Malignant MCF10CA1 cell lines derived from premalignant human breast epithelial MCF10AT cells.

The MCF10 series of cell lines was derived from benign breast tissue from a woman with fibrocystic disease. The MCF10 human breast epithelial model system consists of mortal MCF10M and MCF10MS (mortal cells grown in serum-free and serum-containing media, respectively), immortalized but otherwise normal MCF10F and MCF10A lines (free-floating versus growth as attached cells), transformed MCF10AneoT cells transfected with T24 Ha-ras, and premalignant MCF10AT cells with potential for neoplastic progression. The MCF10AT, derived from xenograft-passaged MCF10-AneoT cells, generates carcinomas in approximately 25% of xenografts. We now report the derivation of fully malignant MCF10CA1 lines that complete the spectrum of progression from relatively normal breast epithelial cells to breast cancer cells capable of metastasis. MCF10CA1 lines display histologic variations ranging from undifferentiated carcinomas, sometimes with focal squamous differentiation, to well-differentiated adenocarcinomas. At least two metastasize to the lung following injection of cells into the tail vein; one line grows very rapidly in the lung, with animals moribund within 4 weeks, whereas the other requires 15 weeks to reach the same endpoint. In addition to variations in efficiency of tumor production, the MCF10CA1 lines show differences in morphology in culture, anchorage-independent growth, karyotype, and immunocytochemistry profiles. The MCF10 model provides a unique tool for the investigation of molecular changes during progression of human breast neoplasia and the generation of tumor heterogeneity on a common genetic background.

The hyperplasia-to-carcinoma sequence in the breast. Immunohistochemical-histologic correlations.

Breast cancer is probably the result of a series of genetic events, each with its own histopathologic correlate in the hyperplasia to carcinoma sequence. The expression of breast cancer markers in hyperplasia and tumors are well known, but few studies have investigated their sequential expression among hyperplastic and cancerous lesions within the same breast. Using breast tissue obtained from a single procedure, we correlated the immunohistochemical expression of several breast cancer markers with the histopathologic stage of proliferative breast disease. We selected 14 cases in which various degrees of hyperplasia coexisted with carcinoma. Serial sections were reacted with antibodies to DF3, c-erbB-2, p53 (DO7 and CM1), B72.3, and cyclin D1. We found that within an individual breast, the number of breast cancer markers expressed increased with progression from hyperplasia to atypical hyperplasia to carcinoma. Cytoplasmic DF3 was first expressed at the level of simple hyperplasia, followed by c-erbB-2 in atypical hyperplasia. Overexpression of p53 was confined to carcinomas, and thus appeared to be a late event. B72.3 was expressed in three carcinomas and in one atypical hyperplasia, although the associated carcinoma was negative. Carcinomas that expressed cytoplasmic DF3 and c-erbB-2 were associated with atypical hyperplasias that also expressed cytoplasmic DF3 and c-erbB-2, with one and two exceptions, respectively. No specific cyclin D1 staining pattern was observed.

Outpatient frozen sections by telepathology in a Veterans Administration medical center.

A relatively simple telepathology system is described for evaluating the margins of excision of cutaneous basal and squamous carcinomas. The system uses a microscope with a built-in television camera, but no eyepieces. The image is projected onto an adjacent monitor and transmitted by T1 line at 768 Kbs to a remote, large screen monitor. The microscope is operated by the surgeon under the telephone direction of the pathologist at the remote site. In a series of 66 cases involving more than 400 individual tissue blocks, we have had only 2 cases with false-negative interpretations and 2 in which the block was not fully displayed on the frozen section. In 15 cases, 1 or more surgical margins were positive, and the surgeon proceeded to excise additional tissue. Our success is attributed to dedicated involvement by the surgeon, very high-quality frozen sections, and the experience of the pathologist.

Contractile and arrhythmic effects of endothelin receptor agonists in human heart in vitro: blockade with SB 209670.

