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A Murray

Publications and source records attributed to A Murray.

At least 181 records · Page 10Linked to original sources
PubMed1998

Summary report on the ISOBM TD-4 Workshop: analysis of 56 monoclonal antibodies against the MUC1 mucin. San Diego, Calif., November 17-23, 1996.

Sixteen research groups participated in the ISOBM TD-4 Workshop in which the reactivity and specificity of 56 monoclonal antibodies against the MUC1 mucin was investigated using a diverse panel of target antigens and MUC1 mucin-related synthetic peptides and glycopeptides. The majority of antibodies (34/56) defined epitopes located within the 20-amino acid tandem repeat sequence of the MUC1 mucin protein core. Of the remaining 22 antibodies, there was evidence for the involvement of carbohydrate residues in the epitopes for 16 antibodies. There was no obvious relationship between the type of immunogen and the specificity of each antibody. Synthetic peptides and glycopeptides were analyzed for their reactivity with each antibody either by assay of direct binding (e.g. by ELISA or BiaCore) or by determining the capacity of synthetic ligands to inhibit antibody binding interactions. There was good concordance between the research groups in identifying antibodies reactive with peptide epitopes within the MUC1 protein core. Epitope mapping tests were performed using the Pepscan analysis for antibody reactivity against overlapping synthetic peptides, and results were largely consistent between research groups. The dominant feature of epitopes within the MUC1 protein core was the presence, in full or part, of the hydrophilic sequence of PDTRAPAP. Carbohydrate epitopes were less easily characterized and the most useful reagents in this respect were defined oligosaccharides, rather than purified mucin preparations enriched in particular carbohydrate moieties. It was evident that carbohydrate residues were involved in many epitopes, by regulating epitope accessibility or masking determinants, or by stabilizing preferred conformations of peptide epitopes within the MUC1 protein core. Overall, the studies, highlight concordance between groups rather than exposing inconsistencies which gives added confidence to the results of analyses of the specificity of antimucin monoclonal antibodies.

Amino Acid Sequence↗
PubMed1998

Epitope mapping of anti-MUC1 mucin protein core monoclonal antibodies.

A panel of 56 murine monoclonal antibodies against the MUC1 mucin was analysed for reactivity against purified MUC1 mucin and synthetic MUC1 mucin protein core-related peptides. In an ELISA assay, with purified normal urinary mucin as the target antigen, 48/56 (86%) of antibodies showed positive reactivity. A smaller proportion of antibodies, 31/56 (55%), reacted with a bovine serum albumin conjugate containing the synthetic peptide, APDTRPAPG. This peptide represents the hydrophilic region of the tandem repeat sequence of the MUC1 mucin core. These 31 anti-mucin core antibodies were then evaluated for reactivity with a set of overlapping heptamers with sequences based upon that of the 20-amino acid tandem repeat of the MUC1 protein core. For this purpose, the 'Pepscan' procedure was employed to detect antibody binding to peptides tethered at their carboxy termini to the polypropylene pins. Synthetic peptides binding to these antibodies were identified for 29 of the samples provided and epitopes or 'minimum binding units' were deduced for these. Most antibodies bound to epitopes of 4 amino acid residues with the sequence RPAP occurring most frequently. The results confirm that the short hydrophilic region PDTRPAP in the MUC1 mucin core is immunodominant in the induction of antibodies to the MUC1 mucin since half of the antibodies submitted reacted with this peptide.

Amino Acid Sequence↗
PubMed1998

Interobserver variability in recognizing arousal in respiratory sleep disorders.

Daytime sleepiness is a common consequence of repeated arousal in obstructive sleep apnea (OSA). Arousal indices are sometimes used to make decisions on treatment, but there is no evidence that arousals are detected similarly even by experienced observers. Using the American Sleep Disorders Association (ASDA) definition of arousal in terms of the accompanying electroencephalogram (EEG) changes, we have quantified interobserver agreement for arousal scoring and identified factors affecting it. Ten patients with suspected OSA were studied; three representative EEG events during each of light, slow-wave, and rapid-eye-movement (REM) sleep were extracted from each record (90 events total) and evaluated by experts in 14 sleep laboratories. Observers differed (ANOVA, p < 0.001) in the number of events scored as arousal (totals ranged from 23 to 53 of the 90 events). Overall agreement was moderate (kappa = 0.47), but it was best for events during slow-wave sleep, moderate for REM, and poor for light sleep (kappa = 0.60, 0.52, and 0.28, respectively). Agreement was unrelated to arousal duration. We conclude that the ASDA definition of arousal is only moderately repeatable. Account should be taken of this variability when results from different centers are compared.

