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Biomedical subjects

A Lewis

Publications and source records attributed to A Lewis.

At least 181 records · Page 10Linked to original sources

Role of surgery in the treatment of pseudomyxoma peritonei.

Laparotomy was undertaken in five patients with pseudomyxoma peritonei presenting over a 9-year period. Three patients remain alive and have not required further surgery. A fourth patient has died of an unrelated condition, having been asymptomatic for 8 years, and the fifth has succumbed to the disease 20 years after presentation. It is suggested that an aggressive approach to initial surgery, and in particular thorough peritoneal toilet, carries the best prospects of long-term survival.

Adult↗

Membrane receptors, involved in up-regulation of calcium channels in bovine adrenal chromaffin cells, chronically exposed to ethanol.

Release of catecholamines, induced by carbachol, from bovine adrenal chromaffin cells, maintained in culture, is potently inhibited by the presence of ethanol (50 mM). This inhibition is prevented by the concomitant presence of bicuculline (1 microM) or by the inverse agonist at the benzodiazepine receptor, Ro 15 4513 (50 nM), arguing that the effect of ethanol is, at least, partly due to potentiation of endogenous GABA at the GABAA receptor sites on these cells. Exposing cells to medium containing ethanol (200 mM) for 4 days results in an approximately 100% increase in binding sites for [3H]dihydropyridine on membranes of adrenal chromaffin cells. This increase in binding sites was largely prevented by the presence in the culture medium of Ro 15 4513 (50 nM), the dihydropyridine calcium channel agonist BAY K 8644 (50 nM) or the calcium channel antagonist, nitrendipine (50 nM). The results strongly suggest that the increase in binding sites for [3H]dihydropyridine represents an adaptation of cells to overcome the inhibitory effect of ethanol on the excitability of cells (which is, at least partly, due to some action at the GABAA receptor). The protein containing the receptor site for dihydropyridine drugs is clearly also involved in controlling its up-regulation by ethanol, but this is probably not directly related to its channel function, since both activators and inhibitors of the dihydropyridine-sensitive channel prevented ethanol-induced up-regulation of binding sites for [3H]dihydropyridine.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

New versatile restraining device for use with rats in pharmacokinetic or pharmacological studies involving chronically implanted cannulae or electrodes.

A new design for a rat restraining device is described. Features of the device include facilities to allow the accommodation of rats of a very wide weight range and to maximize its convenience in use in a variety of procedures including the collection of blood, feces, and urine. The restrainer is designed to minimize stress in the rat in the course of pharmacokinetic or pharmacological studies. The device is particularly useful for studies involving chronically cannulated animals.

Animals↗

Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: relation to in vivo phenotypes and influence of medium.

The luminal and basal epithelial cells in the human mammary gland can be distinguished in tissue sections on the basis of the pattern of keratins they express. Moreover, the invasive cells in primary carcinomas show a keratin profile that corresponds to that of the dominant luminal cell (7, 8, 18, 19). When homogeneous populations of luminal epithelial cells from milk or from breast cancer metastases are cultured the profile of keratin expression seen in vivo is maintained. We have therefore used monospecific antibodies reactive with individual keratins to examine the phenotype of cells cultured in three different media from reduction mammoplasty tissue that contains both luminal and basal cells. The phenotype of cells cultured from primary breast cancers in one of these media (MCDB170) has also been examined. In characterizing cell phenotypes, antibodies to a polymorphic epithelial mucin (PEM) expressed in vivo by luminal cells, and to smooth muscle (a) actin, expressed in vivo by basal cells, have also been used. Our results show that proliferation of different cell phenotypes is selected for in different media. In milk mix (MX) developed for growth of luminal cells from milk, only the luminal cell phenotype proliferates (for only 1 or 2 passages). In medium MCDB 170, which was developed for long-term growth of human mammary epithelial cells from reduction mammoplasty organoids, cells from the basal layer proliferate, while in MM medium the basal phenotype dominates, but a few cells with the luminal phenotype are found. Around passage 3, in medium MCDB 170, most cells senesce and a subpopulation of cells proliferates on further passage. These cells retain expression of the basal epithelial keratins but also express some features characteristic of luminal epithelial cells, suggesting that the basal layer may contain a stem cell that can develop along the luminal lineage. In culture, however, they do not express keratin 19, which in vivo is a feature of the fully differentiated luminal cell. The cells cultured from primary breast cancer in medium MCDB 170 have a similar keratin profile to that of the normal cells cultured in this medium. They do not express keratin 19, even though the invasive cells in primary cancers homogeneously express this keratin in vivo. The invasive phenotype, which in its keratin profile corresponds to the differentiated luminal cell and that of the metastatic cancer lines, cannot be cultured from primary breast cancers using MX, which supports proliferation of the corresponding normal cell.

