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Biomedical subjects

S M MacDonald

Publications and source records attributed to S M MacDonald.

At least 37 records · Page 2Linked to original sources

Molecular identification of an IgE-dependent histamine-releasing factor.

An immunoglobulin E (IgE)-dependent histamine-releasing factor (HRF) produced by lymphocytes of atopic children and present in biological fluids of allergic patients has been identified and purified. Amino-terminal sequencing revealed extensive homology to a mouse protein, p21, and its human homolog, p23. Both recombinant proteins caused histamine release from the human basophils of a subpopulation of donors, and this release was dependent on IgE. Polyclonal antibodies recognized and removed the biological activity of recombinant and native HRF. HRF identifies a heterogeneity of IgE and is believed to play a prominent role in chronic allergic disease processes.

Amino Acid Sequence↗

Purification of immunoglobulin E (IgE) antibodies from sera with high IgE titers.

A three stage method for the ultrapurification of polyclonal IgE from human serum is reported using anion exchange chromatography followed by monoclonal antibody based positive and negative affinity chromatography. Following dialysis of 25-100 ml of serum (2.3-14 micrograms IgE/ml, n = 4) against 0.05 M Tris pH 8, each specimen was subjected to diethylaminoethyl (DEAE)-cellulose chromatography (serum/matrix = 1/4). IgE was eluted with 0.05 M Tris, 0.05 M NaCl pH 8, yielding an IgE recovery of 61-93%, with removal of approximately 90% of other serum proteins and an IgE purity ([IgE]/[Igs]) of 0.1-1.1%. After adjusting to 0.1 M NaCl and concentrating approximately 30-fold, the eluted IgE was further purified by affinity chromatography using a panel of IUIS/WHO-documented mouse monoclonal anti-human immunoglobulin antibodies (alpha hIg-MAbs). First, the IgE-enriched DEAE-cellulose chromatography fraction was incubated in a batch mode with two alpha hIgE-Fc MAbs (HP6029, HP6061) coupled to CNBr-Sepharose, CL-4B. IgE was eluted with 0.05 M glycine pH 2.8 and immediately neutralized. The IgE recovery was 32-52% and IgE purity was 72-97%. Silver-stained SDS-PAGE and noncompetitive solid-phase two-site immunoenzymetric assays for total human IgA, IgE, IgG and IgM indicated that IgA, IgG and IgM were the only contaminants. Next, the IgE was concentrated 10-30-fold in the presence of 0.1% HSA. One IgE specimen was ultrapurified in a batch mode by negative selection chromatography using three pairs of alpha hIg-MAbs (alpha hIgA: HP6111 + HP6123; alpha hIgG: HP6017 + HP6046; alpha hIgM: HP6081 + HP6083) coupled to CNBr-Sepharose, CL-4B. IgE purity increased from 91% to > 99.9% with approximately 70% recovery of IgE for this step. The ultrapurified IgE antibody was shown to be functionally reactive for allergen and Fc epsilon RI receptors on human basophils. We conclude that alpha hIg-MAbs are powerful tools to facilitate the affinity purification of functionally active human IgE from serum; however, when the analyte is present in low concentration, a carrier protein needs to be added to minimize non-specific loss of the material during this process.

Antibodies, Anti-Idiotypic↗

Hypercalcemia of malignancy in the palliative care patient: a treatment strategy.

Hypercalcemia of malignancy is most commonly due to the effects of parathyroid hormone-related peptide, which acts as a humoral factor to cause generalized osteoclast-mediated bone resorption and reabsorption of calcium by the kidney tubule, and may also act as a local resorptive factor adjacent to bone metastases. Local resorptive mechanisms are less common causes of malignant hypercalcemia than previously believed. Treatment begins with intravenous fluid rehydration, followed by a furosemide diuresis and the bisphosphonate pamidronate, 60-90 mg, intravenously. Gallium nitrate is an efficacious but inconvenient alternative to pamidronate. Calcitonin combined with pamidronate is a reasonable initial therapy for severe hypercalcemia to hasten normalization of the serum calcium. Steroids should be reserved for hypercalcemia due to tumor production of 1,25 dihydroxyvitamin D, or for steroid-responsive malignancies. Oral or parenteral bisphosphonates can be used to maintain normocalcemia. In addition to improving the morbidity of acute hypercalcemia, bisphosphonate therapy has been shown to reduce bone pain and pathological fractures in patients with bone metastases, and calcitonin also has a potent analgesic effect in these patients. Treatment for hypercalcemia should therefore be considered in the majority of patients in the palliative care setting.

Aged↗

The assessment of constipation in terminal cancer patients admitted to a palliative care unit: a retrospective review.

