Search PubMed⌕ Search

Biomedical subjects

M A Kerr

Publications and source records attributed to M A Kerr.

At least 55 records · Page 3Linked to original sources
PubMed1994

Classification of CEA-related positivity in primary and metastatic malignant melanoma.

Using a panel of antibodies to carcinoembryonic antigen (CEA), in paraffin-processed biopsy material patchy, predominantly membranous positivity was seen on tumour cells in 70 per cent of cases of superficial spreading melanoma, 60 per cent of nodular melanomas, and 75 per cent of secondary deposits studied with unabsorbed polyclonal anti-CEA only. No staining was seen using monoclonal anti-CEAs. Localization of CEA to the cell membrane was confirmed with confocal microscopy. Immunoblotting of fresh frozen material detected CEA of around 180 kD in both primary and metastatic melanomas migrating with an apparent molecular weight of between 150 and 200 kD, indicating variable glycosylation of the protein. Recognition of an adhesive role for CEA with roles in immunolocalization and immunotherapy emphasizes the importance of more precise classification of CEA-related positivity in human tumours.

Carcinoembryonic Antigen↗
PubMed1994

Classification and localisation of carcinoembryonic antigen (CEA) related antigen expression in normal oesophageal squamous mucosa and squamous carcinoma.

Using a panel of carcinoembryonic antigen (CEA) related antibodies in normal oesophageal squamous mucosa CEA expression is present on suprabasal squames localised to the cell membrane. Immunoblotting shows that this positivity is predominantly due to a glycoprotein of around 180 kDa representing CEA itself. Positivity in squamous carcinomas is confined to cells in foci of squamous differentiation. A shift from membranous localisation to predominant cytoplasmic overexpression is shown between normal and malignant squames using confocal microscopy. The recognition of an adhesive role for CEA and a role in enhancing distant metastases in those tumours expressing CEA highlights the importance of recording CEA expression and changes in subcellular distribution between normal and malignant tissues; CEA expression in oesophageal squamous mucosa has not been well recognised previously and changes in expression may prove of great significance in the spread and dissemination of squamous carcinoma.

Carcinoembryonic Antigen↗
PubMed1993

Neutrophil NCA-160 (CD66) is the major protein carrier of selectin binding carbohydrate groups LewisX and sialyl lewisX.

The neutrophil surface carbohydrate groups LewisX and sialyl LewisX have previously been shown to interact with the vascular selectins E- and P-selectin. However, the proteins expressing these carbohydrate groups have not been fully characterised. We show that these carbohydrate groups, and the structurally related Lewisa group, are all carried predominantly on a sub-population of the carcinoembryonic antigen (CEA) related NCA-160, which is also recognised by CD66 antibodies. We demonstrate that the related NCA-160, NCA-90/95 and CD67 all undergo an increase in surface expression in response to fMLP stimulation, and that this increase is greater than that observed for the sialyl and non-sialyl LewisX carbohydrate groups. These data suggest that the expression of differently glycosylated forms of NCA is independently regulated.

Antigens, CD↗
PubMed1993

Design, synthesis and biological activity of protaxols.

Taxol is a product isolated from the Pacific yew tree (Taxus brevifolia) and is a potent microtubule-stabilizing agent which has recently been approved for treatment of otherwise intractable ovarian cancer. Despite taxol's therapeutic promise, its aqueous insolubility (< 0.004 mg ml-1) hampers its clinical application. Here we report the design, synthesis and biological activity of a series of taxol-releasing compounds (protaxols) with improved pharmacological properties. These prodrugs were designed to increase their aqueous solubility and allow for taxol release under basic or physiological conditions. We demonstrate the stability of these prodrugs at pH < or = 7 and their ability to release taxol in a basic medium. Taxol-like microtubule-stabilizing activity appears after the release of taxol. In vitro these prodrugs have cytotoxic properties against tumour cell lines comparable to those of taxol; moreover, human plasma catalyses the release of active taxol. These protaxols have greater potential as anticancer agents than the parent compounds taxol and taxotere (Fig. 1a).

Animals↗
PubMed1993

The cleavage of immunoglobulin G in vitro and in vivo by a proteinase secreted by the urinary tract pathogen Proteus mirabilis.

