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Biomedical subjects

E H Luque

Publications and source records attributed to E H Luque.

32 records · Page 2Linked to original sources
PubMed1997

Lack of relationship between the expression of Hsp27 heat shock estrogen receptor-associated protein and estrogen receptor or progesterone receptor status in male breast carcinoma.

Estrogen, through estrogen receptors (ERs), may regulate the synthesis of progesterone receptors (PRs) and of a heat shock estrogen receptor-associated protein (hsp27). In female breast carcinoma (FBC) both proteins serve as surrogate indicators for the presence of functional ERs. In addition, the expression of these proteins was related to other prognostic indicators of value in female breast tumours. Endocrine disorders, hormone therapy and altered estrogen metabolism have been associated with the development of male breast cancer (MBC), suggesting that evaluation of the expression of ER, PR and hsp27 might improve our understanding of the biology of this tumour. ER and PR status and hsp27 expression were evaluated by immunohistochemistry in 16 primary MBC patients. The interrelationships between these parameters were established and compared with the clinicopathological data on the tumours. Ten (56%) MBC patients were ER-positive, 69% were PR-positive and all samples were hsp27-positive. Our series of MBC patients showed a positive correlation between ERs and PRs, however there was a lack of correlation between hsp27 and ERs or PRs. MBCs did not exhibit any correlation between the biomarkers studied and known prognostic indicators for females (e.g. Scarff-Bloom-Richardson (SBR) or modified SBR (MSBR) grade, T stage, lymph node status). This is the first published series reporting the incidence of hsp27 in MBC. The lack of association between the expression of ERs and hsp27 found in MBC differs from the results reported for FBC, moreover the expression of ERs, PRs or hsp27 did not correlate with the clinicopathological parameters that have prognostic value in females. Although the data were obtained from a relatively small sample population, our findings suggest that MBC and FBC are biologically different tumours with respect to the expression of the studied proteins.

Adult↗
PubMed1997

Leukocyte infiltration and collagenolysis in cervical tissue from intrapartum sheep.

Sheep uterine cervices and cervical mucus were heavily infiltrated by neutrophils during labour, whereas samples of cervices obtained from non-pregnant controls had no infiltrate. The neutrophilic infiltrate of the sheep uterine cervix at term was not homogeneously distributed throughout the organ: luminal mucus contained more neutrophils than tissues which, in turn, displayed a differential distribution, the superficial subepithelial layer being more heavily infiltrated than the deeper submucous layers. A widespread collagenolysis was observed in the sheep uterine cervix at term. The homogeneous morphological aspect of degradation of collagen fibres throughout the whole cervical stroma contrasted with the above-mentioned differential distribution of neutrophils. On the basis of previous reports showing that collagenolysis follows the leukocytic invasion of human and rat cervices at term, a possible role for the neutrophilic infiltrate of the sheep uterine cervix is discussed.

Animals↗
PubMed1996

Dissociation in the control of cervical eosinophilic infiltration and collagenolysis at the end of pregnancy or after pseudopregnancy in ovariectomized steroid-treated rats.

Previous studies have demonstrated that during parturition the physiological ripening that permits dilation of the cervical canal is due to a widespread collagenolysis that follows a heavy polymorphonuclear leukocyte invasion of the uterine cervix. The aim of this study was to investigate whether there is any association between the ovarian steroid hormones involved in rat parturition and this phenomenon. Pregnant or pseudopregnant rats were ovariectomized (OVX) at Day 9 and then given a hormonal treatment (sufficient to maintain fetal viability) until Day 23. Cervical biopsies, taken from animals killed intrapartum or 1 h before expected parturition, were studied for eosinophilic infiltration and collagen birefringence. Intrapartum or sham-OVX pregnant rats showed a massive eosinophilic infiltration and a widespread collagenolysis as indicated by a loss of collagen birefringence. Ovariectomized pregnant rats treated with estrogen plus progesterone or with progesterone alone showed neither infiltration nor collagenolysis. In rats OVX during pseudopregnancy, estrogen given alone induced a significant infiltration of eosinophils in the cervical stroma; however, treatment with the combination of estrogen and progesterone was not able to promote eosinophilic infiltration. Collagenolysis was absent in all pseudopregnant animals. These results show that estrogen induced a cervical eosinophilic infiltration in rats but that when progesterone was added to the estrogen treatment the infiltration was not present; in addition, none of the steroid hormones assessed were responsible for the collagenolysis found in the cervical tissue at term.

Animals↗
PubMed1996

Correlation of thyrotropin secretion with morphometric data of secretory granules from individual thyrotropes. An ultrastructural study on rat cells identified by reverse hemolytic plaque assay.