It is known that binding sites with endothelin(A) (ET)(A) and ET(B) receptor characteristics coexist in human heart but little is known about the receptors that mediate cardiostimulant effects of ET receptor agonists or their consequences. Functional studies were performed on isolated human cardiac tissues. The maximal positive inotropic effects of ET-1 were right atrium > left atrium = right ventricle. The rank order of potencies of agonists in right atrium was sarafotoxin S6c > ET-1 = ET-2 > or = ET-3. The ET(A) receptor-selective compounds BQ123 (10 microM) and A-127722 (1 microM) only slightly blocked (<0.5 log-unit shift) the effects of lower concentrations of ET-1, and the ET(B) receptor antagonist Ro46-8443 (10 microM) did not cause blockade. SB 209670 caused concentration-dependent rightward shifts of ET-1 and sarafotoxin S6c concentration-effect curves with Schild slopes not different from one and affinities (-logM K(B)) of 7.0 and 7.9, respectively. ET-1 caused arrhythmic contractions in right atrial trabeculae that were prevented by 10 microM SB 209670 but not 10 microM BQ123 or 1 microM A-127722, precluding ET(A) receptors. ET-1 caused a higher incidence of arrhythmic contractions in tissues taken from patients treated with beta-blockers before surgery than in tissues from non-beta blocker-treated patients. Sarafotoxin S6c produced arrhythmias that were prevented by SB 209670. The positive inotropic effects of ET-1 in human right atrial myocardium are mediated mostly by a non-ET(A), non-ET(B) receptor. Ventricular inotropic ET receptors differ from atrial inotropic ET receptors. ET-1 induced arrhythmic contractions in human atria do not appear to be mediated by an ET(A) receptor.

Progression of premalignant MCF10AT generates heterogeneous malignant variants with characteristic histologic types and immunohistochemical markers.

The MCF10AT premalignant human breast epithelial cells form benign ductal structures in immunodeficient mice which sporadically progress to carcinoma in situ and invasive cancers of different histologic types. MCF10CA1 cell lines are malignant variants derived by serially passing small pieces of tumors in athymic mice before establishing cells in culture. As these MCF10CA1 variants gave rise to heterogeneous tumors, some cell lines were cloned. Inoculated into immunodeficient mice, these variants produce squamous carcinomas with an undifferentiated component or adenocarcinomas also with an undifferentiated component. Immunohistochemistry utilized antibodies against DF3, c-erbB-2, cyclin Dl, m keratin, p keratin, p53, B72.3 and estrogen receptor. We detected characteristic patterns for squamous carcinomas, for adenocarcinomas, and for each undifferentiated component, that is the undifferentiated components of the squamous and glandular carcinomas were distinct. Only adenocarcinomas were focally ER positive. One uncloned variant that produced cancers with a glandular component, MCF10CA1h, was cloned and cells were injected into mice. This clone produced only undifferentiated carcinomas that, compared to tumors formed by the parental uncloned variant, had lost ER, DF3 and c-erbB-2 expression, but more strongly expressed p53. Our data demonstrate the potential of the premalignant MCF10AT model to generate heterogeneity, including both estrogen receptor-positive as well as estrogen receptor-negative tumors, during progression.

The original illustrations of Hodgkin's disease.

In this report, the illustrations from the original papers on Hodgkin's disease are used to trace its early history. Thomas Hodgkin's report included six cases of his own and a seventh given to him by Robert Carswell, whose beautiful colored pictures of the latter case accompanied Hodgkin's presentation. Early clinical pictures are also presented. The histologic definition of the disease, with its characteristic cell, is traced with drawings from the reports of Greenfield, Sternberg, Reed, and Andrews. Modern histologic and immunocytochemical confirmation (Leu-M1 reactivity) of some of Hodgkin's original cases, preserved at Guy's Hospital, London, UK, are also illustrated. It is concluded that not only did Hodgkin describe cases of Hodgkin's disease that meet present-day criteria, he also included at least one case of non-Hodgkin's lymphoma, a term that now might be considered a misnomer.

Efficiency measurement of health care: a review of non-parametric methods and applications.

There has been increasing interest in measuring the productive performance of health care services, since the mid-1980s. This paper reviews this literature and, in particular, the concept and measurement of efficiency and productivity. Concerning measurement, we focus on the use of Data Envelopment Analysis (DEA), a technique particularly appropriate when multiple outputs are produced from multiple inputs. Applications to hospitals and to the wider context of general health care are reviewed and the empirical evidence from both the USA and Europe (EU) is that public rather than private provision is more efficient.