Adult↗
PubMed1998

Recombinant heregulin-Pseudomonas exotoxin fusion proteins: interactions with the heregulin receptors and antitumor activity in vivo.

Growth factor receptors provide unique opportunities for development of targeted anticancer therapy. Members of the type I receptor tyrosine kinase family, including epidermal growth factor (EGF) receptor (EGFR) and ErbB-2/neu, are often overexpressed in various human cancer cells, including breast. Recently, it has been shown that both ErbB-3 and ErbB-4 are receptors for heregulin (HRG)/Neu differentiation factor. Eight chimeric toxins composed of the extracellular and EGF-like domains of four different HRG isoforms and truncated Pseudomonas exotoxin (PE38KDEL) were constructed. The fusion proteins exhibited activity similar to the native HRG in inducing ErbB receptors phosphorylation. The EGF-like domain of HRG13 and HRGbeta2 fused to PE38KDEL showed the highest cytotoxic activity, with a IC50 of < or = 0.001 ng/ml. The alpha isoforms that were fused to PE38KDEL were 100-fold less active than the beta isoforms. The HRG-Pseudomonas exotoxin (PE) toxins show extremely high activity against cells expressing ErbB-4 receptor, alone or together with other members of the ErbB receptor family. Cells that do not express ErbB-4 but express ErbB-3 receptor, together with the ErbB-2 or EGFR, exhibited moderate sensitivity to HRG-PE toxins. HRG-PE toxins have little or no activity against cells expressing EGFR, ErbB-2, or ErbB-3 alone. More than an 80% tumor regression was achieved by intratumor injection of 1 microg of fusion proteins per day for 5 days. Continuous i.p. administration of EGF-like domain of HRGbeta1-PE38KDEL for 7 days via a miniosmotic pump at a dose of 40 microg/kg/day inhibited the growth of ErbB-4 receptor positive but not ErbB-4 receptor negative cell lines in athymic nude mice. We conclude that there is therapeutic potential of HRG-PE toxins in the therapy of cancers overexpressing the ErbB-4 or ErbB-2 plus ErbB-3 receptors.

Animals↗
PubMed1998

Automated feature extraction and identification of colon carcinoma.

OBJECTIVE: To assess an automated algorithm, developed for the classification of normal and cancerous colonic mucosa, using geometric analysis of features and texture analysis. STUDY DESIGN: Twenty-one images were analyzed, 10 from normal and 11 from cancerous mucosa. The classification was based on a regularity index dependent on shape, object orientation for establishing parallelism and five texture features derived using the co-occurrence image analysis method. RESULTS: Geometric analysis yielded an overall classification accuracy of 80%. The corresponding sensitivity and specificity were 94% and 64%, respectively. Using texture analysis, the overall classification accuracy was 90%, with a sensitivity and specificity of 82% and 100%, respectively. CONCLUSION: This initial study demonstrated that geometric and texture analysis techniques show promise for automated analysis of colon cancer.

Algorithms↗
PubMed1997

Purification of monoclonal antibodies by epitope and mimotope affinity chromatography.

Murine monoclonal antibodies raised against the carcinoma-associated MUC1 mucin have applications in the diagnosis and therapy of human cancer. Many of these antibodies define linear epitopes of three, four or five amino acids within an immunodominant region of the MUC1 protein core. Various synthetic peptides which incorporated this region were prepared and covalently linked to agarose beads for use as affinity matrices. An unrelated peptide was identified as a mimotope for one of the anti-MUC1 antibodies using phage display technologies and this was also evaluated as a potential ligand in an affinity matrix. Epitope affinity chromatographic purification of an anti-MUC1 antibody was performed using hybridoma tissue culture supernatants as sample. Following sample application and column washing, antibody was desorbed from the matrix by gradient elution with increasing concentrations of NaSCN. The procedure has proved efficient for the purification of anti-MUC1 antibodies and the concentration of NaSCN required for antibody desorption gives a measure of the relative binding affinity of the antibody for the peptide epitope matrix so that separation strategies may be optimised.

Amino Acid Sequence↗
PubMed1997

Identification and differentiation of mycobacteria using the PAN promoter sequence from Mycobacterium paratuberculosis as a DNA probe.