Breast↗

Palisaded encapsulated neuromas. A clinicopathologic study.

Reed et al described the clinical and light-microscopic findings of palisaded encapsulated neuromas in 1972, but few cases have been reported since. We have studied 81 consecutive tumors. Clinically, these were solitary, asymptomatic, 2- to 6-mm, flesh-colored papules, usually located on the face of middle-aged patients. The correct diagnosis was rarely made; the lesion was most often mistaken for a basal cell epithelioma, melanocytic nevus, or other benign tumor. Light microscopy revealed single or multiple encapsulated dermal lobules composed of interlacing Schwann cells. Variable numbers of fine axons and myelin sheath remnants were present. Palisading of nuclei was not a prominent feature. Electron microscopy demonstrated substantial numbers of class C fibers (mostly nonmyelinated) only partially enveloped by Schwann cell cytoplasm. Pathologically, palisaded encapsulated neuromas are distinctive true neuromas resembling those seen in the multiple mucosal neuroma syndrome. Electron-microscopic findings are similar to those seen in peripheral nerve regeneration, suggesting that palisaded encapsulated neuromas may be traumatic in origin, and could represent regeneration following local minor injury to the skin.

Adult↗

Evidence for light-induced lysine conformational changes during the primary event of the bacteriorhodopsin photocycle.

Fourier transform infrared difference spectroscopy is used to examine the role of lysine in the primary event of the bacteriorhodopsin photocycle. Isotopically labeled lysine is used to tentatively assign the lysine modes in the BR and K species. The results suggest that the lysine side-chain undergoes conformational changes in concert with the known light-induced chromophore structural alterations.

Bacteriorhodopsins↗

Nonlinear optical properties of potential sensitive styryl dyes.

The nonlinear optical properties of dyes that alter their optical characteristics rapidly with membrane potential are described. The second harmonic signals from these dyes characterized in this paper are among the largest that have been detected to date. Structural conclusions are drawn from the second harmonic signals generated by the Langmuir Blodgett monolayers used in these measurements. Our results indicate that with appropriate instrumentation second harmonic signals could readily be detected from living cells stained with these dyes.

Cell Membrane↗

Development and characterization of breast cancer reactive monoclonal antibodies directed to the core protein of the human milk mucin.

A mucin molecule, which has a molecular weight of greater than 400,000 and which carries tumor associated epitopes recognized by monoclonal antibodies HMFG-1 and HMFG-2, has been purified from human skimmed milk by affinity chromatography followed by passage through a size exclusion column. While treatment of the mucin with hydrogen fluoride for 1 h at 4 degrees C removed the peripheral oligosaccharides, treatment with HF for 3 h at room temperature removed all of its lectin binding ability and revealed a dominant polypeptide of about 68,000. This appears to be the size of the mucin core protein. Monoclonal antibodies have been developed that react with the stripped and partially stripped molecule but not with the intact mucin. From the initial screening on histological sections one of these antibodies, SM-3, reacts with 91% of breast carcinomas but shows little or no reactivity on benign mammary tumors, normal resting, pregnant, or lactating breast. It appears that this monoclonal antibody is reacting with an epitope that is usually masked by oligosaccharide moieties in normal cells but which is exposed, perhaps due to aberrant glycosylation, in malignant cells.

Amino Acids↗

Immunological analysis of mucin molecules expressed by normal and malignant mammary epithelial cells.

