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M F Press

Publications and source records attributed to M F Press.

At least 55 records · Page 3Linked to original sources
PubMed1990

HER-2/neu oncogene expression and DNA ploidy analysis in breast cancer.

This study was performed to evaluate the correlation between HER-2/neu gene expression and DNA ploidy patterns. Forty-five cases of breast-cancer were analyzed. Immunohistochemical staining of HER-2/neu protein on frozen sections was used to detect the HER-2/neu protein, and the Feulgen DNA staining method was used to assess DNA amounts in the same tumor cells. Positive HER-2/neu overexpression was evaluated visually, and quantitation of the HER-2/neu protein was measured by image analysis. Twenty-two of the 45 cases were visually scored to be positive for the overexpression of the HER-2/neu protein, and these cases also contained above 10% HER-2/neu protein compared with a standard control cell line. All 22 of these cases had near-tetraploid DNA content. In contrast, cells, derived from the 23 cases that did not overexpress the HER-2/neu protein, contained DNA amounts that ranged from euploid (diploid) to varying degrees of aneuploid. The results of this study indicated that tumors that overexpress the HER-2/neu protein have tetraploid or near-tetraploid DNA content. This pattern could relate to the biological behavior of these tumors.

Adenocarcinoma↗
PubMed1990

Expression of the HER-2/neu proto-oncogene in normal human adult and fetal tissues.

The HER-2/neu proto-oncogene is homologous with, but distinct from, the epidermal growth factor receptor. Current evidence indicates that this gene is frequently amplified and/or overexpressed in some human breast and ovarian cancers and that these alterations may be clinically important; however, little is known about the expression pattern of the gene in normal tissues. Using immunohistochemistry and northern blot analyses to identify the HER-2/neu protein and transcript respectively, we have evaluated a variety of normal adult and fetal tissues for HER-2/neu expression. HER-2/neu protein was identified on cell membranes of epithelial cells in the gastro-intestinal, respiratory, reproductive, and urinary tract as well as in the skin, breast and placenta. Northern hybridization confirmed the presence of the 4.5 kb transcript encoding the protein in these tissues. The amount of HER-2/neu message and protein was generally higher in fetal tissues than in the corresponding normal adult tissues. HER-2/neu expression levels in these normal tissues were similar to the levels found in non-amplified, non-overexpressing breast cancers and breast cancer cell lines. Southern hybridization of extracted DNA showed that none of the normal tissues expressing HER-2/neu had amplification of the gene. These results confirm that HER-2/neu is normally a membrane constituent of a variety of epithelial cell types.

Blotting, Northern↗
PubMed1989

Comparison of the quantity of estrogen receptors in human endometrium and myometrium by steroid-binding assay and enzyme immunoassay based on monoclonal antibodies to human estrophilin.

The recently developed enzyme immunoassay for estrogen receptors is more simple to perform with quality assurance than conventional steroid-binding assays with radioactive labeled estrogen. However, it is not known how well the results of the two assays agree for normal human uterine samples. We compared enzyme immunoassay (Abbott estrogen receptor enzyme immunoassay) and steroid-binding assay of normal human endometrium and myometrium. Low-salt, "cytosol" estrogen receptor determinations were performed by dextran-coated charcoal assay, and high-salt, "nuclear" estrogen receptors were measured by controlled pore glass bead assay. Results showed excellent correlation (p less than 0.0001) for cytosol estrogen receptor of endometrium (r = 0.95) and myometrium (r = 0.79) and also for a smaller number of nuclear estrogen receptors of myometrium (p less than 0.01, r = 0.69). Good agreement between steroid-binding assay and enzyme immunoassay was seen for estrogen receptors of both proliferative and secretory phase samples. Thus the data indicate that the simpler estrogen receptor enzyme immunoassay is useful to measure estrogen receptor in the normal uterus. Furthermore, with this sandwich assay, there is no evidence for the existence of significant quantities of receptor fragments that do not bind estrogen.

Adult↗
PubMed1989

The effect of the alpha-emitting radionuclide lead-212 on human ovarian carcinoma: a potential new form of therapy.

