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H M Brown

Publications and source records attributed to H M Brown.

At least 37 records · Page 2Linked to original sources
PubMed1990

Dimethyl sulfoxide elevates intracellular Ca2+ and mimics effects of increased light intensity in a photoreceptor.

A 1% (v/v) solution of dimethyl sulfoxide (DMSO) added to the saline bath of isolated Balanus eburneus photoreceptors increased receptor potential amplitude by 40-50% and shortened time to peak amplitude and latency by 20-25%. The light-sensitive membrane current of voltage-clamped cells was increased systematically as DMSO concentration was increased from 1% to 10%. The null potential of the light sensitive current was unaffected by DMSO with short pulses of light, indicating that DMSO has no direct effect on ion selectivity of the light-sensitive channel. Absorbance changes of cell injected with the calcium indicator arsenazo III show that DMSO elevates intracellular Ca2+ (Cai). Current-voltage relations in darkness reveal that DMSO induces a small sustained inward current (approximately 5 nA) which has a null potential similar to the light-induced current. DMSO may activate the light-sensitive conductance via the increase in Cai. However, the altered kinetics and increased amplitude of the receptor current are opposite to the desensitizing effects normally observed with increased Cai.

Animals↗
PubMed1989

Effect of low-density lipoprotein on the expression of high affinity Fc gamma receptors.

A substrain of the human monocyte-like cell line U937, which is a cholesterol auxotroph, was used to study the effect of cellular cholesterol depletion on the expression of the type I Fc receptor for IgG (Fc gamma RI). Measurement of Fc gamma RI expression was performed by immunofluorescence and flow cytometry using the monoclonal antibody (mAb) 32.2, which is specific for an epitope on Fc gamma RI, and monomeric IgG2a, which binds to the ligand binding site of Fc gamma RI. Incubation of these cells for 24 h in growth medium containing delipidated fetal calf serum depletes cellular cholesterol without affecting growth or viability. While incubation of U937 cells with human interferon-gamma (IFN-gamma) increased Fc gamma RI expression, cholesterol depletion after cell growth in media containing delipidated serum and IFN-gamma resulted in reduced binding of both mAb 32.2 and IgG2a. A significant decrease in the number of cell surface binding sites, as measured by mean fluorescence intensity, was observed after cholesterol depletion. Supplementation of the delipidated serum medium with pure cholesterol in an ethanol/bovine serum albumin mixture, which replenished cellular cholesterol and supported growth, failed to restore antibody binding significantly. In contrast, low-density lipoprotein (LDL) which also delivered cholesterol to the cells restored binding both in terms of the number of the reactive cells and cell surface receptor density. High-density lipoprotein (HDL3), which does not deliver cholesterol to the cells, showed results similar to those obtained with pure cholesterol. This indicates that either LDL cholesterol is better utilized for membrane synthesis than pure cholesterol or that LDL provides another component, in addition to cholesterol, which is required for expression of Fc gamma RI, but not for growth. These studies indicate a role for LDL in regulating the expression of Fc gamma RI on the cell surface.

Antibodies, Monoclonal↗
PubMed1988

Intracellular calcium changes in Balanus photoreceptor. A study with calcium ion-selective electrodes and Arsenazo III.

Intracellular Ca2+ concentration (Cai) in the dark and during light stimulation, was measured in Balanus photoreceptors with Ca2+ ion-selective electrodes (Ca-ISE) and Arsenazo III absorbance changes (AIII). The average basal Cai of 17 photoreceptors in darkness was 300 +/- 160 nM determined with liquid ion-exchanger (t-HDOPP) Ca-ISE. Ca-ISE measurements indicated that light increased Cai by 700 nM (average), whereas AIII indicated an average change of 450 nM. The time course of AIII absorbance changes matched the time course of changes in the receptor potential more closely than did the Ca-ISE. Changes in Cai were graded with light intensity but the change in Cai was much greater for a decade change in intensity at high light intensity than at low intensity. The peak light induced conductance change of voltage clamped cells had a relationship to light intensity similar to that of the change in Cai. The peak Cai level measured with Ca-ISE was in good agreement with the free Ca2+ concentration of injected buffer solutions. Control Cai levels were usually restored within 5 min following injection of Ca2+ buffers. Injection of Ca2+ buffers with free Ca2+ of 0.6 microM produced a membrane depolarization. Larger increases in Cai (greater than microM) produced by injection of CaCl2 or release of Ca2+ from injected buffers by acidifying the cell, produced a pronounced membrane hyperpolarization. Increasing Cai with all of these techniques reduced the amplitude of the receptor potential. The time course of the receptor potential recovery was usually similar to that of Cai recovery.

Animals↗
PubMed1987

A requirement for cholesterol for phorbol ester-induced adhesion of a human monocyte-like cell line.