We investigated the existence of functional subpopulations of thyrotropes in pituitary glands of male rats, also seeking to establish any correlation of thyrotropin secretion with morphometric data of secretory granules. Pituitary cells secreting thyrotropin were detected using a reverse hemolytic plaque assay. Population analysis of the plaque sizes (TSH secreted) of individual thyrotropes in control cultures from male rats revealed a unimodal frequency distribution. TRH added to the medium resulted in a shift to a bimodal distribution of sizes, consisting of small and large modes, but did not alter the fraction of plaque-forming thyrotropes. The plaque size of control cultures was the same as that of the fraction of TRH-stimulated thyrotropes forming small plaques. Because the extent of hemolysis is related to the amount of thyrotropin secreted by each individual thyrotrope, these results suggest that: a) all thyrotropes secrete thyrotropin even in the absence of a stimulatory dose of TRH; b) the thyrotrope subpopulation secreting small amounts of thyrotropin when incubated with TRH may be unresponsive to this stimulatory factor. After performing reverse hemolytic plaque assay, plaque-forming cells were subsequently processed for electron microscopy. Morphometric analysis of the secretory granules correlated well with plaque size (TSH secreted). It is thus concluded that the functional subtypes of thyrotropes can be recognized on the basis of their ultrastructural images.

Animals↗
PubMed1993

[Molecular markers to predict the prognosis of breast cancer].

The incidence of breast cancer is 25-30% of all malignant female tumors and represents the highest rate of mortality. To define those breast cancer patients at higher risk several prognostic factors are routinally evaluated in the primary tumor. Among them, the presence and degree of tumor involvement in axillary lymph node is one of the most powerful prognostic indicator. However, around 30% of node negative patients (good prognosis group) have recurrences and die within the next 10 years from diagnosis. Therefore, there is a need for markers to better discriminate biologic differences in the primary tumors. The techniques of molecular biology are shedding new insight into the subcellular pathology of malignancy. The crucial events of carcinogenesis, tumor progression, and metastatic spread are coming into focus at a molecular level. In some tumors, these advances in the laboratory are beginning to have applications at the bedside. Recently, abnormalities in the copy number and expression of several genes have been correlated with prognosis of individual patients with selected types of cancer. Molecular biology laboratories can provide useful predictive information that can be used to influence decision on the selection of treatment and to estimate a better risk stratification. Among these new markers are: oncongene amplification and/or over-expression, growth factors, cellular proliferation rate and ploidy, estrogens induced proteins, expression of metastasis related molecules, drug resistance associated proteins, etc. In this article we attempt to present a brief evaluation of a new technology that promises to add greater precision in evaluating molecular tumor markers and we mention some of its clinical applications.(ABSTRACT TRUNCATED AT 250 WORDS)

Biomarkers, Tumor↗
PubMed1991

Immunological evidence for the identity between the hsp27 estrogen-regulated heat shock protein and the p29 estrogen receptor-associated protein in breast and endometrial cancer.

Previous studies have shown several similarities between an estrogen-regulated heat shock protein of 24,000-28,000 daltons (hsp27), and a cytoplasmic estrogen receptor-associated protein of 27,000-29,000 daltons (p29). These proteins have been studied by monoclonal antibodies generated in different laboratories. In the present report we have performed immunocytochemical and immunoblot studies to explore if the monoclonal antibodies against hsp27 (C11) and against p29 (ER-D5) may be identifying the same protein. Breast and endometrial carcinomas and normal endometrial samples were examined by immunocytochemistry (in mirror sections and by double-immunostaining). Identical hsp27 and p29 immunostaining intensity, distribution, and percentage of stained cells was demonstrated by immunocytochemistry. The antigens examined by the two antibodies appeared in the same cells. Cytosols from tumors analyzed by the Western blot technique revealed that the C11 and the ER-D5 antibodies recognized bands with identical electrophoretic mobility. Immunoprecipitation studies with one antibody, C11, followed by Western blot showed that the precipitate was reactive with both antibodies. Identical C11 and ER-D5 reacting spots were observed after blotting proteins separated by high resolution two-dimensional gel electrophoresis. In addition, p29 protein was induced by heat shock in the estrogen receptor negative MDA-MB-231 human breast tumor cell line. These results strongly suggest that the two proteins under investigation are identical.

Antibodies, Monoclonal↗
PubMed1989

Progesterone promotes a massive infiltration of the rat uterine cervix by the eosinophilic polymorphonuclear leukocytes.

