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Biomedical subjects

P Monaghan

Publications and source records attributed to P Monaghan.

At least 91 records · Page 5Linked to original sources
PubMed1991

Positional cloning and characterization of a paired box- and homeobox-containing gene from the aniridia region.

Based on the map location of the aniridia (AN) locus in human chromosomal band 11p13, we have cloned a candidate AN cDNA (D11S812E) that is completely or partially deleted in two patients with AN. The less than 70 kb smallest region of overlap between the two deletions encompasses the 3' coding region of the cDNA. This cDNA, which spans over 50 kb of genomic DNA, detects a 2.7 kb message specifically within all tissues affected in AN. The predicted polypeptide product possesses a paired domain, a homeodomain, and a serine/threonine-rich carboxy-terminal domain, structural motifs characteristic of certain transcription factors. The concordance between expression and pathology, map location, structure, and predicted function argues that the cDNA corresponds to the AN gene.

Amino Acid Sequence↗
PubMed1991

Growth and development of the human infant breast.

Seventy-two samples of infant breasts, aged from newborn to 2 years, were collected at necropsy. Whole-mount preparations and histological sections were made. A system of classification was devised to study the extent of the structural development of the ductal system (morphological types I, II, and III) and the functional differentiation of the lining epithelium (functional stages I to V). There was no correlation between the age of the infant and the type of development of the ductal system. In contrast, the epithelial differentiation followed a chronological pattern, starting with secretory changes and apparently going through a period characterized by apocrine metaplasia before post-secretory involution. These epithelial changes were not associated with the morphological type of the ductal system. There were no distinguishing features between the breasts from the two sexes. Immunoperoxidase staining for actin and kappa-casein was carried out to study the myoepithelial cells and secretory cells, respectively. Myoepithelial cells were present at all stages and prominent staining for casein was observed up to 2 months of age. Embryonic-type adipose tissue was seen in 7 cases, in one of which it was associated closely with the developing ductal system. Extramedullary hematopoiesis was observed in the periductal connective tissue until 4 months of age. This paper describes the most extensive anatomical and histological study of the human infant breast to date and lays the foundation for a detailed study of the epithelial and stromal changes that take place during human breast development.

Actins↗
PubMed1991

Optimising technetium 99m diethylene triamine penta-acetate lung clearance in patients with the acquired immunodeficiency syndrome.

Pneumocystis carinii pneumonia (PCP) has become a major cause of morbidity and mortality due to infectious diseases, largely as a result of the acquired immunodeficiency syndrome (AIDS) epidemic. Since the mortality from recurrent infection is between 40% and 60%, early diagnosis and therapy are the keys to survival. The role of technetium 99m diethylene triamine penta-acetate (DTPA) aerosol pulmonary clearance was studied in 81 patients with AIDS. The mathematical technique of curve stripping was found to be the optimal method of analysis and to provide an overall sensitivity of 94% for the detection of interstitial pneumonitis. The procedure was superior to standard pathology parameters and radiography and more convenient than gallium 67 scintigraphy.

Acquired Immunodeficiency Syndrome↗
PubMed1991

Growth factor stimulation of proliferating cell nuclear antigen (PCNA) in human breast epithelium in organ culture.

Normal human breast explants were maintained in serum-free culture for 7 days in the presence of either insulin hydrocortisone and cholera toxin (I/H/CT), epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha). Explants were labelled with [3H] thymidine, fixed in methacarn and processed for autoradiography. Parallel sections were immunolabelled with anti-PCNA antibody and analysed with a CAS 200 image analyser. Thymidine labelling index (TLI) and PCNA expression produced similar results with both indices increased in response to I/H/CT, EGF and TGF-alpha. In sections double labelled for PCNA and autoradiography the majority of labelled cells were positive for both markers.

Autoantigens↗
PubMed1991

Characterization in vitro of luminal and myoepithelial cells isolated from the human mammary gland by cell sorting.

