from 64% in the right colon to 27% in the left colon and only 15% in the rectum (P less than 0.001). RD3 rates vary slightly with the localization and are of the same level in normal tissues and in tumors: 10-314 (median = 59) vs 13-175 (median = 64) (P greater than 0.005) respectively. No significative variations relating to age or sex of patients were found. However, all RD3 positive tumors from left colon and rectum were well differentiated histologically. These results show that the normal colonic mucosa is a potential target for 1,25-dihydroxyvitamin D3, which can play a role in the metabolism of calcium and other ions. They also suggest that vitamin D3 could be a modulator of colorectal cell growth and differentiation and its receptor is frequently lost during malignant transformation.

The steroid receptor (ER, PgR) content of breast cancer specimens is now widely used as an indicator for hormone sensitivity which is a prerequisite for endocrine therapy. Is it conceivable to extend this steroid receptor determination to other types of tumors in order to select patients who would eventually benefit from an hormonal therapy? It seems that most normal or tumoral tissues contain detectable amount of steroid receptors. The values of each receptor comprise a continuum from very low to relatively high, suggesting that previous considerations of hormone dependence on the basis of presence or absence of hormone receptors may be oversimplified. The steroid receptor concentrations are however higher in target tissues, and could be of clinical interest in endometrium and prostatic cancers. The date are however insufficient to draw firm conclusions regarding their usefulness for treatment.

Benign breast diseases generally occur in women with progesterone insufficiency and an unopposed estrogen status. This hormonal imbalance is also a main risk factor for breast cancer in the long run. The presence of estradiol and progesterone receptors in benign breast disease are additional proof of their hormone dependence. The level of receptors is higher when the epithelial cellularity is high and also in more recent lesions, and younger women. The level and subcellular location of the receptors also depend on the endogenous secretion of steroids: their level varies throughout the menstrual cycle, and it is maximal at the end of the follicular phase, due to the high estradiol secretion. The cytosol and nuclear level and location of the receptors are also modified by hormonal therapy: estrogen-progestagens given as a contraceptive, or progestins given as a substitutive and antiestrogen therapy. These modifications constitute a biochemical basis for the hormonal progestagen treatment of benign breast diseases.

The degree of hormone-dependence of endometrial cancers may be evaluated by the presence and the rate of estradiol and mostly progesterone receptors in an endometrial fragment. The most differentiated tumors have a high receptor content, while anaplastic cancers are most of the time without them. They must be systematically determined regardless of other prognostic factors enabling to evaluate the prognosis and guide the treatment. It is presently facilitated by the method of immunochemical titration on histological section, available for estrogen receptors, disclosing the spatial and functional distribution of hormono-dependent cells.

There is less knowledge of androgen receptors than of oestrogen receptors but, in recent years, important progress has been made. The author thus describes the physico-chemical properties of the androgen receptors. He discusses also the problems raised by their estimation. The exchange techniques are practically unusable as the androgen receptors are easily destroyed even at 15 degrees C. The author proposes and describes another method which isolates the hormone-receptor complex with protamine sulphate. He also considers the prospects of these receptor estimations in prostatic disease, e.g. androgen-dependent adenoma. The absence of androgen receptors in testicular feminisation syndrome finally permits excellent illustration of the importance of the study of receptors in the understanding of cellular insensivity to a hormone.

In this preliminary study, we have determined the receptors in normal human prostatic tissue or from patients with benign prostatic hypertrophy and patients with prostatic cancer. A single point assay using a dextran-coated-charcoal treatment of cytosol is utilized. This method permits the elimination of endogenous steroids and is also useful when specimens are too small to provide the larger number of aliquots necessary for multiple steroid receptor analyses. The results are discussed in the light of several problems inherent to this assay. We conclude that adenoma-tissue contains more receptors than neoplasic tissues for the progesterones receptors but that there is no significant difference for the estrogen-receptor and the androgen-receptors. For all the adenoma tissues and some neoplasic tissues the level of the estrogen-receptors is always lower than the one for androgen-receptor which itself is lower than the one for progesterone-receptors. We think that the expression of the results as a ratio of two levels of steroid-receptors gives a better evaluation of the receptors. The ratios Progesterone-receptor Estrogen-receptor and Androgen-receptor/Estrogen-receptor are always greater than 1.5, except for some neoplasic tissues and we think that, as in breast tumors, there are perhaps two kinds of neoplastic tissues, one hormone-dependent, the other not.