A 165 bp DNA fragment containing the PAN promoter from Mycobacterium paratuberculosis was used as a probe in Southern blots to detect the presence of related sequences in other species of mycobacteria. Among the species tested homologous sequences appeared to be present in representative pathogens belonging to the Mycobacterium tuberculosis complex, the MAIS complex, Mycobacterium kansasii and also the non-pathogenic vaccine strain Mycobacterium bovis BCG. In addition, the probe could differentiate between these species on the basis of a restriction fragment length polymorphism (RFLP). No hybridization was observed with DNA extracted from a selected group of other slow-growing and fast-growing mycobacteria nor from a selection of other bacterial pathogens. It appears that the PAN sequence is identifying genomic regions common to the major pathogenic groups of mycobacteria.

Base Sequence↗
PubMed1997

Optical assessment of recovery of tissue blood supply after removal of externally applied pressure.

The authors use photoelectric plethysmography to determine the external occlusion pressure for blood vessels in human tissue in vivo. Three wavelengths are employed; 950 nm (infra-red), 640 nm (red) and 583 nm (yellow). Each probe is applied in turn to one finger of each subject. Pressure is applied, using a neonatal blood pressure cuff, to the finger via the probe. This pressure is increased linearly to 20 kPa (150 mmHg) over 15 s and then decreased linearly to zero over 15 s. The pressure at which perfusion returns is obtained for four repeat measurements at each wavelength. The mean (+/-standard deviation) occlusion pressures for all 13 subjects investigated are 7.1 (+/-1.9) kPa for infra-red, 6.3 (+/-1.7) kPa for red and 5.8 (+/-1.8) kPa for yellow. The pressure is 0.79 (+/-0.83) kPa lower for red compared with infra-red (P < 0.01), 0.54 (+/-0.60) kPa lower for yellow compared with red (P < 0.002) and 1.3 (+/-1.0) kPa lower for yellow compared with infra-red (P < 0.005). The reduced penetration of shorter optical wavelengths can be used to detect the lower occlusion pressures of the smaller blood vessels nearer the skin surface.

Adult↗
PubMed1997

Detection of Mycobacterium avium subsp. paratuberculosis in ovine tissues and blood by the polymerase chain reaction.

A direct polymerase chain reaction (PCR) test was applied to DNA extracted from blood, liver, ileocecal lymph node and ileum from twelve ewes in poor condition with histologically confirmed paratuberculosis and ten clinically normal sheep which had no evidence of paratuberculosis. The assay was compared with four serological tests: complement fixation test (CFT), gel diffusion test (AGID) and two enzyme-linked immunosorbent assays (ELISA). The PCR detection rate of Mycobacterium avium subsp. paratuberculosis, when results of single tests were interpreted in duplicate, was 72% for ileocecal lymph node, 90% for liver, and 100% for ileum in sheep with confirmed paratuberculosis. A single PCR test detected the target DNA in 66% of 0.5 ml blood samples. Sensitivities of serological tests compared with histological diagnosis were: 33% for CFT, 66% for AGID, 75% for the Central Animal Health Laboratory (CAHL) ELISA, and 83% for the 'modified' Commonwealth Serum Laboratories (CSL) ELISA. The PCR assay gave no positive reaction in samples collected from 10 sheep considered to be free of paratuberculosis. Similarly, all four serological tests were also 100% specific. The results raise some hope for the development of a PCR-based test using liver biopsy specimens, or possibly blood, in the diagnosis of paratuberculosis in sheep.

Animals↗
PubMed1997

Patient comfort during cataract surgery with modified topical and peribulbar anesthesia.

PURPOSE: To evaluate patient comfort during cataract surgery performed using modified topical and peribulbar anesthesia. SETTING: Sunderland Eye Infirmary Cataract Treatment Centre, Sunderland, United Kingdom. METHODS: In this prospective study, 50 consecutive patients scheduled for cataract surgery with local anesthesia were randomly assigned to receive topical bupivacaine hydrochloride plus 0.1 mL subconjunctival lignocaine or standard peribulbar anesthesia. All surgery was performed by one surgeon using a scleral pocket bimanual phacoemulsification technique. Patients were asked to grade the pain experienced during administration of the anesthetic and during cataract surgery using a visual analog scale. RESULTS: No statistically significant difference in patient comfort was demonstrated between the two groups during cataract surgery (P < .4, Wilcoxon rank-sum test), and all patients were satisfied with the anesthesia. However, administration of topical bupivacaine was significantly less painful than peribulbar injections (P < .001, Wilcoxon rank-sum test). In eight patients in the topical anesthesia group, the cataract in the fellow eye was removed using peribulbar anesthesia; seven of these patients stated a preference for the modified topical method, while one patient thought there was no difference between the two methods. There was no difference between the groups in surgical complications, but 24% of the peribulbar group had a minor subconjunctival hemorrhage at the needle entry site. CONCLUSION: The modified topical technique provided satisfactory patient comfort during cataract surgery; it was comparable to the comfort achieved using peribulbar injections. The speed and ease of administering topical anesthesia coupled with the rapid visual recovery after surgery makes this method a suitable and safe choice for day-case phacoemulsification cataract surgery.