Constipation is a frequent and distressing complication in patients with advanced cancer. However, very few studies have reviewed the assessment and management of these patients. The purpose of this study was to review the documentation and assessment and diagnosis of constipation in patients admitted to a Palliative Care Unit, and the correlation between those findings and radiological evidence of stool in the colon. The records of 122 consecutive patients admitted to the Palliative Care Unit, Edmonton General Hospital were reviewed in order to assess the physician's and the nurse's record of symptoms, physical findings, and diagnosis and treatment of constipation. All patients also underwent a flat abdominal radiograph that scored for the presence of stool in the colon (0 = no stool; and 12 = stool occupying all the lumen of the four quadrants of the colon). The radiograph was scored blindly by two different physicians. Of 103 evaluable patients, a rectal exam was reported only in 42. Correlation between the assessment by the two physicians' radiograph score was high (0.78, P nd nurses' diagnosis of constipation, the presence of laxative treatment, the number of days since the last bowel movement, and the source of the admission (hospital vs home) were not associated with higher radiological scores for constipation. Assessment is insufficient in this population at high risk for severe constipation. Radiological examination may be necessary for adequate diagnosis in some patients. More research is needed in this area.

Aged↗

Proximal muscle weakness in a patient with hepatocellular carcinoma.

Asthenia and generalized weakness are common in cancer patients. There are multiple causes for these symptoms. We describe a case of rapid onset of proximal muscle weakness in a patient with hepatocellular carcinoma. The differential diagnosis of muscle weakness in the palliative care patient is reviewed. The discussion centers on steroid myopathy and its treatment.

Asthenia↗

Screening with total cholesterol: determining sensitivity and specificity of the National Cholesterol Education Program's guidelines from a population survey.

The objective of the study was to determine the sensitivity and specificity of the National Cholesterol Education Program (NCEP) guidelines in identifying at-risk individuals for hypercholesterolemia, using population-based data from the Manitoba Heart Health Survey (MHHS). The MHHS surveyed a representative sample of adult residents of the province of Manitoba, Canada, aged 18-74 (n = 2792), 2212 of whom underwent complete lipoprotein analyses: total cholesterol (TC), high-density (HDL) and low-density (LDL) lipoprotein cholesterol, and triglycerides (TG) after overnight fasting. Following NCEP criteria for risk categories, 618 individuals could be considered as "at risk" based on their HDL and/or LDL levels. However, if the NCEP algorithms were followed, i.e. initial screen for TC only, and determination of HDL and LDL for selected individuals with borderline and high TC, only 438 of the total "at risk" individuals would have been identified (sensitivity = 71%). 1203 of 1594 individuals with acceptable HDL and LDL levels were not considered "at risk" on screening (specificity = 76%). If "at risk" status was determined solely on LDL criteria, the NCEP guidelines result in a higher sensitivity (94%) and comparable specificity (76%). However, when only HDL criteria were used, the sensitivity and specificity declined to 38% and 63% respectively. Compared to a simulated screening study based on the Lipid Research Clinic data reported by Bush and Riedel (1991) the estimates of sensitivity from the MHHS were high. Current NCEP guidelines appear to balance optimally the sensitivity and specificity for selecting individuals based on TC levels for further complete lipoprotein analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

The heterogeneity of human IgE exemplified by the passive transfer of D2O sensitivity.

Basophil responsiveness to histamine-releasing factors (HRF) is limited to cells from atopic donors; this response can be transferred passively to non-reactive basophils by IgE molecules from the sera of donors who are intrinsically responsive to HRF. Deuterium oxide (D2O) also causes mediator release from the basophils of atopic asthmatic subjects. To assess whether basophil responsiveness is IgE dependent, and, if so, whether this release revealed IgE heterogeneity, we tested the ability of sera from HRF responders (IgE+) and non-responders (IgE-) to sensitize basophils to D2O. Both purified IgE+ and unpurified sera from an HRF responder were passively used to sensitize basophils whose IgE had been removed by lactic acid treatment. As a control, an IgE- myeloma-containing serum was used for passive sensitization. In five experiments, histamine release in the presence of 44% D2O was 9 +/- 2% and 46 +/- 4% using control IgE- and IgE+ sensitized cells, respectively. The non-responder serum, even at higher IgE levels, did not sensitize the cells for D2O release. If the IgE receptors on lactic-acid treated cells were first exposed to serum from an IgE- donor, sensitization to D2O by IgE+ was blocked. The percentage histamine release to D2O was directly related to both the amount of IgE+ used for passive sensitization and the concentration of D2O used for release. These experiments further support the concept of IgE heterogeneity and suggest that the occupancy of IgE receptors on the basophil surface 'activate' the cell to make it more responsive to various stimuli.