Skin fibroblasts raised from Macaca fascicularis skin samples were tested for specific androgen receptors. By the 100th day of gestation male and female skin fibroblasts androgen receptors were detected. Higher levels of androgen receptors were found in sex skin fibroblasts. These preliminary results suggest that this primate could be used as a good model for the study of androgen receptors ontogenesis and regulation.

The development and normal and pathologic growth of the human prostate depends on testicular secretions. The human prostate contains receptors for the three sex steroids: testosterone, estradiol, and progesterone. The assumption that benign prostatic hyperplasia would result of a dihydrotestosterone excess is contradicted by the measurement of androgen receptors, which are not different in hyperplastic compared to normal prostate. Contrawise, hyperplastic prostate contains subnormal amounts of estradiol and progesterone receptors. The assay of hormone receptors does not seem to have any predictive value for the endocrine therapy of prostate cancer. However, tumors rich in nuclear androgen receptors or in dihydrotestosterone have longer remission interval and survival in response to hormone treatment.

In order to determine whether kidney hypertrophy secondary to unilateral nephrectomy was an appropriate model to study cytoplasmic androgen receptor modulation, this receptor was measured three weeks after surgery in intact or castrated animals. Our results demonstrate that the process of kidney hypertrophy is not accompanied by a proportional elevation of the number of androgen receptors and that these receptors are not significantly influenced by the gonadal activity in the rat.

This article reviews the problems inherent to the detection and assay of steroid hormone receptors in human target tissues and in pathological tissues. The biochemical characteristics of the hormone-receptor interaction are described, i.e., high affinity, strict specificity and limited number of binding sites. The calculation of these parameters is given. This is followed by an analysis of the practical problems involved in biochemical assays: (a) receptor protein stabilization, choice and concentration of radioligand, determination of incubation conditions (time and temperature) for the measurement of the number of binding sites in an exchange assay; (b) the various methods of separating bound from free steroid are discussed and their respective advantages and disadvantages as routine methods as opposed to research techniques are probed. The case of receptor determinations in human pathological tissues is analysed next with special emphasis on sample preparation, stabilization, storage and transport and also on plasma contamination and tissue heterogeneity. The various methods that can be used to detect steroid binding sites in pathological tissues are described: (i) biochemical techniques, (ii) autoradiography, (iii) histochemistry, in order to highlight their respective merits for the selection of hormone-sensitive pathological tissues.

In the course of our studies on the fate of cytoplasmic receptors for androgens in various target organs, such a receptor has been partially characterized in rat kidney cytosol: the high affinity constant, the specificity for testosterone and the formation of a hormone-receptor complex with a 8-10S sedimentation coefficient on sucrose density gradient led us to estimate that this macromolecule was similar to the rat prostate androgen receptor. This receptor is partially occupied by circulating androgens because the number of binding sites increases following castration or hypophysectomy. Constant levels of receptors have been recorded for several months in absence of testosterone. The age and the sex of animals, as well as the functional state of thyroid gland, did not affect the number of binding sites. These receptor characteristics may explain the slight renotrophic action of androgens in rat kidney.

This review deals with steroid hormones and receptors in relation to the physiology and the pathology of the central nervous system (CNS) and meninges. In recent years experiments performed in animals showed that: 1) endogenous steroid hormones cross the blood brain barrier: 2) radiolabelled steroid hormones bind in specific areas of the CNS; 3) all five classes of steroid receptors, i.e. oestrogen, progesterone, androgen, glucocorticoid and mineralocorticoid receptors (OR, PR, AR, GR, MR), are present in brain tissues, especially in the hypothalamus and the limbic system; 4) the interaction of steroid hormones and specific receptors induces the synthesis of proteins in the CNS; 5) finally, in situ metabolism of steroid hormones has been evidenced by the presence of specific enzymes. A few studies in human brain tissues have shown the presence of GR and OR as well as enzymes involved in the metabolism of sex steroid hormones. In neurology, some epidemiological and clinical data suggest the implication of steroid hormones and receptors in human CNS: 1) the influence of oestrogens in tardive dyskinesia; 2) the relevance of hormonal changes in benign intracranial hypertension; 3) the usefulness of glucocorticoid therapy in many patients with intracranial tumors and/or edema. Due to feasibility, most researches have concerned tumors: meningioma, neurinoma and glioma. Firstly, a reappraisal of biochemical and histochemical technics used to detect and characterize the receptors in tumors is presented. Then results from the recent literature are reviewed. In meningioma, PR was found in 89 p. 100 (152/177) of the cases, usually at moderate to high levels (up to 33 000 fmol/gT). In addition, PR has been fully characterized from a biochemical point of view. Furthermore, it has been hypothesized that PR may be a marker of leptomeningeal cells since it was detected at high levels in well differentiated tumors provided they had no or few psammoma. This was further supported by the discovery of PR in normal leptomeninges in human adults. OR was detected in 48 p. 100 (87/177) of the meningioma, at low levels. This is in contrast with PR but the percentage of cases with OR raises to 70 p. 100 (42/60) if one considers only tumors assayed for both cytosolic and nuclear receptors. Therefore it has been suggested that OR had translocated into the nucleus, at least in some cases, and subsequently the hypothesis of functional OR in meningioma was raised. AR was also detected in meningioma. Furthermore AR levels were found to correlate well with PR levels.(ABSTRACT TRUNCATED AT 400 WORDS)