Eighteen different strains of Proteus mirabilis were all shown to produce an EDTA-sensitive proteinase of c. 50 kDa that cleaved the heavy chain, but not the light chain, of IgG. Digestion of pure IgG with small amounts of pure P. mirabilis proteinase generated Fabc'2 and Fab'2 fragments; greater amounts generated Fab and Fc fragments that were comparable in size to those generated by pepsin and papain, respectively. Incubation of neutrophils with IgG digested with P. mirabilis proteinase or papain resulted in a marked decrease in the respiratory burst activity of the neutrophils that coincided with cleavage of the IgG into Fab and Fc fragments. Analysis of urine from patients with P. mirabilis urinary tract infection revealed in many the presence of Fab and Fc fragments of IgG indistinguishable in size from those generated by P. mirabilis proteinase. These results indicate that, in P. mirabilis urinary tract infections, the proteinase is secreted and cleaves IgG to fragments that have defective immune effector functions, thereby limiting the effectiveness of the immune response.

Adolescent↗
PubMed1992

Stimulation of neutrophil adhesion by antibodies recognizing CD15 (Le(X)) and CD15-expressing carcinoembryonic antigen-related glycoprotein NCA-160.

The carbohydrate antigen, CD15 (Le(X)), and its sialylated derivative have recently been shown to be involved in the binding of neutrophils to the endothelial lectins, E-selectin and P-selectin. Neutrophil NCA-160, a carcinoembryonic antigen (CEA)-related glycoprotein, is the major carrier of CD15, which is also expressed on the common beta 2 chain of leucocyte integrins. Rabbit IgG antibodies directed against CEA, which cross-react with neutrophil NCAs, increase the adhesion of neutrophils to plastic. This effect is also observed with F(ab')2 and Fab antibody fragments and a monoclonal antibody (mAb) recognizing the same antigen. Anti-CD15 mAbs inhibit adhesion at higher concentrations, but augment adhesion at lower concentrations. Anti-CEA and anti-CD15 antibodies cause the homotypic adhesion of neutrophils demonstrable by light microscopy and flow cytometry. Anti-(integrin beta 2 chain) mAbs inhibit both adhesion to plastic and homotypic adhesion. These results suggest that binding of ligand to NCA-160 is able to trigger neutrophil adhesion events which have been shown to be integrin mediated. Anti-CD15 mAbs do not, however, induce a respiratory burst from neutrophils.

Antibodies↗
PubMed1992

Mammary development and milk secretion in transgenic mice expressing the sheep beta-lactoglobulin gene.

Mammary development and milk secretion were studied in transgenic mice which exhibited mammary tissue-specific expression of the sheep beta-lactoglobulin gene, and secreted significant quantities of the foreign protein in milk. Mammary development was unaffected by transgenesis. Tissue DNA content and the activities of several key enzyme markers of cell differentiation were similar in transgenic mice and non-transgenic controls. Milk yield, whether estimated by pup weight gain or measured by a 3H2O-dilution method, was unchanged by foreign gene expression. Gross milk composition, including milk protein concentration, was also similar in transgenic and non-transgenic animals, even though beta-lactoglobulin accounted for 29% of total milk protein. Therefore the foreign gene product was synthesized at the expense of endogenous milk proteins. However, transgenic mammary tissue in vitro exhibited a significantly higher rate of total protein synthesis than did control tissue. This suggested that a factor limiting milk protein synthesis or secretion in transgenic mice in vivo may have been removed by short-term explant culture of mammary tissue. The results emphasize that the use of transgenesis for manipulating milk composition may depend not only on high-level mammary-specific expression of the foreign gene, but also on the biosynthetic capacity of the mammary gland itself.

Aging↗
PubMed1992

The use of spent renal dialysis membranes for the isolation of large numbers of human neutrophils for biochemical studies. Application to purification of the myeloid IgA receptor (Fc alpha R).

Human neutrophils (PMN) can be eluted from spent Cuprophan renal dialysis membranes in large numbers (10(9)-10(10) per dialyser cartridge) and in relatively high purity by washing the membranes with 0.35 M NaCl. This offers the possibility of isolating relatively large amounts (10(-4)-10(-3) g) of minor PMN proteins such as those expressed on the cell surface. Here the technique is applied to the purification of the neutrophil IgA receptor (Fc alpha R). Affinity chromatography on IgA-Sepharose of NP-40 extracts of 125I-labelled PMN isolated from fresh venous blood routinely gave a receptor preparation showing one diffuse band, Mr 50-70 kDa, upon analysis by SDS-PAGE and autoradiography. When the same method was used with larger numbers of unlabelled PMN from fresh venous blood or renal dialysis membranes a preparation was obtained which gave multiple bands upon analysis by SDS-PAGE silver stained gels due to contamination of the receptor with cytoplasmic proteins which bound non-specifically to the IgA-Sepharose. Most of these contaminants could be removed by chromatography of the IgA-Sepharose eluates on wheat germ agglutinin-Sepharose.

