PubMed Health⌕ Search

Biomedical subjects

E J Goetzl

Publications and source records attributed to E J Goetzl.

At least 19 recordsLinked to original sources
PubMed1992

An acidic fibroblast growth factor protein generated by alternate splicing acts like an antagonist.

Polymerase chain reaction amplification of cDNA for acidic fibroblast growth factor in several lines of cultured human cells revealed two forms of mRNA. The novel smaller mRNA lacks the entire second coding exon of the acidic fibroblast growth factor gene, whereas the previously identified mRNA consists of three coding exons. The truncated variant of acidic fibroblast growth factor (aFGF') is only 60 amino acids long with an apparent molecular mass of 6.7 kD on sodium dodecyl sulfate gels in contrast to 18 kD for the full-length acidic fibroblast growth factor. aFGF' elicits only minimal fibroblast proliferation and antagonizes the effects of acidic fibroblast growth factor when added exogenously to or when coexpressed with aFGF in BALB/c/3T3 fibroblasts. Thus, the truncated variant of acidic fibroblast growth factor may provide fibroblasts with a unique mechanism for endogenous regulation of their responses to acidic fibroblast growth factor.

Amino Acid Sequence↗
PubMed1992

Ligand-induced formation of the leukotriene B4 receptor-G protein complex of human polymorphonuclear leukocytes.

The components of the polymorphonuclear leukocyte (PMNL) receptor for leukotriene B4 (LTB4) were examined by Sephacryl S-300 exclusion chromatography of PMNL membrane proteins, which were solubilized before and after the binding of [3H] LTB4. When the PMNL membranes were solubilized in 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) and filtered on Sephacryl S-300 prior to addition of [3H] LTB4, the binding activity was associated with a 65 kD protein. In contrast, the radioactivity of [3H] LTB4 bound to PMNL membranes prior to solubilization was recovered predominantly with a 140 kD protein. When PMNL membranes had been pretreated with pertussis toxin, but not cholera toxin, before the addition of LTB4 and subsequent solubilization, radioactivity was recovered predominantly with the 65 kD protein. The addition of guanylylimidodiphosphate (GMP-PNP), a nonhydrolyzable derivative of guanosine triphosphate (GTP), to PMNL membrane receptors bearing [3H] LTB4 either prior to or after CHAPS solubilization reduced the yield of the 140 kD presumed LTB4 receptor protein-G protein complex. That the maximum specific binding of [35S] guanosine-5'-0-3-thiotriphosphate (GTP-gammaS) to LTB4-binding proteins in the Sephacryl S-300 effluent corresponded to the 140 kD protein supported the presence of a G protein in the LTB4 receptor complex.

Cell Membrane↗
PubMed1992

Neuropeptides in pulmonary edema fluid of adult respiratory distress syndrome.

A role for peptidergic nerves in the adult respiratory distress syndrome (ARDS) was examined by radioimmunochemically quantifying neuropeptides in pulmonary edema (PE) fluids from seven patients with ARDS and six patients with PE from congestive heart failure (CHF). The PE fluid mean concentrations of substance P (SP) and gastrin-releasing peptide (GRP) were significantly higher in ARDS (0.59 +/- 0.29 SD and 0.10 +/- 0.03 nM, respectively, P < 0.001 for both) than in CHF (0.19 +/- 0.08 and 0.04 +/- 0.01), whereas no difference was detected between the mean levels of vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) in the two forms of PE. Mean alveolar fluid concentration of SP was 8.7 nM (range 2.1-20.5 nM, N = 4) in sheep with acute lung injury from intravenous Pseudomonas aeruginosa, but was undetectable in sheep with balloon-induced high left atrial pressure simulating CHF (N = 2) or control sheep (N = 2). Pulmonary lymphatic clearance of SP, which reflected the rate of generation of SP in the lungs, attained a maximum of 25-95 pmol/h in sheep given P. aeruginosa intravenously, but was detected in only one of four control sheep at a lower level. Some pulmonary neuropeptides thus are released locally by acute lung injury and may contribute to endothelial and/or epithelial abnormalities underlying the altered capillary-alveolar permeability in ARDS.

Adult↗
PubMed1992

Mediators of communication and adaptation in the neuroendocrine and immune systems.

