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R Smith

Publications and source records attributed to R Smith.

At least 613 records · Page 34Linked to original sources
PubMed1995

Screening and kinetic analysis of recombinant anti-CEA antibody fragments.

Four different carcinoembryonic antigen (CEA)-binding antibody fragments were prepared using the genes of the variable regions of the T84 epitope-specific antibody 7F7 and phage display techniques. The genes were successfully cloned and expressed in the pCANTAB5 phage display vector to investigate the kinetic binding parameters of each synthesized construct. Single chain fragments, Fab fragments, and two diabodies were purified and compared in their CEA-binding properties with the parent IgG using surface plasmon resonance detection. The on-rates for all these molecules were in the same order of magnitude (about 1 x 10(5) M-1 s-1) whereas major differences were detected in the off-rates. IgG and diabodies had slow off-rates due to bivalent binding, while single chain and Fab fragments dissociated rather fast. We also present a method for the immobilization of large amounts of CEA on CM5 sensorchips. These high density surfaces can be used for observing mass transport limited binding of CEA-specific molecules and are convenient tools for screening and quality control.

Antibody Affinity↗
PubMed1995

Amplification of chromosome band 11q13 and a role for cyclin D1 in human breast cancer.

In this paper we describe how research on the mouse mammary tumor virus model of breast cancer resulted in the identification of an amplified region of DNA on human chromosome 11 band q13. This amplification occurs in approximately 15% of primary breast cancers. Several candidate oncogenes map within the amplicon but by analysing expression of these genes a strong case can be made for a role for cyclin D1 in tumorigenesis. Immunohistochemical staining indicates that cyclin D1 is expressed at elevated levels in around 40% of breast cancers, including those with the 11q13 amplification. The potential function of cyclin D1 as a regulator of early cell division cycle events would be consistent with a role in neoplasia.

Blotting, Southern↗
PubMed1995

1H NMR studies of sarafotoxin SRTb, a nonselective endothelin receptor agonist, and IRL 1620, an ETB receptor-specific agonist.

1H NMR studies on the nonselective endothelin receptor agonist sarafotoxin SRTb have identified a helix between residues Asp 8 and His 16, and a beta-turn involving residues Cys 3 to Met 6; however, the biologically important C-terminal five residues were found to be conformationally variable. The average RMSD, measured for the final 43 refined structures to the average structure over residues 1-16, was 0.78 +/- 0.18 A for the backbone atoms and 1.39 +/- 0.22 A for all atoms. The torsion angles Cys 3 psi/Lys 4 theta, Thr 7 psi/Asp 8 theta and Gln 17 theta were identified as sites of conformational variability. Differences were found between the structures in the bicyclic loop region for SRTb and those published for ET1, another nonselective receptor agonist, which may explain the observed differences in potency of these peptides. The conformation of an ETB receptor-specific agonist, IRL 1620, which lacks the N-terminal seven residues and the two intrachain disulfides, was found by NMR and circular dichroism spectroscopy to be predominantly random coil, despite the fact that its affinity for the ETB receptor almost equals that of ET1. However, close analysis of the NMR results indicated the presence of turn-like structures, or a nascent helix, in the part of the sequence corresponding to the helical region in the parent peptides. These results suggest that the helical conformation may be required for ligand binding to the ETB receptor as well as to the ETA receptor.

Amino Acid Sequence↗
PubMed1995

Breast and ovarian cancer-specific cytotoxic T lymphocytes recognize the same HER2/neu-derived peptide.

The identification of antigenic peptides presented on the tumor cell surface by HLA class I molecules and recognized by tumor-specific cytotoxic T lymphocytes may lead to a peptide vaccine capable of inducing protective cellular immunity. We demonstrate that both HLA-A2-restricted breast and ovarian tumor-specific cytotoxic T lymphocytes recognize shared antigenic peptides. At least one of these peptides is derived from the oncogene product of HER2/neu, which is overexpressed in 30-40% of all breast and ovarian cancers. T cells sensitized against this nine-amino acid sequence demonstrate significant recognition of HLA-A2+, HER2/neu+ tumors. Since 50% of the tumor-cell population is HLA-A2+ and many different tumors express HER2/neu, this peptide may be widely recognized and have many clinical applications.