Rat uterine cervices contained an eosinophilic infiltrate during labor, whereas samples of cervices obtained from nonpregnant controls had no infiltrate. A heavy eosinophilic invasion, which closely resembled that of the cervix uteri at term, could be reproduced in ovariectomized rats following progesterone administration, but not in spayed rats injected with estrogen. Light and electron microscopic evidence showing a widespread collagenolysis in the cervical stroma at term is presented. This collagenolysis follows the eosinophilic invasion. On the basis of these observations, a possible role for the eosinophilic infiltrate of the uterine cervix is discussed.

Animals↗
PubMed1988

Effects of ovarian steroids on vaginal smears in the rat.

A correspondence between the appearance of vaginal smears and the layers of the epithelium from which the cells had desquamated was established in untreated rats during the estrous cycle, in control ovariectomized rats and in spayed rats injected with either estrogen or progesterone. The technique for preparing and staining the smears (modified Shorr's staining procedure) is outlined. A simplified system of classification which allows the accurate identification of the various stages of the reproductive state in the rat is described. Standing estrus, as well as the influence of estrogen on spayed rats, is characterized by marked cornification of the cells and the disappearance of leukocytes. At the end of estrus, the cornified layer is sloughed off and invasion by leukocytes occurs. During diestrus, as well as in untreated ovariectomized rats, the vaginal contents consistently lack cornified cells whereas leukocytes are very plentiful. Proestrus follows diestrus: the vaginal smear is devoid of leukocytes and characterized by nucleated epithelial cells. Pregnancy, as well as the influence of progesterone on ovariectomized rats, is also characterized by epithelial growth and desquamation but at different rates, resulting in the presence of intermediate cells, and polymorphs and mucus forming a noticeable background to the smear. Since vaginal smears display cell pictures characteristic for each hormone after administration of estrogen or progesterone, exfoliate cytology is a good indicator of the stage of the reproductive state in the rat.

Animals↗
PubMed1986

Detection and measurement of secretion from individual neuroendocrine cells using a reverse hemolytic plaque assay.

Recent advances in technology have dramatically increased the resolution with which we may examine many features of biological systems. Intracellular recording and tracer injection techniques allow one to study the function of individual neurons and later characterize the same cells morphologically. In situ hybridization techniques can give us information about messenger RNA levels in single cells. More established techniques such as immunocytochemistry and electron microscopy also provide information at the cellular and even subcellular level. With each of these technological advances we have learned more about the mechanisms underlying cell function. We are also beginning to appreciate the role of heterogeneity among cells in relation to the function of the whole organism. Application of the reverse hemolytic plaque assay to the study of hormone or neurotransmitter secretion should help clarify this role. This technique permits accurate quantitation of hormone secreted from a large number of cells. Thus while cells can be studied individually they can also be categorized into functional subpopulations. As discussed in this chapter, many other techniques may be applied on cells which have already been functionally defined with the plaque assay. This should result in a clearer understanding of the roles of secretagogue binding and internalization, activation of second messenger systems, protein synthesis, and the cytoskeleton in hormone secretion. In the plaque assays described in this chapter individual pituitary cells are isolated in culture free from possible interactive effects coming from other cells. While these interactions are no doubt critical to the understanding of the function of the organism as a whole they can result in totally uninterpretable results. In fact, when we have gained some understanding into the functioning of individual cells it should be possible using the plaque assay to study the interactions among cells in a controlled fashion.

Adrenocorticotropic Hormone↗
PubMed1986

Subpopulations of lactotropes detected with the reverse hemolytic plaque assay show differential responsiveness to dopamine.

Cultured adenohypophysial cells secreting PRL were detected with a reverse hemolytic plaque assay. In this assay, PRL secretion from a pituitary cell results in hemolysis of cocultured protein A-coupled ovine erythrocytes in the presence of PRL antiserum and complement, so that a zone of hemolysis (a plaque) surrounds each lactotrope. The extent of hemolysis was related to the amount of PRL secreted by each lactotrope: batches of cohort cells incubated under similar conditions either in petri dishes for measurement of PRL secretion by RIA or in Cunningham chambers for measurement of plaque area revealed a significant relationship between secreted PRL and plaque area (r = 0.97; regression coefficient = 0.0007 pg/micron2). Measurement of plaque area on lactotropes derived from proestrous rats revealed a bimodal frequency distribution that was composed of cells forming small plaques (35% of the total lactotrope population) and others forming large plaques (65%). Treatment with 10(-7) M dopamine appeared to preferentially inhibit the large plaques; they decreased to 42% of the total with corresponding increases in the number of small plaques, but the total number of secretory lactotropes did not change. At 10(-5) M dopamine, large plaques virtually disappeared (only 9% remained), and small plaques appeared in increased numbers, but the number of secretory lactotropes decreased by about one third. These results suggest that the reverse hemolytic plaque assay can be used to quantify PRL secretion by individual lactotropes, that lactotropes from proestrous rats exist as two secretory subpopulations, and that dopamine may preferentially suppress the subpopulation secreting large amounts of PRL.

