PubMed HealthSearch

Biomedical subjects

A L Latner

Publications and source records attributed to A L Latner.

At least 19 recordsLinked to original sources

Analyses of protein extracts of human breast cancers: changes in glycoprotein content linked to the malignant phenotype.

The protein and glycoprotein composition of Triton X-100 extracts of breast biopsies from 17 women with benign breast disease and from 11 women with invasive breast carcinoma were investigated using electrophoresis in SDS-containing gradient polyacrylamide gels, followed by Coomassie Blue (CB) staining and the binding of radio-iodinated wheat germ agglutinin (WGA). Patterns were analysed after the CB-step for differences in protein composition, and after the WGA-step for differences in glycoprotein composition. Tissue extracts from patients with benign breast disease have less CB stained bands than similar extracts from the cancer patients. A particular consistent change was the appearance of an extra band at 58 Kdaltons in the cancer extracts. In contrast to the CB results, WGA detected less major bands, in the 40-60 Kd region, in the cancer extracts than at similar locations in benign extracts. Analysis of blood sera using the above techniques suggested that certain serum proteins could account for some of the WGA changes, but not the changes after CB staining. However, residual contamination of the specimens by blood proteins seemed unlikely because of the washing procedure used, unless these components were very strongly associated with the tissue. Differential synthesis of serum proteins by benign and malignant breast tissue may also explain some of our findings. Examination of the histopathology adjacent to the extracted tissue suggested that the degree of reduction in WGA-binding may be related to the extent of local invasiveness. Other animal and human studies suggest that reduced glycosylation of tumour-associated proteins may be linked to increased malignancy. The current findings may reflect a general pattern of change in tumour glycoprotein composition linked to malignant expression.

Adult

Immunological identity of urinary alkaline phosphatase in health and disease.

Urinary alkaline phosphatase has been measured and an investigation has been undertaken involving starch gel electrophoresis after treatment of the enzyme with antisera raised against various human tissues. Urine was obtained from patients with bone and liver disease as well as pregnant women and normal persons. The enzyme level in urine in disease and pregnancy was raised above the normal but considerable overlap occurred. After electrophoresis the most usual finding was a single zone of activity although occasionally very minor zones were also present. Antiserum to kidney had the greatest affinity for the urinary enzyme. Decreasing affinity was found with intestinal and placental antiserum. No affinity was found with liver antiserum.

Alkaline Phosphatase

Surface protein distributions in cells isolated from solid tumours and their metastases.

Methods have been developed which isolate single viable cells from the primary growths of two tumour systems (a lymphosarcoma and a carcinoma) and their secondary deposits. Subsequent comparisons of the surface-membrane structure of pairs of these primary and secondary cells, using lactoperoxidase-catalysed radioiodination coupled with polyacrylamide-gel electrophoresis, suggest that their overall structures are qualitatively very similar. This latter picture is still maintained when the isolated cells are treated with trypsin or incubated in complete medium before radioiodination. Analysis of the incorporated label into defined sections of the electrophoretic patterns revealed small quantitative differences between primary and secondary cells. In particular, slightly reduced incorporation into certain surface components of secondary cell preparations was seen. However, these did not occur for all the animals investigated, and also they did not consistently occur if the isolated cells were incubated in complete medium. The most similar overall change observed for the two tumour systems was a slight reduction in the secondary cells of a 20K mol. wt surface component.

Animals

Increased nuclear size in BHK21 cells after treatment with non-toxic levels of calf thymus histones.

The effect of treatment with either whole calf thymus histones, or individually isolated histones, or polyarginine, or polylysine, on the nuclear size of BHK21 cells has been investigated. Incubation of the cells with mixed histones (12.5--44 microgram/ml) for 1 h considerably increased nuclear size. Increasing the histone concentration and/or the incubation time resulted in a decrease in the effect and could result in no change in nuclear size. Treatment of the cells with polyarginine or polylysine did not affect nuclear size. Experiments with individually isolated histones showed that the nuclear size effect was almost exclusively due to the histone H4. It is argued that the changes observed most likely resulted from interaction of H4 with the nucleus, and could reflect the properties of this particular histone molecule.

Animals

A simple lectin-mediated cell-adhesion method for investigating the cell surface.