Antigens, Differentiation, B-Lymphocyte↗

IgE-dependent histamine-releasing factors. A brief review.

A cytokine, termed histamine-releasing factor (HRF) and produced by many cell types, has become the focus of research by many investigators due to its potential importance as a stimulus in chronic inflammation. We are producing and characterizing an HRF which causes IgE-mediated histamine release from human basophils. Following extensive purification procedures, the molecule will be sequenced and synthesized. A functional heterogeneity of IgE molecules was revealed by these studies. We are currently producing IgE antibody in vitro and testing the hypothesis that differential glycosylation is the basis for the heterogeneity. Knowledge of the structures and interactions of these molecules should advance our understanding of allergic and more chronic diseases.

Basophils↗

Histamine-releasing factors and heterogeneity of IgE.

The duration and severity of the allergic response are variable. Even though antigens are rapidly cleared from the individual, an acute allergic response is frequently followed by a recrudescence of symptoms hours or even days after the initial exposure. Experimentally, the cellular infiltrates and mediators released during this late response resemble those associated with chronic inflammatory disease. Although basophils are present in this late reaction, the stimuli for their activation remain unknown. A heterogeneous group of unique cytokines called histamine-releasing factors (HRF), discovered over a decade ago, may well play a role in stimulating basophils during this late-phase reaction. These factors have been reported from a variety of cell sources including alveolar macrophages, platelets, vascular endothelial cells, B and T lymphocytes, mononuclear cell cultures, the U937 monocyte/macrophage-like cell line and the RPMI 8866 B cell line. These ubiquitous factors cause non-cytotoxic, calcium-dependent mediator release from human basophils in vitro and are also present and active in vivo. Purification attempts have revealed that HRF exists in at least three forms, based on molecular weight. In our hands, the mechanism of mediator release by one of the forms of HRF is IgE dependent. Since only about 50% of allergic donors' basophils respond to HRF, a heretofore unappreciated heterogeneity of IgE was revealed. The presence of HRF has been shown to correlate with severity of allergic disease in children with food allergies, with symptoms in the late-phase response in adults and with severity of the allergic response to an inhaled antigen. Thus, the study of HRF has evolved over the last decade and may lead to better understanding of the complex allergic response.

Cytokines↗

Recombinant human IL-1 alpha and -1 beta potentiate IgE-mediated histamine release from human basophils.

In this study, we have explored the relationship between interleukins and human basophil activation. Previous studies by ourselves and others have found that recombinant human (rh) IL-3 causes histamine release. The ability to release histamine has also been claimed for IL-1 but we cannot confirm this. In experiments with the basophils of 29 donors (excluding one D2O responder), histamine release with 100 ng/ml rhIL-1 alpha was 1.3 +/- 1% (SEM), whereas with rhIL-1 beta, it was 0.8 +/- 1%. Both IL-1 alpha and -1 beta were also used at concentrations of 0.01 to 1000 ng/ml without causing release. Neither increasing the Ca2+ concentration nor adding D2O or cytochalasin B caused IL-1 alpha and -1 beta to become secretagogues. rhIL-1, however, did augment IgE-dependent histamine release. The enhancement was similar with both rhIL-1 alpha and -1 beta, i.e. they were dose-dependent between 0.1 and 3 ng/ml and reached a plateau from 3 to 100 ng/ml. At submaximal histamine release (less than 10%), there was enhancement of three IgE-dependent secretagogues: 125% with goat anti-human IgE (n = 7), 215% with Ag E (n = 10), and 260% with a histamine releasing factor (n = 7). Non-IgE-dependent stimuli (formyl-methionine-leucine-phenylalanine and the ionophore A23187, n = 10) were enhanced less than 5%. rhIL-1-enhancement persisted after cell washing (n = 10). rhIL-1 was active in preparations of 50 to 75% pure basophils in which mononuclear cells were reduced by greater than 95% (n = 4), and mAbH34 to IL-1 beta blocked the enhancement caused by that molecule. We postulate that basophils have an IL-1 receptor which, when occupied, upregulates the response to IgE-related signals. Thus, this work characterizes a second interaction between interleukins and the cells central to the allergic response.

Adjuvants, Immunologic↗

Recombinant IL-3 induces histamine release from human basophils.

Human rIL-3 induces histamine release from some human basophils, with cells from atopics responding to a greater extent than non-atopic donors. The dose response curves were highly variable. IL-3 was active on purified basophils and the release process was slower and required more calcium than anti-IgE. Removal of surface IgE from basophils rendered them unresponsive to IL-3. The response could be restored by passive sensitization of basophils with IgE+, IgE known to bind histamine-releasing factors, and not IgE-, IgE unreactive with histamine-releasing factors. Thus, IL-3 uncovers IgE heterogeneity. IL-3 does not, however, directly interact with IgE+. Rather, passive sensitization with IgE+ or stimulation of basophils with low concentrations of several secretagogues renders the cells sensitive to IL-3. IL-3 may well play a pro-inflammatory role by potentiating the effects of IgE+ or various secretagogues.

