Search PubMed⌕ Search

Biomedical subjects

M Kaliner

Publications and source records attributed to M Kaliner.

At least 73 records · Page 4Linked to original sources
PubMed1984

Human pulmonary macrophage-derived mucus secretagogue.

Human pulmonary macrophages (PM) obtained from surgically removed human lung tissue released a factor after exposure to activated zymosan that caused cultured human airways to release increased amounts of radiolabeled mucous glycoproteins. The factor was released maximally after 4-8 h of zymosan exposure and caused a dose-related increase in glycoprotein release; it was termed macrophage-derived mucus secretagogue (MMS). MMS release was produced in a dose-dependent fashion by activated but not by nonactivated zymosan. The activation of zymosan was C3 dependent, and C3b-coated Sepharose was also an effective stimulant. The data suggested that cell surface activation of the PM was a sufficient stimulus to cause MMS release and that both C3-dependent activation as well as Fc receptor activation were effective. The synthesis of MMS was sensitive to cycloheximide, and no active MMS was detectable intracellularly. To determine if MMS might be one of the oxidative derivatives of arachidonic acid, PM were incubated with cyclooxygenase and lipoxygenase inhibitors before activation. These maneuvers did not influence MMS generation. MMS-rich supernatants were then extracted into organic solvents or exposed to lipophilic resin; in both cases, MMS remained in the aqueous phase. Thus, MMS is not a derivative of arachidonic acid. Sequential fractionation of MMS on ultramembrane and gel filtration followed by isoelectric focusing and gel filtration indicated that MMS is a small (approximately 2000 daltons), acidic (pI, 5.15) molecule. Therefore, surface activation of human PM results in the synthesis and release of a small acidic molecule that causes airway mucous glands to secrete increased quantities of mucous glycoproteins.

Animals↗
PubMed1984

Preparation of a human lung purified plasma membrane fraction: confirmation by enzyme markers, electron microscopy, and histamine H1 receptor binding.

A simple and rapid method of isolating plasma membranes from human peripheral lung tissue is described. The method involves homogenization of tissue in 0.25 M sucrose-buffered medium followed by differential and sucrose density gradient centrifugation. Enzymatic and morphological characterization of the plasma membrane fraction revealed minimal contamination by nonplasma membrane fragments. The isolated plasma membranes showed an 18-fold purification of 5'-nucleotidase activity compared to the original homogenate. Electron-microscopic studies of the plasma membrane fraction revealed the presence of small membrane vesicles having a trilaminar membrane structure. To further examine the purity of the plasma membrane preparation, the binding of the H1 receptor antagonist, 3H pyrilamine, to the plasma membrane-enriched fraction was compared to the binding to crude membrane preparations. Both the plasma membrane-enriched fraction and the crude membrane preparation had similar Kd's for the histamine antagonist, but the plasma membrane-enriched fraction had a threefold greater binding capacity, reflecting the relative enrichment of plasma membranes of the preparation. Thus, a method has been developed for the isolation of plasma membranes from human peripheral lung which should provide material for a variety of biochemical and pharmacological studies.

5'-Nucleotidase↗
PubMed1984

Prostaglandin-generating factor of anaphylaxis induces mucous glycoprotein release and the formation of lipoxygenase products of arachidonate from human airways.

The effects of prostaglandin-generating factor of anaphylaxis (PGF-A) upon the lipoxygenation of arachidonic acid and the promotion of mucous glycoprotein secretion by human airways were analyzed concurrently in order to determine the role that lipoxygenase products play in the secretion of mucus which accompanies immediate hypersensitivity reactions of airways. PGF-A enhanced both mucous glycoprotein release and the 5- and 15-lipoxygenation of arachidonic acid as well as the formation of leukotriene B4 (LTB4) with similar dose-response relationships. The capacity of PGF-A to stimulate mucous glycoprotein release was inhibited by ETYA but not by indomethacin, suggesting that PGF-A stimulated lipoxygenase products may be involved. Lipoxygenase products of arachidonic acid thus may serve as mediators of the enhancement of mucus secretion from human airways in response to PGF-A.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed1984

Demonstration that circulating human blood cells have no detectable alpha 1-adrenergic receptors by radioligand binding analysis.