Schizophrenia and genetics: a review and critique for the psychiatric nurse.

Nursing authors such as McCrone and Gournay persist in the orthodox view that the genetic heritability of schizophrenia is an established scientific fact. In this article I take issue with the view they propound by way of an examination of the literature pertaining to schizophrenia and genetics. My reading of the literature suggests that the certainty they seek is not to be found in any of the scientific evidence so far put forward in support of the genetic hypothesis. Much of the early work in this field is fatally flawed and can no longer be used to support the genetic argument. More recent work tends to give increasing support to a greater role for environmental factors than is compatible with a hereditarian argument. Fifty years of research in this field has consistently failed to provide reliable scientific evidence in support of any of the genetic hypotheses relating to schizophrenia. Nurses, it is argued, should not be dispensing advice to patients on the basis of a genetic model that is at best mere hypothesis.

Altered expression of c-erbB-2, DF3, b72.3, p53 and Ki-67 with progression and differentiation to two distinct histologic types of invasive carcinoma in the MCF10AT human xenograft model of proliferative breast disease.

Human breast epithelial MCF10AT cells form simple ducts in nude/beige mice which eventually become hyperplastic and sporadically progress to carcinomas. Altered immunohistochemical detection of c-erbB-2, DF3, B72.3, p53 and Ki-67 was observed with progression and differentiation to two distinct histologic types of invasive carcinoma. c-erbB-2 and DF3 were detected in 50% and 18% of lesions at the stage of atypical hyperplasia and expression increased to 78% and 54% in invasive adenocarcinomas. In contrast, a group of six unusual undifferentiated tumors with squamoid features did not express c-erbB-2 or DF3, but both B72.3 (4/6) and p53 (6/6) were detected.

Patient-controlled epidural versus intravenous pethidine to supplement epidural bupivacaine after abdominal aortic surgery.

In a double-blind, randomized, crossover study of 25 patients after abdominal aortic surgery, we compared patient-controlled analgesia (PCA) with epidural versus intravenous pethidine. All patients received continuous epidural infusions of 0.125% bupivacaine adjusted to maintain appropriate sensory levels. The 48 hour study period commenced 36 to 48 hours after surgery and covered postoperative days 2 and 3. There was a crossover in PCA mode (epidural or intravenous) after 24 hours. Plasma pethidine concentration at the end of each 24 hour period and the total 24 hour pethidine dose did not change significantly between postoperative days 2 and 3. Pethidine plasma concentration was lower after 24 hours epidural than after intravenous PCA [125 (SD 108) ng/ml versus 171 (SD 107) ng/ml, P = 0.03], although pethidine dose did not differ significantly [mean 147 (SD 124) mg/24 h]. Visual analog pain scores (VAS) did not differ significantly between postoperative days 2 and 3, or at rest between epidural and i.v. groups. However, VAS with coughing and with abdominal palpation were lower in the epidural PCA group (P = 0.05, 0.008). With a background epidural infusion of 0.125% bupivacaine, PCA with epidural pethidine provided better pain control than PCA intravenous pethidine and this was achieved at lower plasma pethidine concentrations.

A personal reflection on the nature of somatic treatments and the implications for mental health nursing.

Somatic treatments in psychiatry, ECT (electro-convulsive therapy) and pharmacotherapy, are based on a particular conception of our nature as human beings, and on a particular conception of clinically constructed reality. Acceptance of the basis for such treatments constitutes a tacit admission of our nature as being essentially materialist and deterministic biological machines, and undermines the connectedness which ensures that we each acknowledge the humanity and humanness of the other. By objectifying the other as different (by reason of disordered biology), these connections are severed and allow the imposition of physical forms of treatment in the name of cure. In this personal reflection, I draw upon my own experience of ECT from a nursing perspective, and attempt to explain the basis of my antipathy for this treatment. In the course of this exploration I also raise some of the issues which I feel nursing must deal with if it is to continue uncritically to adopt the biological paradigm as the basis of its praxis.