Antigens, Neoplasm↗
PubMed1995

Genomic organization of the mouse cyclin D1 gene (Cyl-1).

To determine the genomic organization of the mouse cyclin D1 locus (Cyl-1), a series of cosmids and cDNAs were recovered by hybridization with a genomic probe representing the 5' end of the homologous human gene, CCND1. Primer extension indicated that transcripts originate from one of three adjacent nucleotides at a single start site. Two overlapping cDNA clones that essentially accounted for the complete sequence of the larger 4.0-kb Cyl-1 transcript were characterized. A combination of RNase protection and sequencing across intron-exon boundaries established that the gene is organized into five coding exons with a long 3' untranslated region. Repeated attempts to isolate clones corresponding to the minor 3.5-kb RNA were compromised by the presence of an internal poly(A) domain. However, hybridization with specific probes revealed that the minor transcript lacks approximately 800 nucleotides from the 3' end of the major transcript and may be generated by a novel mechanism or by RNA processing.

Amino Acid Sequence↗
PubMed1995

Modeling epidemiologic studies of occupational cohorts for the quantitative assessment of carcinogenic hazards.

Epidemiologic studies of occupational cohorts have played a major role in the quantitative assessment of risks associated with several carcinogenic hazards and are likely to play an increasingly important role in this area. Relatively little attention has been given in either the epidemiologic or the risk assessment literature to the development of appropriate methods for modeling epidemiologic data for quantitative risk assessment (QRA). The purpose of this paper is to review currently available methods for modeling epidemiologic data for risk assessment. The focus of this paper is on methods for use with retrospective cohort mortality studies of occupational groups for estimating cancer risk, since these are the data most commonly used when epidemiologic information is used for QRA. Both empirical (e.g., Poisson regression and Cox proportionate hazards model) and biologic (e.g., two-stage models) models are considered. Analyses of a study of lung cancer among workers exposed to cadmium are used to illustrate these modeling methods. Based on this example it is demonstrated that the selection of a particular model may have a large influence on the resulting estimates of risk.

Cadmium↗
PubMed1995

Structural evaluation of human and sheep bone and comparison with synthetic hydroxyapatite by FT-Raman spectroscopy.

The composition of whole human and sheep cortical bone tissue, and of a synthetic hydroxyapatite (P120), were compared using Fourier transform Raman (FT-Raman) spectroscopy. Deproteination procedures to remove the bulk of the collagen present in bone tissue allowed isolation of the mineral phase. A comparison of the spectra obtained from both whole and deproteinated bone with those of synthetic hydroxyapatite showed direct correlation only in the region of 952 cm-1 (symmetric P-O mode). In contrast, human and sheep bone were very closely matched in both, the organic and inorganic structures. The results demonstrate that deproteination of bone is not a necessary precursor to obtain spectral information.

Animals↗
PubMed1995

Computer-assisted mapping of infarcted and viable regions of gross cardiac sections following experimental myocardial infarction.

We have developed a computer-assisted method that creates digital maps of the viable and infarcted regions of tetrazolium-stained gross cardiac sections. Here we describe and test the method, using a canine occlusion-reperfusion infarction model. Quantitative image analysis showed that the method accurately recorded differences between infarcted and viable regions at a spatial resolution of 5-20 pixels per mm2. Microscopic analysis of tissue samples taken from the sections showed that the maps were accurate. In 15 of 15 cases the histology of the samples matched that predicted by the maps. A comparison of infarct area measurements derived from the maps showed that the method was reproducible. The average intraoperator standard deviation was +/- 8% of a slice's infarcted area and the average interoperator standard deviation was +/- 20%. We conclude that the method creates accurate, detailed, and reproducible maps of the infarcted and viable regions of tetrazolium-stained cardiac sections.

Animals↗