A very simple, rapid and reproducible method has been developed for studying the interaction of lectins with the cell surface. This involves determining the number of adherent cells after shaking cell suspensions in Petri dishes which have had a lectin coupled to their surface using 1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluene sulfonate. Using concanavalin A coupled to 60 mm diameter dishes and between 1.5 and 2 x 10(6) tumour cells, this adhesion reached a maximum after 10 min shaking. Maximum cell adhesion also varied according to the particular lectin used. Adhesion was absent or was very low if cells were shaken in untreated dishes, or in dishes coupled to bovine serum albumin, or in the presence of the lectin-specific sugar-competitor. Under conditions of maximum cell adhesion, the binding of two different lymphosarcoma lines to four different lectins was very similar, whereas the binding of a carcinoma line to these lectins was completely different from that observed for the lymphosarcomas.

Animals

Aprotinin induces surface changes in malignant cells in culture: a possible mode of antitumour action.

Treatment of TRES cells with aprotinin (Trasylol) produced marked changes in their surface properties. There was a concentration-dependent increase in the surface charge density of the cells. The agglutinability of the cells was inhibited by the antiproteinase. The cells became less adhesive to concanavalin A-linked plates. The adhesion of treated TRES cells appeared to be mediated by lectin receptors with lower specificity for concanavalin A. These observations and an analysis of the kinetics of adhesion have suggested the possibility that a new class of concanavalin A receptors appears on the surface of aprotinin-treated TRES cells, which might increase immunogenicity of the cells.

Agglutination

Cyclic adenosine 3', 5' monophosphate: a possible indicator of premalignant changes in the large bowel.

Cyclic adenosine 3', 5' monophosphate (cyclic-AMP) has been estimated in mucosal biopsy samples removed from the descending colon and rectum at endoscopy to investigate the possibility of using this substance for monitoring pre-malignant changes in the large bowel. Four groups of patients have been studied: those with normal large bowel and rectal mucosa; those with non-malignant inflammatory bowel disease; those with an adenomatous polyp in the descending colon or sigmoid colon; and those with a rectal adenocarcinoma. No difference was found in the cyclic-AMP content of 'normal' rectal mucosa, 'normal' colonic mucosa, 'diseased' colonic mucosa, carcinomas, and uninvolved mucosa adjacent to the polyps. Less cyclic-AMP was found in the polyps than in adjacent uninvolved mucosa. Conversely, more cyclic-AMP was found in the carcinomas than in adjacent uninvolved mucosa. It is concluded that although cyclic-AMP may be a very useful parameter for delineating the extent of the disease in individual patients, it is not a suitable biochemical marker for the screening of neoplastic changes in the large bowel in the population as a whole.

Colonic Neoplasms

Increased trypsin sensitivity of cell surface macromolecules after malignant transformation.

The effect of treatment with 0.04% (w/v) trypsin (EC 3.4.4.4) for 3 h on the electrophoretic mobility (EPM) of polyoma-virus malignantly transformed BHK21 cells (Py6) and their normal counterparts has been investigated. These particular conditions were chosen because an earlier study had shown that such treatment released material from the Py6 cells which was not obtained from the BHK21 cells. The negative EPM of the Py6 cells at pH 7.5 was greatly increased by this treatment; whereas the EPM of the BHK21 cells remained unchanged. Active enzyme was required to produce the change. No evidence was obtained for cytolysis, cytotoxicity or uptake of the enzyme by the treated Py6 cells. Measurement of the EPM of the Py6 cells at different pH levels before and after trypsin treatment suggested that the enzyme was removing cationic groupings from the cell surface.

Cell Membrane

A method for measurement of low levels of guanosine 3',5'-cylic monophosphate in blood plasma.

A very sensitive method is described for the assay of guanosine 3',5'-cyclic monophosphate (cGMP) based on a competitive protein-binding technique. The procedure is simple to carry out, and does not require the purchase of expensive reagents. The binding-protein used is extracted from rat lungs and the bound and unbound cGMP are separated by ammonium sulphate precipitation. Although prior extraction of the cGMP is required, the time for this extra step is easily compensated for by the shortness of the assay time. The mean cGMP found in the blood plasma of normal young males and females is 9.9 +/- 2.4 (S.D.) nmol/l (range 5.9--14.3 nmol/l) and 9.5 +/- 2.1 (S.D.) nmol/l (range 6.9--12.7 nmol/l), respectively.

Adult

Radioiodination studies of tumour cell-surface proteins after different disaggregation procedures.