Luminal and myoepithelial cells have been separated from normal adult human breast epithelium using fluorescence activated cell sorting. Their isolation was based on the exclusive expression of two surface antigens, epithelial membrane antigen (EMA) and the common acute lymphoblastic leukaemia antigen (CALLA/CD10/neutral endopeptidase 24.11). Sorted luminal and myoepithelial cells displayed distinctively different morphologies when maintained in monolayer culture, differences which were enhanced by the addition of hydrocortisone, insulin and cholera toxin to the culture medium. The EMA-positive cells formed an attenuated monolayer with indistinct cell boundaries while CALLA-positive cells, by contrast, formed tightly packed arrays of refractile cells. The distribution of the cell type-specific markers cytokeratin 18 (luminal cells) and smooth muscle alpha-actin (myoepithelial cells) indicated that the sorted populations were approximately 98% pure. However, a significant minority (approximately 15%) of sorted luminal cells consistently expressed the basal-cell marker cytokeratin 14 in culture. A marked difference was noted in the proliferative behaviour of the two types of sorted cells, with myoepithelial cells dividing rapidly in response to the humoural additives, in contrast to the luminal cells which proliferated slowly. Both types of sorted cells could be cloned in the presence of feeder layers of mouse fibroblasts. Clones of luminal and myoepithelial cells were also distinctive; all "spread" luminal clones were similar in appearance to each other, although some cellular heterogeneity, including squamous metaplasia, was observed in "compact" myoepithelial clones. Both types were shown to have retained their original surface markers and to exhibit different cytoskeletal antigenic phenotypes when they were re-analysed after a 3-week growth period. Both spread and compact phenotypes were obtained when separately isolated ducts and alveoli were cloned. This detailed characterization of cells isolated from the human breast epithelium by flow cytometry provides the basis for further studies of luminalmyoepithelial interactions and growth responses of purified cell types in vitro.

Antigens, Differentiation↗
PubMed1991

Recent advances in electron immunocytochemistry.

Considerable progress has been made in the development of methods for the localisation of antigens at the ultrastructural level using both transmission and scanning electron microscopy. There is currently no technique which is appropriate for all antigens, although, almost without exception, colloidal gold-linked reagents provide the electron-dense marker. For post-embedding techniques, several approaches are available all utilising low temperatures during the preparative stages. Cryosectioning and embedding by the PLT method both require the antigen to withstand some degree of fixation. For fixation-sensitive antigens, freeze substitution in solvents lacking fixative agents is an approach offering considerable potential.

Frozen Sections↗
PubMed1991

An improved radionuclide technique for the detection of altered pulmonary permeability.

Technegas, an ultra-fine dry aerosol with prolonged retention in the lungs, can be modified by altering the atmosphere in which the carbon particles are generated. The modified Technegas has much faster clearance from the lung. The half-time pulmonary clearances with modified Technegas were compared to those obtained with conventional 99mTc DTPA aerosol in 50 patients. Interstitial lung disease was suspected in 12 while 38 were infected with the human immunodeficiency virus and suspected of having opportunistic lung infection. In 22 nonsmokers in whom no evidence of active pulmonary pathology was demonstrable, the mean half-time with DTPA was 52.5 min whereas the mean half-time with modified aerosol was 10.1 min. The mean half-time in 14 smokers in whom there was also no evidence of active pulmonary disease was 28.3 min with DTPA and 7.0 min with the modified method. In the 14 patients in whom altered pulmonary permeability was demonstrated by a short DTPA half-time (mean 4.8 min) there was also an accelerated half-time with modified Technegas (mean 2.5 min). It is concluded that the modified Technegas procedure offers a simple but accurate method of identifying individuals having opportunistic infection or other diffuse lung pathology.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Epidermal growth factor receptors and EGF-responsiveness of the human breast-carcinoma cell line PMC42.