Antigens, CD↗
PubMed1992

Membranous expression of carcinoembryonic antigen (CEA) in the normal cervical squamous mucosa.

Using a panel of polyclonal and monoclonal antibodies to CEA-related antigens in paraffin-processed cervical biopsies, CEA and NCA expression has been demonstrated on the cell membrane of normal mature cervical squames. Cytofluorimetry of disaggregated cervical squames confirmed membranous expression and immunogold labelling of ultrathin cryostat sections localized CEA to the glycocalyx and to within cytoplasmic membrane-bound vesicles of mature squames. Immunoblotting of cervical tissue showed that most of the CEA reactivity was expressed as a glycoprotein of molecular weight around 180 k, probably CEA itself. Localization of the CEA to the cell membrane of mature cervical squames suggests a key role for these antigens in maintaining the integrity of the squamous mucosa, through the putative function of an adhesion molecule.

Carcinoembryonic Antigen↗
PubMed1992

Dissociation of primary antigen-antibody bonds is essential for complement mediated solubilization of immune precipitates.

The role of dissociation of primary antigen-antibody bonds in the solubilization of immune complexes (IC) has been investigated using photo-affinity crosslinked IC comprising NAP15-BSA and murine monoclonal anti-DNP antibodies. Non-covalently linked IC were solubilized rapidly when incubated with normal human serum (NHS), whereas covalently-linked IC were solubilized poorly or not at all. The rate and extent of complement activation produced by incubating covalently-linked and non-covalently linked IC with NHS was similar as assessed by the production of the C1s:C1-inhibitor, C3:properdin and C5b-9 complexes and the anaphylatoxins C4a and C3a. Thus, the inability of serum to solubilize photo-affinity crosslinked IC must be due to failure of dissociation of primary antigen-antibody bonds.

Animals↗
PubMed1992

Separate and additive stimulation of bovine milk yield by the local and systemic galactopoietic stimuli of frequent milking and growth hormone.

Lactating heifers were treated for 4 weeks with recombinant bovine growth hormone (bGH, n = 9) or were untreated (n = 9). In addition, two mammary glands of each heifer were milked four times daily rather than the normal twice daily for the same 4 weeks, and for the following 2 weeks. Over the 4 weeks, milk yield was increased 12.8% by bGH, 14.0% by frequent milking and 28.5% by the combined treatment. The effect of bGH as administered here was slower in onset than that of frequency, but eventually produced a higher peak yield. ANOVA revealed significant effects of each stimulus independently and an additive, but not synergic effect of the combined treatment. The effect of the combined treatment tended to persist beyond the end of treatment; most of this response was related to the milking frequency component rather than the bGH. Mammary differentiation was assessed in biopsies of mammary tissue obtained prior to and at the end of treatment. Mammary enzyme activities (expressed on a per cell basis) indicated minimal differentiative response to either treatment, but synthesis rates for lactose and casein determined in vitro were increased by bGH treatment. Histological examination revealed a stimulatory effect of milking frequency on epithelial cell size. The results indicate that these two galactopoietic stimuli operate through independent mechanisms, and neither stimulus alone is sufficient to maximize milk yield in dairy heifers.

Acetyl-CoA Carboxylase↗
PubMed1992

Effect of stage of lactation and milk accumulation on mammary cell differentiation in lactating bats.

Mammary cell differentiation was measured in lactating pipistrelle bats (Pipistrellus pipistrellus) by assay of key enzyme activities, and by determination of protein and lactose synthesis rates in short-term tissue cultures. By these criteria, mammary cell differentiation did not change significantly with stage of lactation, but depended on the extent to which the gland was filled with milk. Key enzyme activities and in vitro synthesis rates were significantly higher in glands suckled immediately before tissue collection, compared with contralateral glands that were engorged with milk. This indicates that mammary cell differentiation in the lactating bat is regulated locally within each gland by a mechanism sensitive to milk accumulation, to the extent that, unlike other species, this obscures any underlying effect of stage of lactation.

Animals↗
PubMed1992

Proteinases of Proteus spp.: purification, properties, and detection in urine of infected patients.