To improve response and survival of patients with ovarian carcinoma noncross-resistant forms of therapy must be developed. alpha-emitting radionuclides may be therapeutically useful since they can directly ionize with energies of 5 to 9 MeV, penetrate only a few cell diameters, and transfer a high amount of energy. The purpose of this study was to determine the effect of the alpha-emitter, lead-212 (212Pb), complexed to sulfur in a nude athymic mouse model (NIH:OVCAR-3) containing human ascites and solid epithelial ovarian carcinoma. Thirty-six nude mice 28 to 32 days old were injected with 10(7) to 10(8) carcinoma cells from donor mice. After 4 weeks, six groups of six nu/nu athymic BALB-C mice were intraperitoneally injected with 70, 50, 20, 5 microCi of 212Pb sulfur colloid, sulfur colloid, or saline. Tumor necrosis with a decrease in ascites and a dose-related survival were noted with doses of 50, 20, and 5 microCi. With 70 microCi acute gastrointestinal toxicity developed. These experiments form the basis for further investigations and the development of alpha-emitting radiocolloids which may be of therapeutic efficacy in the treatment of intraperitoneal ovarian carcinoma.

Animals↗
PubMed1989

Estrogen and progesterone receptors in meningiomas: comparison of nuclear binding, dextran-coated charcoal, and immunoperoxidase staining assays.

We studied the status of estrogen (ER) and progesterone (PR) receptors in meningiomas removed from 52 patients, comparing dextran-coated charcoal (DCC), nuclear binding (NB), and immunoperoxidase (IP) assays. Each of the assays was performed independently by investigators well-experienced with these assays. The NB assay is a new assay that measures functional steroid receptors--that is, the activation of the receptor and its binding to the nucleus. The assay is very sensitive and requires a relatively small amount of tissue as compared with the DCC assay. In agreement with data from other studies. PR were detected in most meningiomas by all 3 methods: in 69% of the cases by NB, in 76% by DCC, and in 89% by IP. ER were detected in only a few cases: in 33% by NB, in 2% by DCC, and in none by the IP assay. The agreement for PR sites was 62% for all 3 assays; it was 66% between the NB and DCC assays, 67% between the NB and IP assays, and 86% between the DCC and IP assays. Of 26 cases that were positive by the DCC assay, 6 (23%) were negative by NB. The overall agreement for all three ER assays was 65%. The data suggest that the majority of meningiomas contain high-affinity receptors for progesterone, that estrogen receptors are present in only a few meningiomas, and that some of these estrogen and progesterone receptors appear to be functional.

Adolescent↗
PubMed1989

Subcellular distribution of estrogen receptor and progesterone receptor with and without specific ligand.

The estrogen receptor (ER) and progesterone receptor (PR) content of cultured human breast carcinoma cells (MCF-7) was determined by biochemical assay, immunoblot analysis, and immunohistochemical assay under varying conditions of hormonal stimulation. The ER and PR content in cytosolic and nuclear extracts varied with steroid treatment. However, both the amount and distribution of each receptor in these extracts was virtually the same when determined by steroid binding and immunoblot analyses. Two immunocytochemical parameters (staining intensity and proportion of cells stained) correlated with the quantitative analyses of ER and PR, but not with the subcellular distribution. When MCF-7 cells were grown for 4 days in charcoal-stripped serum without phenol red, 93% of total ER was found in the cytosol (10 mM KCl), whereas short-term treatment with 5 nM estradiol resulted in the appearance of 82% of total ER in the nuclear extract (400 mM KCl). With either cell treatment only nuclear staining for ER was observed. Progesterone receptor was virtually undetectable in the same cells by any method. After 4 days of treatment by 5 nM estradiol, PR was strongly induced (50-fold) in MCF-7 cells as determined by all three methods. As observed for ER, 95% of total induced PR was found in the cytosol in the absence of a progestin. Short-term treatment with 5 nM ORG 2058, a synthetic progestin, resulted in the appearance of 42% of total PR in the nuclear extract. However, only strong nuclear staining for PR was observed in either the presence or absence of a progestin. These findings are consistent with the current view of ER and PR as nuclear receptors present in at least two forms. One of these, the unoccupied form of the receptor, is easily removed from the nucleus by hypotonic buffers during the cell homogenization process and appears in the cytosolic extract. The other form of the receptor, the steroid-occupied form, is more tightly bound to nuclear components and is removed from nuclei only under more vigorous extraction conditions.

Blotting, Western↗
PubMed1988

The evaluation of estrogen receptor in primary breast carcinoma by computer-assisted image analysis.