The mechanisms underlying the autonomic nervous system abnormalities reported in allergic asthma have not been defined. In order to determine if these abnormalities reflect abnormal alpha-adrenergic receptor numbers or drug affinities, we attempted to identify alpha-receptors on circulating human blood cells. Platelets, red blood cells, polymorphonuclear leukocytes, and mononuclear cells were examined by use of radioligand binding techniques with the [3H]antagonists, dihydro-alpha-ergocryptine, prazosin hydrochloride, and yohimbine as ligands. The presence of alpha 2-receptors was confirmed on platelets, but no detectable alpha-receptors were identified on red blood cells or polymorphonuclear leukocytes. Preliminary observations suggested the presence of specific alpha-receptor binding to mononuclear cells; however, this binding was determined to reflect directly the presence of contaminating platelets. By use of a newly developed isolation technique to obtain platelet-depleted mononuclear cells, no alpha-adrenergic receptors could be identified on platelet-depleted mononuclear cells. Therefore, since no alpha 1-receptors could be identified on circulating human blood cells, these cells are not a suitable model for the study of the mechanisms underlying abnormal alpha-adrenergic responsiveness, and it may be necessary to reanalyze previous reports of alpha-adrenergic responsiveness on human blood cells with the use of platelet-depleted cell preparations.

Ergolines↗
PubMed1984

Induction of human cutaneous mast cell degranulation by opiates and endogenous opioid peptides: evidence for opiate and nonopiate receptor participation.

In order to examine the capacity of pharmacologically useful opiates to stimulate human mast cell secretion, subjects were skin tested with morphine, codeine, or meperidine hydrochloride. All three agents acted equipotently in eliciting positive immediate skin reactions from all subjects tested. Each agent demonstrated 10 mm of net whealing at 5 to 10 micrograms base (16.7 to 40.4 nmol) injected intradermally. The ability to elicit immediate skin test reactions with endogenous opioid peptides was examined with the use of dynorphin, [D-Ala, 2-D-Leu5] enkephalin, beta-endorphin, and morphiceptin . All four compounds induced wheal-and-flare reactions with the order of potency: dynorphin, greater than beta-endorphin, and greater than [D-Ala, 2-D-Leu5] enkephalin approximately equal to morphiceptin at dose ranges of 0.3 to 8.45 nmol. The inhibition of reactivity by hydroxyzine and the demonstration of mast cell degranulation by electron microscopy suggest that the immediate skin responses to opioid stimulation occur as a consequence of mast cell degranulation. Experiments with the opioid receptor antagonist, naloxone, suggest that both opioid and nonopioid receptors may be involved. These results imply that endogenous opioid peptides possibly may play a role in mast cell function and/or degradulation .

Codeine↗
PubMed1984

Heightened cholinergic responsiveness in IgE-producing rabbits.

Atopic humans express an array of autonomic nervous system abnormalities. An animal model of IgE-mediated allergy was herein employed to determine if the induction of an IgE antibody response would affect autonomic responsiveness. Rabbits were immunized so as to selectively produce IgE antibodies, and their pupillary miotic and mydriatic responses to cholinergic and alpha adrenergic stimuli, respectively, were determined. IgE-producing rabbits exhibited hyperresponsiveness to cholinergic but not alpha adrenergic stimulation, a pattern resembling that seen in nonasthmatic atopic humans. Repeated sublethal anaphylactic episodes induced in three IgE-producing rabbits did not affect the pattern of autonomic responsiveness when compared to controls challenged with saline, despite clear evidence of increased plasma histamine levels immediately after antigen challenges in the IgE group. This study indicates that elicitation of an IgE antibody response is associated with the concomitant development of cholinergic hyper-responsiveness.