Administration, Topical↗
PubMed1997

Peritonsillar haematoma.

We present one patient who was admitted with a peritonsillar haematoma as a complication of acute tonsillitis and concomitant warfarin therapy for prosthetic aortic valves. While tonsillar haemorrhage as a result of acute tonsillitis has been well described, no cases of isolated haematoma formation have been documented, nor has it been recognized previously as a complication of long-term anticoagulant therapy. We discuss establishing the diagnosis, the likely aetiology and implications of this complication.

Acute Disease↗
PubMed1997

In vitro responses to noradrenaline of small intestine taken from normal and grass sickness-affected horses.

Small intestine was taken from the caudal flexure of the duodenum and the terminal ileum proximal to the ileocaecal fold of 25 horses, 9 with acute grass sickness (AGS), 12 with subacute grass sickness (SAGS) and 12 with chronic grass sickness (CGS). The motility in the samples was measured isometrically either within 1 h of death or after storage for 24 h at 4 degree C. In control tissue, noradrenaline produced contractions of muscle strips which did not involve a muscarinic cholinergic mechanism and which were unaffected by the alpha 1 antagonist prazosin but were blocked by the alpha 2 antagonist yohimbine. Pretreatment with the alpha antagonist phentolamine prevented the contractile response to noradrenaline and the background contractions either continued at a reduced rate and amplitude or were abolished after a few minutes. Thus, following alpha blockade, noradrenaline reduced the background contraction rate by an effect on inhibitory beta adrenoceptors. The rate of background contractions in duodenal preparations was significantly greater than that in control ideal preparations. Although cold storage for 24 h caused a reduction in the background contraction rates of the control preparations, there was no effect on the contractile responses to noradrenaline, the associated pharmacology being similar to that of fresh tissue. This suggests that noradrenaline-evoked contraction was not dependent on enteric neural elements. The response to noradrenaline by grass sickness-affected tissue was generally similar to that of tissue from control horses, with an immediate contraction which was alpha 2 sensitive. The contractile response to noradrenaline after propranolol was significantly reduced in the CGS group and there were significant differences between the AGS, CGS and control groups. There was a significant difference between the ileal preparations from the control and SAGS groups in their response to noradrenaline following pretreatment with propranolol.

Adrenergic alpha-Agonists↗
PubMed1997

Light microscopy of the enteric nervous system of horses with or without equine dysautonomia (grass sickness): its correlation with the motor effects of physostigmine.

Light microscopy was undertaken on sections from the caudal flexure of the duodenum and the terminal ileum proximal to the ileocaecal fold in 5 control horses, 5 horses with acute grass sickness (AGS), and 5 horses with chronic grass sickness (CGS). With the exception of the ileal submucous plexus of the CGS group, the AGS group had the lowest number of neurons as measured using a subjective scoring scheme. The proportion of abnormal neurons in the AGS group was similar in both plexuses and both regions, whereas the values for the CGS group were much higher in the duodenal region than in the ileal region. The motility of tissue adjacent to that used for histology was measured isometrically in vitro. The increase in the rate of contractions following exposure to physostigmine was greatest for the AGS group, both from the duodenal and from the ileal region. The latency was longest for the AGS group, suggesting that the material from this group had the least number of active cholinergic neurons. The studies with physostigmine thus indicated that the most severe functional damage occurred in cases of AGS. These findings confirm that extensive damage occurs in the enteric neurons in equine grass sickness. There was good correlation between the functional cholinergic responses and the extent of neuronal degeneration.

Acute Disease↗
PubMed1997

Human breast cancer cell metastasis to long bone and soft organs of nude mice: a quantitative assay.