The human monocyte/macrophage-like cell line U937, which is a cholesterol auxotroph, is nonadherent. However, it becomes adherent after treatment with phorbol 12-myristate 13-acetate (phorbol ester). We investigated the effects of cellular cholesterol depletion and repletion on the effectiveness of phorbol ester to induce adhesion to substratum. Almost 70% of cellular cholesterol is depleted by incubation of the cells for 24 hrs in the growth medium in which delipidated fetal calf serum is substituted for fetal calf serum without affecting viability or the rate of growth. The use of delipidated fetal calf serum inhibited phorbol ester-induced adhesion by 40%. If the cells were preincubated in the medium containing delipidated fetal calf serum 6 hrs prior to addition of phorbol ester, adhesion was inhibited by 90%. Addition of cholesterol to the medium containing delipidated fetal calf serum, which replenishes cellular cholesterol, restored the ability of phorbol ester to induce adhesion to levels seen in cells cultured in the medium containing fetal calf serum. Epicholesterol was not as effective as cholesterol in supporting adhesion. Cholesterol depletion did not inhibit phorbol ester stimulation of superoxide anion production. These observations indicate a function for cholesterol in phorbol ester-induced adhesion that is independent of phorbol ester-induced superoxide anion production. It is proposed that cholesterol is required for synthesis and/or proper orientation and distribution, in the plasma membrane, of macromolecule(s) that mediate phorbol ester-induced adhesion.

Cell Adhesion↗
PubMed1987

Skin testing.

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Allergens↗
PubMed1987

Taking a history.

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Employment↗
PubMed1986

The cellular basis for the induction of antigen-specific T8 suppressor cells.

The cellular basis for the generation of antigen-specific T8 suppressor cells with high doses of antigen has been studied. We separated the T4 subset of human T cells into T4+2H4+ and T4+2H4- subpopulations with a recently developed monoclonal anti-2H4 antibody. T8 cells could be consistently activated to suppress a primary anti-2,4-dinitrophenyl (DNP) antibody response in vitro with unfractionated T4 cells or with the T4+2H4+ subset but not the T4+2H4- subset. In contrast, the T4+2H4- subset functioned as the helper inducer for the anti-DNP antibody response. With keyhole limpet hemocyanin (KLH)-stimulated T4+2H4+ cells we could efficiently induce antigen-specific suppressor activity of fresh T8 cells. In contrast, the T4+2H4+ subset could not effect suppression in the absence of T8 cells. Our findings indicate that the T4+2H4+ subset of human T cells is the suppressor inducer of specific T8 cells in an antigen-specific DNP-KLH system.

Antibodies, Monoclonal↗
PubMed1986

Study of IgG sub-class antibodies in patients with milk intolerance.

An ELISA was applied to measure IgG sub-class antibodies to cow's milk beta-lactoglobulin (BLG), alpha-lactalbumin (ALA) and alpha-casein (AC) and to hen's egg ovalbumin (OA) in the sera of nineteen adult patients with milk intolerance causing either asthma, eczema or both. Results were compared with those of forty blood donors and twenty adult patients with either asthma or eczema due to inhalant allergy. Apart from one blood donor, high titres of IgG sub-class antibodies to all three milk proteins were found only in the milk intolerance group. The most frequently detected antibody was AC-specific IgG4; being high (i.e. greater than 9.98 micrograms/ml) in eight milk intolerance cases: six with eczema, one with asthma and one with both. A variable proportion of these eight patients also had high levels of IgG1, IgG2 and IgG3 antibodies to AC and IgG1, IgG2, IgG3 and IgG4 antibodies to BLG and ALA. In contrast, IgG antibody to the egg protein, OA, was remarkably restricted to IgG4 and was present in high titres in 68.4% of milk intolerant patients, 60% of inhalant allergy patients and 30% of blood donors. However, the greater incidence of high titres of IgG4 antibody to OA, compared to AC, was due to the superior coating efficiency of OA resulting in a more sensitive assay. We conclude that some adult cases of milk intolerance, particularly those with eczema, can be diagnosed by detecting raised serum levels of IgG sub-class antibodies to milk proteins.

Adolescent↗
PubMed1986

Intracellular free Mg2+ in Balanus photoreceptor measured with eriochrome blue.

Intracellular free Mg2+ (Mgf) in darkness and during illumination was measured in the Balanus photoreceptor using the Mg2+ sensitive dye Eriochrome Blue SE (EB). The EB was pressure injected in the cell and Mgf determined from measurements of absorption changes at 570 nm and 600 nm. Mgf is 1.7 mM in the dark-adapted photoreceptor. There was no indication of a change in Mgf when the cell was illuminated with bright light, indicating that Mgf is not elevated during the phototransduction. Changes less than 5 X 10(-5) M could have occurred and remained undetected. The spectrophotometric properties of the indicator were studied at 10(-4) and 10(-3) M, at ionic conditions relevant to the intracellular measurements. Both the differential molar absorption coefficient for the EB-Mg complex at 600 nm (delta epsilon 600EBMg) and the apparent dissociation constant (KD) for the complex formation were different at 10(-4) and 10(-3) M EB.

Animals↗