Is there anyone in there? Psychiatric nursing meets biological psychiatry.

Mental health nursing operates without a clear idea of the nature of both mind and mental. Increasingly, this lack of a defensible theoretical position has led to an increasing dependence on the concept of mind now current in biological psychiatry. But the materialist monism of biological psychiatry is itself open to doubt, particularly concerning its dependence on a priori assumptions about the nature of the relationship between mind and brain. The reductionism and objectivism inherent in this approach necessarily ignores that aspect of mind most germane to nursing, the first-person nature of the mental. An alternative is briefly sketched which stresses the mediation of behaviour by the brain, rather than viewing behaviour as causally related to brain processes. Adoption of this approach conserves nursing's focus on the subjective experience as being paramount. Nursing education should, therefore, be wary of incorporating the biological approach without a critical analysis of its suppositions and of the conception of human nature which it supports.

The effects of cardiopulmonary bypass on total and unbound plasma concentrations of propofol and midazolam.

OBJECTIVE: To examine the effects of cardiopulmonary bypass (CPB) on total and unbound plasma concentrations of propofol and midazolam when administered by continuous infusion during cardiac surgery. DESIGN: Prospective clinical study. SETTING: University hospital. PARTICIPANTS: Twenty-four adult patients undergoing cardiac surgery. INTERVENTIONS: Patients received either propofol or midazolam to supplement fentanyl anesthesia. Twelve patients received a propofol bolus (1 mg/kg) followed by an infusion of 3 mg/kg/hr. A second group received midazolam, 0.2 mg/kg bolus, followed by an infusion of 0.07 mg/kg/hr. MEASUREMENTS AND MAIN RESULTS: Blood sample were collected from the radial artery cannula at 0, 2, 4, 8, 8, 10, 15, 20 minutes and then every 10 minutes before CPB, at 1, 2, 3, 4, 6, 10, 15, 20 minutes and then each 10 minutes during CPB. On weaning from CPB samples were collected at 0, 5, 10 and 20 minutes. Plasma binding, total and unbound propofol and midazolam concentrations were determined by ultrafiltration and high-pressure liquid chromatography (HPLC). CPB resulted in a fall in total propofol and midazolam plasma concentrations, but the unbound concentrations remained stable. The propofol unbound fraction increased from 0.22 +/- 0.06% to 0.41 +/- 0.17%. The midazolam unbound fraction increased from 5.6 +/- 1.0% to 11.2 +/- 2.1%. CONCLUSIONS: Unbound concentrations of propofol and midazolam are not affected by cardiopulmonary bypass. Total intravenous anesthesia algorithms do not need to be changed to achieve stable unbound plasma concentrations when initiating CPB.

Ozone does not react with human erythrocyte membrane lipids.

Ozone was applied to sealed red cell ghost membranes at the rate of 95 nmol/min for periods up to 20 min. Acetylcholine esterase, on the outer face of the membrane, was inhibited up to 20%. Glyceraldehyde-3-phosphate dehydrogenase, on the inner surface of the membrane, was inhibited up to 87%. These differences reflected the inherent susceptibilities of the two enzymes and the presence or absence of the membrane barrier. Analysis of the total lipids of the ozone-treated ghosts showed no significant change in the distribution of lipid classes and no significant change in the fatty acid composition. There was no significant change in the fatty acid composition of the phosphatidylcholine fraction. There was a slight increase in 18:0 and 20:2 + 20:3 in the phosphatidylethanolamine fraction. There was no change in the molecular species distribution of the phosphatidylcholine or the phosphatidylethanolamine fraction. There was no evidence for the formation of the phospholipid ozonolysis product, 1-acyl-2-(9-oxo-nonanyl) derivatives of glyceryl-phosphoryl choline. There was no decline in the amount of cholesterol in the lipids derived from ozone-treated red cell membranes. Treatment of red cell ghost membranes and, by implication, the plasma membrane of cells by ozone therefore oxidizes peripheral proteins before it oxidizes lipids.

Reaction of ozone with protein tryptophans: band III, serum albumin, and cytochrome C.