Animals↗
PubMed1984

The effects of corticosteroids on mucous glycoprotein secretion from human airways in vitro.

In order to examine the mechanisms by which corticosteroids may benefit some patients with bronchorrhea, cultured human airways releasing [3H]glucosamine labeled mucous glycoproteins were exposed to corticosteroids, and mucus release was examined. Both dexamethasone and methylprednisolone produced dose-related suppression of the spontaneous release of radiolabeled mucous glycoproteins. The inhibitory effects of dexamethasone were maximal after 18 to 24 h and returned to control levels by 34 h. In order to study the effects of dexamethasone on stimulated mucus release, airways were exposed to dexamethasone and to the mucus secretagogues, histamine or 5-monohydroxyeicosatetraenoic acid. Both of these secretagogues stimulated radiolabeled mucous glycoprotein release from airways that had never been exposed to corticosteroids, as well as in a reduced fashion from corticosteroid-treated airways. The reduced mucus release caused by secretagogues from dexamethasone-treated airways appeared to reflect a lowered baseline secretion rate rather than a specific inhibition of either secretagogue.

Arachidonic Acids↗
PubMed1984

Production of mucous glycoproteins by nasal turbinates in vitro.

An in vitro system of human nasal turbinate tissue culture has been developed. Nasal turbinate tissue resected during surgery for nasal obstruction is dissected free of bone, placed on absorbable gelatin sponges, and cultured with CMRL-1066 medium containing antibiotics. Viability of explants may be demonstrated both physiologically and histologically through a period of 4 weeks. 3H-glucosamine added to the medium is biosynthetically incorporated into mucous glycoprotein (MGP). Gel filtration column chromatography on Sephacryl S-1000 in 6M urea in 0.005M phosphate buffer demonstrates human turbinate MGP to fractionate with 85% of the radiolabel filtered and 15% excluded by the column. The excluded MGP fractionates with globular proteins of greater than 20 X 10(6) daltons, while the fractions that enter the column filter with molecular sizes of 0.4 X 10(6) to 20 X 10(6) daltons. MGP synthesized by human lung airways has comparable sizing characteristics, suggesting a similarity in upper and lower airway mucus chemistry.

Culture Techniques↗
PubMed1984

Human respiratory mucous glycoproteins.

Biochemical characterization of human respiratory mucus has generally utilized expectorated specimens. In order to exclude extraneous contaminants in the analysis of airway glycoproteins, human airways were cultured and the mucous glycoprotein released into the supernatant analyzed. By incorporating 3H-labeled glucosamine or 14C-threonine into the media, the airways biosynthetically labeled the mucous glycoproteins (MGP), facilitating their analysis. The MGP chromatograph by gel filtration on Sepharose 2B in two fractions: one excluded from the column and one that enters the column. However, employing a gel filtration column with the ability to fractionate larger molecules, Sephacryl S-1000, it was found that MGP fractionate over a large range in molecular sizes and do not segregate into distinct fractions. The diffuse, broad peak of MGP fractionation on Sephacryl S-1000 is not affected by reduction and alkylation or by chromatography in 1 M NaCl. The fractionated MGP from Sepharose 2B were divided into larger and smaller molecular species, and their charge characteristics were determined by DEAE chromatography and preparative isoelectric focussing. MGP exhibit strong acidic charge characteristics that are uniform, as reflected in elution from DEAE and a single, sharp isoelectric focussing point. Enzymatic cleavage of the oligosaccharide side chains from MGP liberates more than 70% of the radiolabeled side chains. The side chains enzymatically cleaved from the larger and smaller molecular species of MGP are similar in size. Highly purified MGP were found to be 73% carbohydrate and 27% protein. Thus, human airways release a family of MGP that express marked heterogeneity in size but a uniform, strong acid charge and include side chains of similar size.

Carbohydrate Metabolism↗
PubMed1984

Inflammatory responses to mast cell granules.