A monoclonal antibody prepared against estrogen receptor has been shown to be specific and sensitive for the detection of estrogen receptor in human breast lesions by use of immunohistochemical methods. Two hundred selected cases of primary breast carcinoma were assayed for estrogen receptor content by biochemical and immunohistochemical procedures. Quantitative evaluation was by biochemical, immunohistochemical, and automated computer-assisted image analysis using the Cell Analysis System's CAS/100 machine (Lombard, IL). Quantitative estrogen receptor content was determined by dextran-coated charcoal analysis and sucrose density gradient analysis. Immunohistochemical evaluation incorporated both intensity and distribution of staining, yielding a subjective score, histologic score (HSCORE). An objective quantitation, also incorporating intensity and distribution of staining, was done by computer-assisted image analysis, quantitative immunocytochemical score (QIC SCORE). HSCORE analysis was done with and without methyl green counterstain with no loss of sensitivity. Comparison of QIC SCORE with the biochemical and immunohistochemical analysis of the tissues examined revealed excellent sensitivities and specificities. These data suggest that automated image analysis provides an effective qualitative and quantitative means of evaluating estrogen receptor content in human breast cancers.

Antibodies, Monoclonal↗
PubMed1988

Subnuclear localization of the trans-activating protein of human T-cell leukemia virus type I.

Human T-cell leukemia virus type I is associated with human lymphoid malignancies. The p40xI protein encoded by the x gene of this virus is believed to play some role in virally mediated transformation. This gene is known to encode a transcriptional trans activator which previous studies have shown to be a nuclear protein. Further characterization of the intracellular kinetics of this protein showed that it migrated into the nucleus very soon after synthesis. Within the nucleus, p40xI was distributed almost equally between the nucleoplasm and the nuclear matrix. Given the proposed role of the nuclear matrix in RNA transcription, the association of p40xI with the matrix places it in an appropriate cellular compartment to exercise an effect on transcription.

Cell Compartmentation↗
PubMed1988

Localization of progesterone receptor with monoclonal antibodies to the human progestin receptor.

A series of rat and mouse monoclonal antibodies to the human progesterone receptor (PR) has recently been produced. These antibodies were used for the immunocytochemical identification of PR in several mammalian species including humans. The specificity of the monoclonal antibodies for PR was confirmed by using competition studies with purified PR and by comparison of the immunostained tissues, known from steroid binding assays to be receptor rich, with immunostained tissues known to be receptor-poor. Immunoreactive PR was found in the nuclei of uterine epithelial, stromal, and smooth muscle cells; benign ductal and lobular epithelial cells of the breast; ovarian surface epithelium; ovarian stroma and luteal cells; pulmonary parenchymal cells; and selected pituitary parenchymal cells. A proportion of the following selected human tumors expressed PR: breast carcinomas, endometrial carcinomas, ovarian carcinomas, and meningiomas. PR was localized to the nuclei of all progesterone target tissues even under conditions where the vast majority of the receptor is unoccupied by steroid, suggesting that the unoccupied as well as the steroid-occupied form of PR are predominantly nuclear proteins, as observed previously for estrogen receptor and rabbit PR.

Adult↗
PubMed1988

Progesterone receptor distribution in the human endometrium. Analysis using monoclonal antibodies to the human progesterone receptor.

Two monoclonal antibodies to the human progesterone receptor (PR), JZB39 and KD68, were used in determining the immunohistochemical distribution of PR in the human endometrium throughout the menstrual cycle and after menopause. These antibodies recognized PR, as demonstrated by a downfield shift in the radiolabeled progestin binding peak when KD68 or JZB39 was added to high salt sucrose density gradients. The specificity of both antibodies for PR was confirmed with Western immunoblots and competition studies performed with purified receptor. Progesterone receptor was identified with these antibodies and the peroxidase-antiperoxidase technique in the nuclei of epithelial cells, stromal cells, and myometrial smooth muscle cells. The receptor content of endometrial epithelium and stroma varied with the menstrual cycle. The variation was most marked in the epithelium, which demonstrated very strong PR immunostaining during the proliferative phase and postovulation Days 1-3 of the early secretory phase, but PR immunostaining decreased sharply at postovulation Day 4 and remained relatively weak or absent during the mid and late secretory phase. In contrast, stromal cell nuclei were moderately to strongly immunostained even during the secretory phase. Progesterone receptor was not localized in vascular smooth muscle cells or endothelial cells. Specific cytoplasmic staining for PR was not identified in any of these cases, even prior to ovulation, when circulating levels of progesterone are low, indicating that both the steroid-occupied and -unoccupied forms of human progesterone receptor, like rabbit and guinea pig PR, and estrogen receptor, is a nuclear protein.

Antibodies, Monoclonal↗
PubMed1987

Estrogen receptor localization in formalin-fixed, paraffin-embedded endometrium and endometriotic tissues.