The proteinases secreted by pathogenic strains of Proteus mirabilis, P. vulgaris biotype 2, P. vulgaris biotype 3, and P. penneri were purified with almost 100% recovery by affinity chromatography on phenyl-Sepharose followed by anion-exchange chromatography. The proteinase purified from the urinary tract pathogen P. mirabilis, which we had previously shown to degrade immunoglobulins A and G, appeared as a composite of a single band and a double band (53 and 50 kDa, respectively) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The other Proteus proteinases had similar patterns but slightly different mobilities. In each case all proteinase activity in culture supernatants was demonstrated by gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be associated with only the triple-band complex; all three bands were proteolytically active. The P. mirabilis proteinase was resistant to inhibitors of both serine and thiol proteinases but strongly inhibited by metal chelators, although it was not affected by phosphoramidon, an inhibitor of the thermolysin group of bacterial metalloproteinases. Active proteinase was detected in urine samples from P. mirabilis-infected patients; this is consistent with our detection of immunoglobulin A fragments of a size suggestive of P. mirabilis proteinase activity.

Chromatography, Affinity↗
PubMed1991

Purification and characterization of human immunoglobulin IgA1 and IgA2 isotypes from serum.

A method is described for the simultaneous purification of IgA1 and IgA2 from human serum. Ammonium sulphate precipitation, gel filtration and ion-exchange chromatography on DEAE-Sephacel yielded a partially purified IgA preparation which was separated quantitatively into IgA1 and IgA2 by affinity chromatography on jacalin-Sepharose. The IgA1 which bound to the jacalin was eluted with 0.8 M D-galactose. The IgA1 preparation was apparently homogeneous by SDS-PAGE but contained a trace of C1-inhibitor and a second protein detected by immunoelectrophoresis. The IgA2 which did not bind to the jacalin was purified to apparent homogeneity by chromatography on columns of Protein G-Sepharose, Fastflow-S Sepharose and Superose 6. Typical yields were 95% and 58% for IgA1 and IgA2 respectively or 253 mg and 24 mg per 100 ml serum. The IgA1 and IgA2 were characterised by their reactivity with isotype specific monoclonal antibodies and sensitivity to bacterial proteinases. The IgA2 preparation apparently contained both allotypes, IgA2m(1) and IgA2m(2).

Chromatography, Gel↗
PubMed1991

The production and activity in vivo of Proteus mirabilis IgA protease in infections of the urinary tract.

Immunoblotting of urine from 21 patients of both sexes and of wide age range who had a Proteus mirabilis urinary tract infection (UTI) showed that 14 (64%) specimens contained immunoglobulin A (IgA). In nine (64%) of these the IgA heavy chain had been degraded to fragments of a size identical to those formed when purified IgA was degraded by pure P. mirabilis protease. Urine from patients with clinical evidence of upper UTI contained fragmented IgA and in some of these urine samples P. mirabilis protease activity was detectable. Urine infected with a non-proteolytic strain contained only intact IgA. It is concluded that P. mirabilis IgA protease is produced and is active during infections of the urinary tract.

Adolescent↗
PubMed1991

Intracellular degradation of newly synthesized casein in perfused rat mammary gland.

Degradation of newly synthesized casein in rat mammary gland perfused in situ was measured by a pulse-chase method using [3H]proline. Casein degradation during secretion was observed in the absence of prolactin, but not with prolactin present. Partial inhibition by chloroquine showed that hormone-dependent degradation occurred intracellularly by a lysosomal mechanism. The study indicates that this post-translational mechanism is a physiological regulator of net casein secretion.

Adenosine Triphosphate↗
PubMed1991

The measurement of respiratory burst induced in polymorphonuclear neutrophils by IgA and IgG anti-gliadin antibodies isolated from coeliac serum.

The properties of IgA and IgG anti-gliadin antibodies from the serum of patients with coeliac disease have been compared. The antibodies were quantified by ELISA using microtitre plates coated with crude gliadin fractions. Their specificity was confirmed by immunoblotting. Heat-treated sera containing IgA antibodies stimulated chemiluminescence when added together with neutrophils to microtitre plates coated with crude gliadin. Sera containing only IgG antibodies were less efficient. When IgA and IgG were purified from a serum containing both classes of anti-gliadin antibodies, each of the preparations was able to stimulate neutrophil chemiluminescence in plates coated with gliadin. Although the yield of anti-gliadin antibody determined by ELISA was high, the ability of the purified immunoglobulins to stimulate neutrophil chemiluminescence was much less than that of the unfractionated serum. This loss of activity was shown to be due to the ability of each class of antibody to potentiate the activity of the other in the whole serum.

Adolescent↗