Antibodies, Anti-Idiotypic↗

Cells and secretagogues involved in the human late-phase response.

Those scientists interested in allergic inflammatory processes have recently been focusing on the late-phase response, since it appears most similar to the chronic disease states observed in allergic patients. In this review we will focus on the pattern of mediator release and cellular traffic observed in two in vivo human models of the late-phase reaction, one involving the upper airways and the other the skin. We have observed in these models, as had been observed earlier in blood, that the late-phase reaction is associated with a second increase in the level of mediators. We also describe our studies of the secretagogues responsible for this late-phase mediator release and, in so doing, introduce the subjects of histamine-releasing factors and IgE heterogeneity.

Adrenal Cortex Hormones↗

Studies of IgE-dependent histamine releasing factors: heterogeneity of IgE.

Nasal lavage fluids from unstimulated individuals contain a histamine-releasing factor (HRF) similar to those which we have previously described from macrophages, platelets, and from blister fluids obtained during the late cutaneous reaction. The nasal HRF was partially purified by ion-exchange chromatography and gel filtration. Although some m.w. heterogeneity was observed, the majority of the HRF eluted at an apparent m.w. range of 15,000 to 30,000. This partially purified HRF induced histamine release from basophils of certain individuals. Histamine release occurred via a mechanism which is IgE-dependent in that: basophils desensitized by exposure to anti-IgE in the absence of calcium no longer respond to HRF, and desensitization with HRF reduces responsiveness to anti-IgE; and removal of IgE from the basophil surface by using lactic acid renders cells unresponsive to HRF. We have further defined this IgE dependence and have shown that the reason that only selected basophil donors respond to HRF is due to a previously unrecognized, functional heterogeneity of IgE. Thus, passive sensitization using sera from responders restored the responsiveness of acid-stripped basophils and conferred responsiveness to basophils of a nonresponder with naturally unoccupied IgE receptors. Sera from nonresponders failed to do this even though similar numbers of IgE molecules were put onto the basophil surface in each case. This property of responder sera was due to IgE because both heating sera at 56 degrees C for 2 hr and passage of sera over anti-IgE-Sepharose (which removes greater than 90% of the IgE) markedly reduced the ability of sera to induce responsiveness, and because an excess of either purified IgE myeloma or purified penicillin-specific IgE antibody from a nonresponder competitively inhibited the ability of IgE from responder sera to induce responsiveness to HRF. We conclude that nasal lavage fluids contain an HRF which induces basophil histamine release in a specific, IgE-dependent fashion but only from individuals with the appropriate type of IgE. Because we have shown that basophils are recruited into the nose during the late-phase reaction, we suggest that nasal HRF may induce these cells to release histamine and other mediators which could contribute to the symptomatology of the late-phase reaction.

Basophils↗

Levels of cadmium, mercury and lead in otter and mink faeces from the United Kingdom.

Over 500 faeces of otter (Lutra lutra) from Britain, with a small sample from northeast Greece, were analyzed for Cd, Hg and Pb. A small sample of mink (Mustela vison) faeces was also analyzed. Mean concentrations of metals (mg kg-1 dry weight) in otter faeces ranged from 1.53 to 3.97 Cd, 0.25 to 0.74 Hg and 12.4 to 20.9 Pb. Mink faeces generally contained less Cd and Hg, but more Pb. Metal concentrations in faeces may reflect both natural and anthropogenic sources. Estimates of daily Cd and Pb intake were made from faecal concentrations and these were in good agreement with concentrations in major fish prey. The estimates of daily Cd and Pb intake are high, but nevertheless most samples came from thriving otter populations. The value of faecal analysis in assessing metal burdens during surveys of otters is discussed.

Animals↗

A monoclonal antibody recognizing the p150/95 leucocyte differentiation antigen.

A monoclonal antibody KB90 recognizing a molecule containing two polypeptide subunits of 140 and 98 kd is described. Biochemical studies following high pH dissociation of the molecule showed KB90 to bind to an epitope on the 140 kd subunit. The molecule recognized by KB90 appears to correspond to the p150/95 molecule in the family of leucocyte differentiation antigens of which the LFA-1 and Mac-1 molecules are members. Analysis of the immunocytochemical reactivity of KB90 shows that it stains monocytes, tissue macrophages, granulocytes and a small population of lymphocytes. These reactions are clearly distinct from those of the LFA-1 and Mac-1 molecules.

Animals↗