Many existing MAbs raised against the human milk fat globule or against carcinoma cells recognise epitopes on high-molecular-weight glycoproteins. In a comparative ELISA assay a number of these antibodies have been shown to react with an extract of the human milk fat globule. In comparative immunoblots of cultured normal milk cells and breast cell lines, all were found to bind to large molecules which show some variation in molecular weight depending on the cell source. The HMFG-2 antibody, which is widely used in cancer diagnosis, also recognises epitopes on lower-molecular-weight components. In T47D cells these may be as small as 80,000 Mr, and with electron microscopy this cell line can be shown to accumulate HMFG-2-reactive components in the Golgi apparatus. Using an HMFG-2 affinity column we have immunopurified HMFG-2-reactive material from the 2 breast cancer cell lines MCF-7 and T47D and shown that all of the above antibodies react in a solid-phase ELISA with the purified material. In addition to high-molecular-weight components, the immunopurified material was found to contain lower-molecular-weight components including a glycoprotein of 68,000 Mr that was not, however, recognised by the HMFG-2 antibody on a Western blot. We have used this immunopurified material to generate new MAbs. All of these recognise the high-molecular-weight bands seen with the other antibodies, but 2 of them also recognise a band at 68,000 Mr in blots of MCF-7 and T47D. The second-generation antibodies show a spectrum of reactivity on tissues similar to HMFG-2 and one reacts at least as strongly as HMFG-2 with methanol-acetone-fixed sections of breast cancers.

Antibodies, Monoclonal↗

Levels of bovine papillomavirus RNA and protein expression correlate with variations in the tumorigenic phenotype of hamster cells.

Three independent cell lines were established from primary cultures of LSH hamster embryo cells infected with bovine papillomavirus type 1 (BPV-1). Although these cell lines differed in their in vitro saturation densities, none was capable of colony formation in soft agar. Interestingly, two cell lines (BPV-HE1 and BPV-HE3) were tumorigenic in nude mice, syngeneic hamsters, and allogeneic hamsters, whereas BPV-HE2 was not. All three cell lines contained similar numbers of the BPV-1 genome (approximately 50 to 200 copies per cell). However, the nontumorigenic BPV-HE2 cell line contained very low levels of BPV-specific RNA and only small amounts of the BPV-1 E5 transforming protein. The efficiency and rate of tumor formation by BPV-HE1 and BPV-HE3 correlated directly with the apparent amount of viral E5 protein. This analysis suggests that there is a threshold level of BPV protein synthesis required for tumorigenicity, there is a continuum of tumorigenic phenotypes which may depend upon the level of BPV protein expression, and BPV-transformed hamster cells can withstand allogeneic transplantation.

Animals↗

Fluorescence light microscopy of F-actin in retinal rods and glial cells.

The actin cytoskeleton of rod photoreceptors and glial cells in toad retina has been directly viewed using fluorescence microscopy of cells labeled with a potent phallotoxin that specifically binds to F-actin. The three-dimensional organization of this cytoskeletal protein consists of actin filaments, which course through the inner segment and end at the tips of the calycal processes surrounding the base of the outer segment. A transverse layer of actin staining is also observed at the base of rod outer segments in the region where new discs are formed. At the level of the external limiting membrane, evidence has been found for rings of actin within the glial cells that surround the photoreceptors. These actin rings form a structural meshwork in which photoreceptor cells are embedded.

Actins↗

Extended X-ray absorption fine structure of Mn2+ and Mn2+ X ATP complex bound to coupling factor 1 of the H+-ATPase from chloroplasts.

The spinach chloroplast ATPase, coupling factor 1, contains three tight Mn2+-binding sites which interact cooperatively. The bound manganese coordinations were studied by x-ray absorption fine structure analysis. Mn2+ was found to be bound to the enzyme with an average Mn-O bond length of 2.15 +/- 0.15 A, significantly shorter than the 2.15 +/- 0.15 A of the Mn-O bond of the average first hydration shell for Mn2+ in aqueous solution. On adding ATP to the manganese-enzyme mixture, a tertiary complex of Mn2+ X ATP X enzyme was formed as indicated by the appearance of a second shell. Mn-P bond distances were estimated at 4.95 +/- 0.15 A in the tertiary Mn2+ X ATP X enzyme complex, which was considerably longer than the Mn-P bond distance of 3.36 +/- 0.15 A for the Mn2+ X ATP complex in aqueous solution. The Mn-P bond distance in the tertiary Mn2+ X ATP X enzyme complex decreased to 4.32 +/- 0.15 A when selenite, a potent effector of ATPase activity, was added. Based on these results, it is suggested that the tertiary complex is required for catalysis. The stimulation of ATP hydrolysis by anions such as selenite may be the result of shortening the distance between Mn2+ and the ATP phosphates in the enzyme active site.

Adenosine Triphosphate↗