PMC42 is an early-passage, well-differentiated, pleiomorphic human breast-cancer cell line. It expresses an average of 2.4 x 10(5) EGF receptor (EGFR) sites per cell when cultures are assayed using a radioreceptor assay. This is a relatively high figure for breast-carcinoma cell lines. Its responses to epidermal growth factor (EGF) were analyzed morphologically and by flow cytometry and immunocytochemistry. PMC42 responded in vitro to EGF with morphological changes, inhibition of "doming" and an increase in DNA synthesis in serum-containing medium in confluent or near-confluent cultures. When EGF receptor sites were localized immunocytochemically in PMC42, they were found to be variably expressed on the membrane of cells grown in the absence of EGF, with discrete areas of strong and weak cells of different morphological appearances; flow cytometry indicated a 10-fold range in EGFR levels between individual cells. Cultures maintained in the presence of EGF were less heterogeneous in their morphology and had a predominantly cytoplasmic, relatively uniform localization of EGFR with down-regulation of membrane EGFR levels. EGF-stimulated cultures were stained simultaneously for the presence of EGFR and the proliferation-related antigen Ki-67 and analyzed by both flow cytometry and immunocytochemistry. No correlation was observed between Ki-67 positivity and level of EGFR expression by individual cells. Electron microscopic localization of receptor sites demonstrated that the receptors were concentrated predominantly on the lateral and basal membranes. These results show that the PMC42 line reflects a number of the properties of heterogeneous breast tumours in situ. It may also continue to express growth-factor receptors in a manner that reflects their topographical distribution on normal luminal breast epithelial cells.

Antibodies, Monoclonal↗
PubMed1990

Expression of the CD34 gene in vascular endothelial cells.

All seven of a set of CD34 monoclonal antibodies that recognize epitopes on an approximately 110 Kd glycoprotein on human hemopoietic progenitor cells also bind to vascular endothelium. Capillaries of most tissues are CD34 positive, as are umbilical artery and, to a lesser extent, vein, but the endothelium of most large vessels and the endothelium of placental sinuses are not. Angioblastoma cells and parafollicular mesenchymal cells in fetal skin are also CD34 positive, as are some stromal elements. An approximately 110 Kd protein can be identified by Western blot analysis with CD34 antibodies in detergent extracts of freshly isolated umbilical vessel endothelial cells, and CD34 mRNA is present in cultured umbilical vein cells as well as other tissues rich in vascular endothelium (breast, placenta). These data indicate that the binding of CD34 antibodies to vascular endothelium is to the CD34 gene product, and not to crossreactive epitopes. Despite the presence of CD34 mRNA in cultured, proliferating endothelial cells, the latter do not bind CD34 antibodies. In addition, CD34 antigen cannot be upregulated by growth factors. We conclude that under these conditions, CD34 protein is downregulated or processed into another form that is unreactive with CD34 antibodies. Electron microscopy of umbilical artery, breast, and kidney capillary vessels reveals that in all three sites, CD34 molecules are concentrated on membrane processes, many of which interdigitate between adjacent endothelial cells. However, well-established endothelial cell contacts with tight junctions are CD34 negative. CD34 may function as an adhesion molecule on both endothelial cells and hematopoietic progenitors.

Antibodies, Monoclonal↗
PubMed1990

Peripubertal human breast development.

The microanatomical and histological appearance of the human breast has been studied during puberty. The macroscopic architecture of the mammary epithelial tree was identified and correlated with the histological appearance of material excised from defined regions of the breast preparations. Between ages 13 yrs and 15 yrs the human breast shows evidence of ductal elongation and branching, with lobules formed by lateral and dichotomous branching. The majority of ducts are lined by a two-layered epithelium consisting of recognisable myoepithelial and luminal cells. Less-well-defined multilayered regions were also observed in some areas, apparently at the site of lateral branching or early lobular development.

Adolescent↗
PubMed1990

Immunohistochemical distribution of c-erbB-2 in in situ breast carcinoma--a detailed morphological analysis.

An immunohistochemical study of c-erbB-2 expression was carried out on in situ (non-invasive) breast carcinoma, using antibody 21N, raised to the intracytoplasmic domain of the c-erbB-2 oncogene product. Strong membrane staining was observed in 44 out of 74 (59 per cent) cases of ductal carcinoma in situ (DCIS), but none of 48 lobular carcinoma in situ (LCIS) lesions. A detailed comparative morphological evaluation using several different parameters, including histological subtypes, was performed within the DCIS group. The results showed that there was a significant correlation between c-erbB-2 expression and the presence of large cell size, periductal lymphoid cell infiltration, marked nuclear pleomorphism, multinucleation, and a high mitotic rate. Of these, cell size appears to be the most important predictor of c-erbB-2 status, followed by the presence of periductal lymphoid cell infiltration. These results indicate, firstly, that LCIS and DCIS are biologically (as well as histologically) different and, secondly, that a subgroup of DCIS, which is associated with c-erbB-2 over-expression, exists and appears to have distinct histological features. The subgroup of DCIS cases which over-express c-erbB-2 may be a biologically definable category with prognostic importance. These results may therefore have relevance to breast screening programmes, but a larger study incorporating clinical data would be necessary to correlate these findings with clinical outcome.