Animals↗
PubMed1984

Modulation of sensitivity to cervicovaginal stimulation during the estrous cycle: evidence for an extrapituitary action of LH-RH.

Single stimulations of the vagina and cervix were performed between proestrus and the first day of diestrus with a stimulator designed to grade the intravaginal penetration of a rod. The percent incidence of pseudopregnancy after this stimulation was exponentially related to the extent of intravaginal penetration and was also affected by the stage of the cycle at which the stimulation was performed. At 10.00 h on proestrus, an exponential increase in the incidence of pseudopregnancy was observed with shallow penetrations, while an exponential decrease was found when deeper penetrations were applied. Such negative exponential correlation had disappeared at 22.00 h on proestrus. At that time, also, some responses were elicited by very shallow penetrations (17 mm) and all the animals responded to penetrations of 20 mm or more. Sensitivity to cervicovaginal stimulation at 10.00 h on estrus was lower than that at 22.00 h on proestrus and it was even lower at 10.00 h on the first day of diestrus. The response to 18 mm of penetration was studied every 3 h between 10.00 h on proestrus and 10.00 h on estrus, and then every 12 h until 10.00 h on the first day of diestrus. This stimulation was usually ineffective to induce pseudopregnancy, except for a brief period encompassing the night between proestrus and estrus, when a peak in the incidence of responses was reached. This peak sensitivity could be advanced following the s.c. administration of 250 and 500 ng of LH-RH at 11.00 h on proestrus. Other doses were ineffective. The peptide (500 ng) was unable to induce pseudopregnancy in rats that received no cervicovaginal stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Sensory mechanisms involved in the induction of pseudopregnancy by progesterone: increased sensitivity to stimulation of the pudendal sensory field.

The sensory mechanisms that participate in the induction of pseudopregnancy after a single injection of progesterone were investigated. Unless otherwise indicated, rats were kept in group cages and vaginal smears were taken daily. Progesterone evoked pseudopregnancy in a dose-dependent manner when administered to proestrous or estrous rats that received no cervicovaginal stimulation. The probability of pseudopregnancy after progesterone was higher on estrus. Cervicovaginal stimulation of proestrous rats that received 5 mg progesterone 10 h before was performed with a rod with a sliding stop attached to regulate its intravaginal penetration. Progesterone facilitated responsiveness to this stimulus, although the amount injected was not significantly effective in increasing the incidence of pseudopregnancy in nonstimulated rats. However, the mere application of the stop of the stimulator on the perineal skin was followed by a significantly higher incidence of pseudopregnancy in progesterone-injected rats than in their vehicle-injected controls, which suggested an action of the steroid on perineal sensitivity. Accordingly, the pseudopregnancy-evoking effect of progesterone was clearly inhibited by refraining from taking vaginal smears for 5 days after steroid injection on estrus. No further inhibition was observed after isolating the animals in single rat cages. However, daily finger stimulation of the perineal skin of nonsmeared rats restored to a normal level response to progesterone. Furthermore, this response was severely impaired by transecting the pudendal nerves before the injection. It is concluded that pseudopregnancy is induced in progesterone-treated rats through sensory stimulation of the pudendal receptive field and it is suggested that the pudendal nerve may subserve as a secondary afferent system to elicit the pseudopregnancy response. The possibility progesterone also acts on other afferent systems including the main afferent system constituted by the pelvic nerve is discussed.

Animals↗
PubMed1983

A histochemical and electron microscopic study on the collagen of nerves in the domestic fowl.

The collagen in the endoneurium is present as argyrophilic reticular fibres, which show up as thin, weakly birefringent greenish fibres when studied by aid of the Picrosirius-polarization method, and are composed of loosely arranged thin collagen fibrils. The epineurium consists of thick non-argyrophilic collagen fibres, which display a strong birefringence of red or yellow colour when studied by the aid of the Picrosirius-polarization method, and consist of closely packed thick collagen fibrils. These characteristics strongly suggest that the reticular fibres in the endo- and perineurium are composed mainly of collagen Type III, whereas collagen Type I predominates in the epineurium. The fact that these observations on birds are consistent with the descriptions available in the literature for fish, amphibians, reptiles and mammals, argues in favour of the existence of a uniform structural pattern of collagen distribution that is a general phenomenon in vertebrate nerves.

Animals↗