Mast cell degranulation leads to classic allergic reactions appearing within minutes and abating in 30-60 min. It is now appreciated that this is but the first stage of a multisequenced reaction that includes late phase allergic reactions that are apparent within 4-8 h and persisting up to 24 h. These late phase reactions (LPR) are clinically experienced as burning, ill-defined erythema, and edema and are thought to participate in airway and nasal hyperreactivity. A rodent model has been developed that permits analyses of the pathogenesis of LPR: LPR may be elicited by isolated inflammatory factors released from mast cell granules, are complement independent, require an influx of neutrophils for full expression, and may be inhibited by combinations of H-1 and H-2 antihistamines as well as corticosteroids. The appreciation of the late phase of allergic reactions broadens our understanding of the pathologic features of immunologically induced inflammation as well as provides new approaches to the treatment of allergy and asthma.

Animals↗
PubMed1983

Detection of beta-adrenergic receptors on rabbit mononuclear cells isolated free of significant contamination by other cell types.

In order to study rabbit mononuclear cell surface receptors, it was necessary to develop a procedure to isolate mononuclear cell preparations that are free of significant contamination by other cell types, especially platelets. Centrifugation of dextran-sedimented, anti-coagulated whole blood through Hypaque (density 1.060) at 600 X g for 5 min at 22 degrees C eliminated greater than 93% of starting platelets. A second 5-min Hypaque centrifugation of Hypaque-Ficoll-isolated mononuclear cells (MNC) (approximately 80% lymphocytes) at 450 X g for 5 min at 22 degrees C reduced platelet contamination to less than one platelet per three MNC, and resulted in the overall removal of greater than 99.5% of starting platelets. These relatively pure MNC which were isolated in less than 2 hr were identified as having beta-adrenergic receptors by radioligand binding techniques using [125I]iodohydroxybenzylpindolol [( 125I]IHYP). Binding of [125I]IHYP to intact rabbit MNC was a saturable, stereospecific, and rapid process with a dissociation constant (KD) of 0.53 +/- 0.18 nM and a binding capacity of 3,461 +/- 235 sites/cell.

Animals↗
PubMed1983

Histamine H-1 binding site on human polymorphonuclear leukocytes.

Neutrophilic polymorphonuclear leukocytes (PMN) infiltrate the sites of allergic reactions and may respond to histamine, one of the major mediators of allergy. In order to characterize histamine interactions with PMN, the binding of [3H]pyrilamine was studied. Human PMNs bind [3H]pyrilamine in a specific, saturable, and reversible fashion and demonstrate specificity (H-1 antagonists greater than histamine greater than H-2 antagonists) for the competitive binding agents studied. Human PMNs have a homogeneous population of H-1 receptors of moderate affinity (Kd = 52 nM) in large number (265 X 10(3)/cell) which do not demonstrate cooperativity. Thus PMNs attracted to sites of allergic inflammation have H-1 binding sites which may respond to histamine stimulation.

Binding Sites↗
PubMed1983

Autonomic nervous system abnormalities in cystic fibrosis.

The basic defect in cystic fibrosis, the most common lethal genetic diseases of white Americans, is unknown, but the character of the generalized exocrinopathy suggests some disorder of the regulation and control of the process of glandular secretion. Definite abnormalities in all branches of the autonomic nervous system have been demonstrated in patients with cystic fibrosis, including increased sensitivity to alpha-adrenergically stimulated pupillary dilation; increased responsiveness to cholinergic stimulation of pupillary constriction, parotid saliva secretion, and eccrine sweat secretion; and decreased responsiveness to beta-adrenergic stimulation of the cardiovascular system as well as circulating lymphocytes and granulocytes. Since these abnormalities also occur in asymptomatic heterozygotes for cystic fibrosis (parents of patients), they are likely to be inherited characteristics and not secondarily acquired. This constellation of inherited autonomic abnormalities--alpha-adrenergic and cholinergic hyperresponsiveness and beta-adrenergic resistance--may contribute to the pathophysiology of cystic fibrosis and may also be an important clue to the nature of the basic defect.

Animals↗