Monoclonal estrogen receptor (ER) antibody and indirect immunoperoxidase techniques were used to study the immunohistochemical localization of ER in paraffin sections of 20 surgical specimens containing foci of endometriosis. The eutopic endometrium, available in 16 of 20 cases, was dated histologically and stained for ER. Specific nuclear staining was observed in glandular and stromal cells of the endometrium during the proliferative and early secretory phases. Mid and late secretory phase endometria showed only focal staining. Dating of the endometriotic tissues using morphologic criteria revealed that these were in phase with the endometrium in nine of 16 cases. In these cases the ER staining of both the endometriotic tissues and endometrium was similar. The endometriotic tissues could not be dated as either proliferative or secretory in seven cases and stained variably for ER. Diffuse nuclear staining for ER was present in the endometriotic glands of three of these cases in which the endometriotic glands morphologically resembled glands of the basalis contrasting with weak, focal staining in the corresponding eutopic secretory endometria. The majority of endometriotic tissues examined mimicked the cyclic ER expression of the eutopic endometrium and thus appeared to correspond to the morphologic appearance of the endometriotic glands. Immunohistochemical staining with monoclonal estrophilin antibodies may be helpful in supporting the histopathologic diagnosis of endometriosis.

Adult↗
PubMed1986

Studies of the human c-myb gene and its product in human acute leukemias.

The myb gene is the transforming oncogene of the avian myeloblastosis virus (AMV); its normal cellular homolog, c-myb, is conserved across a broad span of evolution. In humans, c-myb is expressed in malignant hematopoietic cell lines and in primary hematopoietic tumors. Partial complementary DNA clones were generated from blast cells of patients with acute myelogenous leukemia. The sequences of the clones were compared to the c-myb of other species, as well as the v-myb of AMV. In addition, the carboxyl terminal region of human c-myb was placed in an expression vector to obtain protein for the generation of antiserum, which was used to identify the human c-myb gene product. Like v-myb, this protein was found within the nucleus of leukemic cells where it was associated with the nuclear matrix. These studies provide further evidence that c-myb might be involved in human leukemia.

Aspartate Carbamoyltransferase↗
PubMed1986

Structure and dynamics of the estrogen receptor.

To evaluate the structure and function of estrogen receptor (ER) in various mammalian systems, the cytosolic forms of receptor from calf uterus and from MCF-7 human breast cancer cells have been purified to virtual homogeneity by sequential selective adsorption to estradiol-Sepharose and heparin-Sepharose. In both cases, the purified steroid-receptor complex appears to exist as an activated 5S homo- or heterodimer of mol. wt 65,000 (4S) steroid-binding subunits. Purified ER has high affinity for DNA and serves as a substrate for phosphorylation by a purified rat brain kinase. Several monoclonal antibodies prepared against affinity-purified MCF-7 cytosol ER have been used to localize receptor by an indirect immunoperoxidase technique in fixed, frozen sections of human breast tumors, human uterus, rabbit uterus and in other mammalian reproductive tissues and cancers, as well as in fixed MCF-7 cell cultures and in paraffin-embedded sections of breast tumors and human endometrium. In all cases, we have observed only nuclear localization of immunoreactive receptor in tissues and whole cells, even under conditions in which virtually all of the receptor is found in a low-salt extract (cytosol) of the target cells. Treatment of cells or tissues in vivo or in vitro with estradiol alters the intensity but not the distribution of specific staining for ER. By immunoelectron microscopy, receptor was localized in the euchromatin, but not in the marginated heterochromatin or nucleoli of MCF-7 nuclei and epithelial and stromal nuclei of postmenopausal human endometrium. These observations suggest that the majority of the unoccupied receptor may actually reside in the nucleus, rather than in the cytoplasm as previously thought. Thus, hormone action may involve binding of the steroid directly to receptor loosely associated with nuclear components, followed by conversion of the steroid-receptor complex to an activated form which becomes more tightly associated with chromatin.

Animals↗
PubMed1986

Golgi-electron microscopic study of sprouting endothelial cells in the neonatal rat cerebellar cortex.

Intraparenchymal postnatal development of the CNS vasculature proceeds by a process of vascular 'budding' or 'sprouting' from existing endothelial cells and elongation of these immature vessels. A modified Golgi technique combined with gold toning and deimpregnation was used to identify and characterize sprouting endothelial cells in the neonatal rat cerebellar cortex with light and electron microscopy. Sprouting endothelial cells were the terminal endothelial cells of immature, developing blood vessels and they had characteristic morphologic features. The most distinctive feature was an array of tentacular processes (0.1-0.2 micron in diameter and ca. 20 microns in length) which radiated from the apex of these cells in the presumed direction of vessel growth. Cytoskeletal microtubules and microfilaments were the characteristic organelles of these tentacles. Sprouting endothelial cells were thin (1-3 microns in diameter). lacked a vessel lumen and basement membrane, had abundant cytoplasmic organelles and a prominent nucleus and were closely associated with the subterminal endothelial cell without interendothelial gaps. The developing blood vessels had a blood-brain barrier which excluded intravenously injected colloidal carbon from the neuropil.