Analysis of Variance↗
PubMed1990

Freeze-substitution without aldehyde or osmium fixatives: ultrastructure and implications for immunocytochemistry.

Cryo-fixation followed by freeze-substitution without aldehyde or osmium fixation has been investigated as a method for preparing biological specimens with a view to minimizing antigenic alteration. Samples of both solid tissues (mouse small intestine and human kidney) and a human tumour cell line grown in vitro were rapidly frozen by impact (slammed) onto a copper block cooled with liquid nitrogen. They were freeze-substituted at -80 degrees C in methanol, and embedded at low temperature in Lowicryl K4M or HM20. Resin blocks were polymerized by ultraviolet light. Well-preserved ultrastructure was observed in the outer 10-15 microns of all samples. Positive immunocytochemical localization of fixation-resistant and fixation-labile antigens was obtained on sections of human kidney and the human breast tumour cell line ZR-75-1 at both light and electron microscope levels.

Animals↗
PubMed1990

Role of bone scintigraphy in the early diagnosis of discitis.

Nine cases of recorded discitis are presented, in which scintigraphy played an important part in the diagnosis. The patients (five men (aged 21-75 years) and four women (aged 40-73 years)) had a history of back pain varying in duration from two days to three months. Final diagnosis was confirmed microbiologically (seven patients) or radiographically (two patients). Bone scintigraphy was a valuable diagnostic procedure for discitis with earlier detection than plain radiography in three patients and similar initial detection to that of third generation computed tomography. Single photon emission computed tomographic imaging increased diagnostic confidence by indicating the involvement of the adjacent vertebral bodies rather than of the pedicles or spinous processes.

Adult↗
PubMed1989

Expression of the common acute lymphoblastic leukaemia antigen (CALLA) in the human breast.

A biochemical and immunohistological study has been carried out to characterize the antigen in human breast reacting with antibodies to the common acute lymphoblastic leukaemia antigen (CALLA). Four different monoclonal antibodies to the CALLA antigen all stain the membrane of adult human myoepithelial cells. Surface labelling studies of freshly prepared human breast cells demonstrate that the anti-CALLA antibody, J5, immunoprecipitates a 100 kDa protein that co-electrophoreses with the CALLA antigen identified in the leukaemia cell line NALM-6. These results indicate that the CALLA antigen is expressed on myoepithelial cells and that the staining is not due to reactivity with a shared epitope on an unrelated molecule.

Antibodies, Monoclonal↗
PubMed1989

Isolation and properties of cell lines from the metastasising rat mammary tumour SMT-2A.

A new cell line Rat mammary (Rama) 900 was isolated from the ascitic version of the SMT-2A metastasising rat mammary tumour by stepwise adaptation of the tumour cells to tissue culture. The cells grew mainly as loosely-adherent aggregates, and were dependent during the first 18 passages in vitro on a feeder layer of mesothelial-like cells (Rama 950) obtained from the same tumour. Subcutaneous injection of Rama 900 cells in fat pads of syngeneic Wistar Furth rats yielded anaplastic primary tumours and extensive, gross metastases including those in lungs, lymph nodes, liver and bones, similar to the parental transplantable tumour. The extent of metastatic spread from subcutaneous fat pads was increased by passage 17 in vitro for the Rama 900 cells. A similar extent of metastatic spread was achieved at earlier times by injecting the original cells with the non-tumorigenic Rama 950 cells in vivo. Subcutaneous injection of Rama 900 into thymectomised rats or MF1 nu/nu mice yielded fewer tumours, most of which regressed. No metastases occurred in the thymectomised rats and fewer metastases, mainly in lungs but not in lymph nodes, livers or bones, were seen in the nude mice. The ascitic tumours formed by intraperitoneal injection of nude mice contained both anaplastic rat cells similar to Rama 900 and mouse mesothelial-like cells similar to Rama 950. Although these anaplastic ascites cells failed to yield any tumours in syngeneic or thymectomised rats, they still produced tumours and metastases, including those in lymph nodes, in nude mice.

Animals↗