Bone is a common metastatic site in human breast cancer (HBC). Since bone metastasis occurs very rarely from current spontaneous or experimental metastasis models of HBC cells in nude mice, an arterial seeding model involving the direct injection of the cells into the left ventricle has been developed to better understand the mechanisms involved in this process. We present here a sensitive polymerase chain reaction (PCR) method to detect and quantitate bone and soft organ metastasis in nude mice which have been intracardially inoculated with Lac Z transduced HBC cells. Amplification of genomically incorporated Lac Z sequences in MDA-MB-231-BAG HBC cells enables us to specifically detect these cells in mouse organs and bones. We have also created a competitive template to use as an internal standard in the PCR reactions, allowing us to better quantitate levels of HBC metastasis. The results of this PCR detection method correlate well with cell culture detection from alternate long bones from the same mice, and are more sensitive than gross Lac Z staining with X-gal or routine histology. Comparable qualitative results were obtained with PCR and culture in a titration experiment in which mice were inoculated with increasing numbers of cells, but PCR is more quantifiable, less time consuming, and less expensive. This assay can be employed to study the molecular and cellular aspects of bone metastasis, and could easily be used in conjunction with RT-PCR-based analyses of gene products which may be involved with HBC metastasis.

Animals↗
PubMed1997

Production and characterization of a recombinant anti-MUC1 scFv reactive with human carcinomas.

Recombinant single-chain fragments (scFv) of the murine anti-MUC1 monoclonal antibody C595 have been produced using the original hybridoma cells as a source of variable heavy (V(H))- and variable light (V(L))-chain-encoding antibody genes. The use of the polymerase chain reaction (PCR), bacteriophage (phage) display technology and gene expression systems in E. coli has led to the production of soluble C595 scFv. The scFv has been purified from the bacterial supernatant by peptide epitope affinity chromatography, leading to the recovery of immunoreactive C595 scFv, which was similar in activity to the C595 parent antibody. Analysis by DNA sequencing, SDS-PAGE and Western blotting has demonstrated the integrity of the scFv, while ELISA, FACScan analysis, fluorescence quenching, quantitative immunoreactivity experiments and immunohistochemistry confirm that the activity of the scFv compares favourably with that of the parent antibody. The retention of binding activity to MUC1 antigen on human bladder and breast carcinoma tissue specimens illustrates the potential application of this novel product as an immunodiagnostic and immunotherapeutic reagent.

Amino Acid Sequence↗
PubMed1997

Effects of aerobic exercise training and yoga on the baroreflex in healthy elderly persons.

It is unclear whether the age-associated reduction in baroreflex sensitivity is modifiable by exercise training. The effects of aerobic exercise training and yoga, a non-aerobic control intervention, on the baroreflex of elderly persons was determined. Baroreflex sensitivity was quantified by the alpha-index, at high frequency (HF; 0.15-0.35 Hz, reflecting parasympathetic activity) and mid-frequency (MF; 0.05-0.15 Hz, reflecting sympathetic activity as well), derived from spectral and cross-spectral analysis of spontaneous fluctuations in heart rate and blood pressure. Twenty-six (10 women) sedentary, healthy, normotensive elderly (mean 68 years, range 62-81 years) subjects were studied. Fourteen (4 women) of the sedentary elderly subjects completed 6 weeks of aerobic training, while the other 12 (6 women) subjects completed 6 weeks of yoga. Heart rate decreased following yoga (69 +/- 8 vs. 61 +/- 7 min-1, P < 0.05) but not aerobic training (66 +/- 8 vs. 63 +/- 9 min-1, P = 0.29). VO2 max increased by 11% following yoga (P < 0.01) and by 24% following aerobic training (P < 0.01). No significant change in alpha MF (6.5 +/- 3.5 vs. 6.2 +/- 3.0 ms mmHg-1, P = 0.69) or alpha HF (8.5 +/- 4.7 vs. 8.9 +/- 3.5 ms mmHg-1, P = 0.65) occurred after aerobic training. Following yoga, alpha HF (8.0 +/- 3.6 vs. 11.5 +/- 5.2 ms mmHg-1, P < 0.01) but not alpha MF (6.5 +/- 3.0 vs. 7.6 +/- 2.8 ms mmHg-1, P = 0.29) increased. Short-duration aerobic training does not modify the alpha-index at alpha MF or alpha HF in healthy normotensive elderly subjects. alpha HF but not alpha MF increased following yoga, suggesting that these parameters are measuring distinct aspects of the baroreflex that are separately modifiable.

Aged↗