The surface of single cells isolated from solid tumours by either a mechanical or an enzymatic method have been compared, using lactoperoxidase-catalyzed radioiodination of the tyrosine-containing proteins. Qualitatively, the patterns of surface labelling were similar, and duplicate experiments indicated that each method of isolation gave reproducible results. Analysis of incorporated label into 4 defined sections of the electrophoretic pattern illustrated quantitative differences. When the cells were isolated mechanically, the incorporation into low-mol.- wt. components was considerably reduced, whereas that into the high-mol.-wt. components was unaffected. Treatment of enzymatically isolated cells with trypsin also reduced incorporation into low-mol.-wt components.

Animals

A long-term evaluation of the CentrifiChem centrifugal microanalyser during routine use in the clinical biochemistry laboratory.

The CentrifiChem centrifugal microanalyser (model 300) has been used routinely in this laboratory for two years. Its reliability for performing total bilirubin, aspartate aminotransferase alkaline phosphatase and gamma-glutamyl transferase analyses during this time is reported, using methods modified in a preliminary study. Accuracy of the analyses has been assessed by comparing the results with those from other analytical systems and by using commercial control sera. Determinations have been made for within-batch, between-day (20 days), and long term (100 weeks) precision. Other aspects evaluated were the range over which methods were linear and the cost of operation.

Alkaline Phosphatase

The effect of surgical operation on venous plasma free amino acids.

The effects of abdominal operation on the antecubital vein plasma amino acid concentration were examined. With four groups, each containing ten patients, factors such as severity of surgical operation, anesthesia, and postoperative nutrition were examined. Immediately after operation there is a fall in the plasma concentrations of most amino acids; the nonessential amino acids (glutamate, proline, glycine, alanine, histidine, and arginine) continue to fall during the 2 postoperative days, whereas the essential amino acids (isoleucine, leucine, phenylalanine, and lysine, with tyrosine) are at higher concentration at 48 hours than immediately after operation. Comparison of a group of patients undergoing an operation of moderate severity (vagotomy and pyloroplasty or cholecystectomy) with those having the more severe procedure of resection of aortic aneurysm with Dacron graft replacement suggests that these general changes in amino acid concentrations do not appear to be due to the severity of the operation, although cystine levels were lower, and phenylalanine levels were higher, in the graft-replacement group. The changes did not appear to be the result of anesthesia. Increasing the postoperative glucose intake was associated with higher plasma alanine and lower methionine levels. It is suggested that, in surgical patients, cystine and tyrosine may become essential amino acids and rises in phenylalanine and methionine indicate transient liver dysfunction. The data give support to the view that a high calorie intake after operation has a beneficial effect.

Abdomen

Human liver alkaline phosphatase purified by affinity chromatography, ultracentrifugation and polyacrylamide-gel electrophoresis.

A method is presented for the preparation of human liver alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1). The method gives a purification factor of 12.5 X 10(3) over the initial aq. butan-1-ol extract, a recovery of 6.0% and a specific activity for the preparation of 1450-1550 units/mg of protein, 1 unit being defined as the amount of enzyme catalysing the hydrolysis of 1mumol of p-nitrophenyl phosphate/min at 35 degrees C in 0.1 M-2-amino-2-methylpropan-1-ol/HCl buffer, pH 10.5, containing 10mM-p-nitrophenyl phosphate. Homogeneity was studied by ultracentrifugation, by immunoelectrophoresis and by polyacrylamide-gel electrophoresis. A single contaminating protein was present which was less than 5% of the total. Ultracentrifugation and equilibrium-gradient-pore electrophoresis techniques indicated a mol.wt. of 156000 and 160000 respectively. Equilibrium-gradient-pore electrophoresis indicated that the alkaline phosphatase molecule is possibly a dimer, comprising two subunits of about 80000 mol.wt. Amino acid analysis proved remarkably similar to that for alkaline phosphatase from other sources, regardless of species.

Alkaline Phosphatase

The separation of plasma lipoproteins using gel electrofocusing and polyacrylamide gradient gel electrophoresis.

Polyacrylamide gel electrofocusing and gradient electrophoresis have been used to separate the lipoproteins in whole plasma and in fractions prepared by sequential flotation in the ultracentrifuge and by precipitation with dextran sulphate and manganous chloride. After the two-dimensional separation, high density lipoproteins appear as a zone showing noticeable heterogeneity with respect to both isoelectric point and molecular weight. Low density lipoproteins are resolved as a compact spot while very low density lipoproteins are visible as a long horizontal streak across the top of the electrophoresis gel. The implications of the technique for the analysis of lipoprotein patterns in pathological plasmas are discussed.

Electrophoresis, Agar Gel