Like other solid tumors ovarian tumors have been intensely investigated these past years to isolate specific tumoral antigens. These antigens are now used by clinical teams for post therapeutic follow-up. However, out of the 25 major antigens actually described in ovarian carcinomas, the only one that can be used today is the carcinoembryonic antigen (CEA). Moreover, an international collaborative work has demonstrated cross reacting antigenicity between these major tumoral antigens. Cellular enzymes (galactosyl transferase and sialyl-transferase) and hormonal receptors for proteinic hormones such as steroid hormones) are being studied but cannot be used as screening markers at this time.

The localization of mineralocorticoid and glucocorticoid binding sites along the nephron has been investigated by using an autoradiographic technique (dry film) and biochemical methods, applied on tubular segments of rabbit and rat kidney, isolated by microdissection, after in vitro incubation of kidney pyramids with tritiated aldosterone (Aldo) or dexamethasone (Dex). Based on analysis of dose-dependent specific nuclear binding values and competition studies, cortical collecting tubule (CCT) appears as the most specific mineralocorticoid segment; the distal tubule exhibits also high binding values, which are less specific for Aldo than in CCT; Aldo binding was lower in the ascending limb of Henle's loop and medullary collecting tubule and of mixed (gluco and mineralocorticoid) specificity. Aldo receptors were not present in proximal tubule and glomerulus. Specific nuclear binding of Dex was high all along the nephron, except in proximal tubules, and somewhat lower in glomeruli and its arterioles than in the distal parts of the nephron. Binding of 3H-Aldo did not differ drastically along the nephron of the rat and the rabbit.

Estrogen, progesterone and glucocorticoid receptors from 124 breast cancer cytosols were studied by high performance liquid chromatography (HPLC), in comparison with sephadex chromatography and density gradient ultracentrifugation. Sodium molybdate was used as a receptor-hormone complex stabilizer, and as a tool allowing to study the different molecular forms of the receptors. In the presence of molybdate during the entire saturation process, more receptor sites were detected, without modification of equilibrium reaction kinetics. HPLC receptors analysis was rapid, with an high relative separation efficiency. Multiple molecular forms of steroid receptors were observed: five forms each for progesterone and glucocorticoid receptors, and six forms for estrogen receptors. The presence or absence of one of these forms emphasized the biological variability of the tumors and provides further questions or hormone-dependence.

At the ultrastructural level, steroids and their receptors can be detected into their target cells by immunocytochemistry done on sections of anterior pituitary obtained by cryoultramicrotomy. An antigenic determinant is thus shown on fixed and frozen tissue with a monoclonal or polyclonal antibody. Tissue preparation is a compromise between preservation of ultrastructural features and the availability of the antigen. Specificity requires several controls knowledge of crossreaction of the anti-steroid or anti-receptor serum, immunological and biochemical analysis of the antigen, a check of the immunocytochemical reaction itself, specific absorbtion tests, modification to the immunocytochemical reaction in various experimental conditions. Specificity is also modulated by the dilution of the anti-steroid or anti-receptor serum as well as other experimental and physiological variations. Finally a hormone detected in its target cell may not be bound to its specific binding sites. Specific binding is characterized by: tissue and cell specificity, limited number or sites, hormonal specificity and a high affinity. Only thus is it possible to localise by immunocytochemistry the receptor or the hormone bound to its binding site.