Animals↗
PubMed1986

Estrogen receptor localization in the female genital tract.

The authors have used monoclonal estrophilin antibodies and the peroxidase antiperoxidase technique to characterize the distribution of estrogen receptor in the human vagina, uterus, and fallopian tubes. Exclusively nuclear localization of estrogen receptor was observed in epithelial cells, stromal cells of the lamina propria, and smooth muscle cells with both immunohistologic studies and immunoelectron-microscopy studies. The endometrium in various regions of the uterus (uterine isthmus, corpus, and fundus) was stained for estrogen receptor, with similar staining intensities in each of the respective cell types. There was no systematic regional variation in the staining intensity or distribution of cells with stained nuclei. The functionalis of the endometrium showed distinctive variation in the intensity of the staining for estrogen receptor with the menstrual cycle. The staining intensity of both endometrial epithelium and stroma was greatly reduced in the functionalis during the secretory phase. The vaginal epithelium did show some variation with the menstrual cycle, but it was much less than that observed in the functionalis of the endometrium. The basal cells of the vaginal stratified squamous epithelium, strongly stained during the proliferative phase, were less strongly stained during the secretory phase. No variation in the staining intensity for estrogen receptor was observed in different regions of the fallopian tube (isthmus, ampulla, and infundibulum), and the staining intensity varied only minimally with the menstrual cycle. The serosa of the female reproductive tract, connective tissues in the muscularis and in the vicinity of blood vessels, as well as neutrophils, eosinophils, mast cells, and lymphoid cells in the female genital tract were not stained for estrogen receptor.

Antibodies, Monoclonal↗
PubMed1986

Distribution of peroxidase and granulocytes in the human uterus.

A variety of uterine cell types demonstrate endogenous peroxidase activity. Ultracytochemical localization, biochemical assays, and uterine granulocyte counts were used to characterize peroxidase activity in various regions of the human uterus and cervix during the menstrual cycle and during the postmenopausal period. Previous studies of rat uteri, using electron microscopy and biochemical assays, have shown that endometrial peroxidase is induced by estrogenic stimulation (Anderson, De Sombre, and Kang, J Cell Biol 64:668, 1975; and Biol Reprod 16:409, 1977). Tissue samples from four regions of the human uterus and one sample from the endocervix were processed for ultrastructural cytochemistry, biochemical assay, and histology. Endogenous peroxidase activity was identified with electron microscopy in the endoplasmic reticulum of endometrial epithelial cells lining four regions of the uterine cavity; the isthmus, body (2), and fundus, of some proliferative phase (2 of 6), all secretory phase (4 of 4) and all postmenopausal (3 of 3) endometria. Peroxidase activity was not demonstrable in endocervical epithelial cells. Endogenous peroxidase activity was also identified in the cytoplasmic granules of uterine eosinophils and neutrophils and in the endoplasmic reticulum of mast cells. These uterine granule-containing cells, identified with special stains in the histologic sections, were quantitated. Approximately 80% of these "uterine granulocytes" from normal uteri without intrauterine devices were neutrophils. In women of reproductive age the uterine granulocytes, although present throughout the menstrual cycle, were most numerous in the endocervix and lower uterine segment. The highest biochemical assays of peroxidase activity were also obtained in the cervix and lower uterine segment. Uterine granulocyte counts varied directly with biochemical assays of peroxidase activity indicating that they were a major determinant of biochemical peroxidase activity. Endometrial epithelial peroxidase is anatomically and temporally well placed to function as an important adjunct in maintaining a mucosal barrier to microorganisms.

Adult↗
PubMed1985

Studies of the putative transforming protein of the type I human T-cell leukemia virus.

The putative transforming protein of the type I human T-cell leukemia virus (HTLV-1) is a 40-kilodalton protein encoded by the X region and is termed p40XI. On the basis of both subcellular fractionation techniques and immunocytochemical analysis, it is now shown that p40XI is a nuclear protein with a relatively short half-life (120 minutes). It is synthesized de novo in considerable quantities in a human T-cell line infected with and transformed by the virus in vitro, and it is not packaged in detectable amounts in the extracellular virus.

Antigens, Polyomavirus Transforming↗