Treatment of red cell ghosts with ozone inhibited both AChE (marking the outside of the membrane) and G3PDH (marking the inside of the membrane). There was no change in tryptophan fluorescence of the ghosts after the ozone treatment. Band 3 protein was isolated from the ozone-treated ghosts. The protein was digested with trypsin to obtain water soluble peptides from the cytoplasmic N-terminal tail and the interhelical loops. Fluorescent peptides included GWVIHPLGLR from the outer loop between helices 7 and 8, and peptide WMEAAR from the N-terminal cytoplasmic tail. Neither one of these peptides was oxidized by ozone. This was true whether or not the ghosts were sealed. We conclude that the position of these tryptophans either in the membrane structure, or because of binding to other proteins in the cytoplasmic tail, protects them from oxidation by ozone. Treatment of horse heart cytochrome c with ozone did not change the absorbance spectrum in the heme region or the tryptophan absorbing region. HPLC of the ozone-treated cytochrome c showed that cytochrome c was being modified, indicated by a change in the elution time. Treatment of cytochrome c with ozone did not change the activity in the NADH-cytochrome c reductase assay. Digestion of the ozone-treated cytochrome c with trypsin gave peptides which demonstrated normal fluorescence. (Cytochrome c has abnormally low fluorescence, which is not changed by ozone exposure.) The peptides were separated by HPLC. The fluorescence of the tryptophan-containing peptide (GITWK) was not decreased by treatment of the cytochrome c by ozone. Amino acid analysis of the ozone-treated cytochrome c indicated that methionine was oxidized. We conclude that tryptophan in cytochrome c is protected from oxidation by ozone because of the interaction with the porphyrin ring. Bovine serum albumin and human serum albumin were treated with ozone. There was a monotonic decrease in tryptophan fluorescence in both cases. Digestion of BSA with trypsin produced two fluorescent peptides. The peptide FWGK was identified by coelution with the authentic peptide. The putative peptide AWSVAR was not the same as the chemically synthesized peptide. The peptide sequences FWGK and "AWSVAR" were both oxidized in ozone-treated bovine serum albumin, with no detectable discrimination. Tryptic digestion of the ozone-treated human serum albumin produced a single fluorescent peptide, which was oxidized by ozone. The putative peptide AWAVAR in the tryptic digest of HSA was distinct from chemically synthesized peptide. The oxidation of tryptophans in proteins by ozone is markedly influenced by position in tertiary structure, position in membrane structure, and by chemical interactions within the protein.

A reply to Goddard's 'spirituality as integrative energy'.

Nancy Goddard's 'spirituality as integrative energy' serves to highlight the importance of spirituality, while at the same time seeking to transform its nature. In this reply I take issue with the concept of spirituality as integrative energy, arguing that such a reconceptualization effectively strips spirituality of its meaning in the name of a spurious scientism. I argue that science is not the only valid mode of knowledge, and that, indeed, the scientific way of knowing may be particularly inappropriate in the context of the spiritual sphere I briefly consider the gradual separation of spirit from body in the development of western medical science, and point out the commonalities between the scientific and spiritual modes of apprehending the world. While it is important that nurses be aware of the spiritual dimension, I argue that this awareness should not be purchased by reducing spirituality to more 'acceptable' 'scientific' terms, but rather that nurses should be afforded the opportunity to study the concept in its historical, literary and philosophical context.

A reply to Kevin Gournay's 'Schizophrenia: a review of the contemporary literature and implications for mental health nursing theory, practice and education'.

Mental health nurses are being increasingly encouraged to move to a more biologically oriented approach. Authors such as Professor Gournay promulgate the myth that biological psychiatry will soon be able to provide an unambiguous model of the nature of mental disorder. A closer examination of some of the biological research identified by Gournay as moving us closer to this ideal reveals, however, the confused and unconvincing nature of the results so far achieved. In the light of this it seems premature to be advocating wholesale acceptance of the biological model by nursing. Nurses should be made aware of the investigations of the biologists, but, unless they are equipped with the necessary intellectual tools to place such investigations in context, they will continue to be susceptible to the rhetoric of proselytisers such as Gournay.