Bidirectional interactions between the immune and neuroendocrine systems influence specifically physiological activities as diverse as tissue localization of lymphocytes, antibody responses, hypothalamic-pituitary hormone secretion, and neural signal transmission. Our understanding of intersystem communications has been increased by the delineation of the innervation of immune organs, effects of neuromediators on immune cells, and neuroendocrine responses to individual immune cytokines. Two patterns of responses to neuroimmune mediators have been defined to distinguish between direct alterations in cellular function and the more complex states of differentiation and adaptation that condition the threshold and nature of reactions to subsequent stimuli. Recent identification of the molecular mechanisms of action of neuroimmune mediators is exemplified by elucidation of the requirement for prolactin in T lymphocyte proliferation and of the effects of vasoactive intestinal peptide on B lymphocyte expression of adherence proteins. Further knowledge of the advantages of multisystem integration of functions in host defense may reveal other novel mechanisms of cellular communication and biological adaptation.

Cell Communication↗
PubMed1992

Eicosanoids, but not tachykinins, excite C-fiber endings in rat sciatic nerve-end neuromas.

Normal nociceptors are sensitized by hyperalgesic mediators such as eicosanoids and tachykinins. The possibility that these mediators contribute to hyperalgesic pain associated with neural injury was investigated by examining their effects on the excitability of injured afferent nerve endings. In amounts that sensitize normal nociceptors and are hyperalgesic in normal skin, the eicosanoids prostaglandin I2 (PGI2), and 8(R),15(S)-dihydroxyicosatetraenoic acid (8(R),15(S)-diHETE) both excited some C-fibers in chronic neuromas of rat sciatic nerve. In contrast, the selective tachykinin-receptor agonists septide and senktide did not excite C-fibers. None of the mediators affected A-fibers. We conclude that PGI2 and 8(R),15(S)-diHETE may contribute to post-injury pain and hyperalgesia by an action on injured afferent endings.

Animals↗
PubMed1992

Mouse monoclonal antibody to a latent epitope of leucocyte receptors for leukotriene B4.

Human blood polymorphonuclear (PMN) leucocytes and human leucocytes of the HL-60 line, which were induced to differentiate by 1,25-dihydroxyvitamin D3, express stereospecific receptors for the potent chemotactic mediator, leukotriene B4 (LTB4), that is derived by 5-lipoxygenation from arachidonic acid. Monoclonal antibodies to LTB4 receptors (LTB4-R) were generated by immunizing BALB/c mice with partially purified PMN leucocyte membrane proteins, and fusing their splenocytes with P3X63Ag8 mouse myeloma cells. Hybridoma supernatants were screened initially by binding to PMN leucocyte LTB4-R protein, which had been affinity cross-linked with aminopropylamide (APA)-LTB4 and immobilized in plastic wells through attachment of the linked APA-LTB4 to adherent Fab of monoclonal anti-LTB4. Of the three clones producing antibodies which bound to LTB4-R, 0.5 mg/ml of one IgG3k antibody, termed E2, precipitated over 90% of the [3H]LTB4-binding activity of solubilized PMN leucocyte membrane proteins. E2 also bound to a radiolabelled protein of 70,000-80,000 MW from 125I-labelled PMN leucocyte membranes [35S]-labelled HL-60 cell membranes, and PMN leucocyte membranes affinity-labelled with [3H]APA-LTB4, that was identical in size to the LTB4-R precipitated by the rabbit IgG anti-idiotypic antibodies. E2 did not bind to intact PMN leucocytes or modify the binding of [3H]LTB4 by PMN leucocytes. The binding of E2 to LTB4-R in purified membranes of PMN leucocytes was less than one-fourth of that observed for the anti-idiotypic antibodies, but increased substantially after solubilization of the LTB4-R. The E2 monoclonal antibody thus recognizes a partially latent substituent of LTB4-R, which does not contribute to combining site function.

Animals↗
PubMed1991

Cloning and expression of the human vasoactive intestinal peptide receptor.

Vasoactive intestinal peptide (VIP) is a neuroendocrine mediator found in the central and peripheral nervous system. Distinct subsets of neural, respiratory, gastrointestinal, and immune cells bear specific high-affinity receptors for VIP, which are associated with a guanine nucleotide-binding (G) protein capable of activating adenylate cyclase. A cDNA clone (GPRN1) encoding the human VIP receptor was identified in libraries prepared from the Nalm 6 line of leukemic pre-B lymphoblasts and the HT-29 line of colon carcinoma cells. The deduced 362-amino acid polypeptide sequence encoded by GPRN1 shares a seven-transmembrane-segment hydropathicity profile with other G protein-coupled receptors. Northern blot analyses identified a 2.7-kilobase transcript of the VIP receptor in Nalm 6 and HT-29 cells as well as in tissues from rat brain, colon, heart, lung, kidney, spleen, and small intestine. COS-6 cells transfected with GPRN1 bound 125I-labeled VIP specifically with a dissociation constant (Kd) of 2.5 nM. VIP--and less effectively secretin, peptide histidine isoleucine (PHI), and glucagon competitively displaced bound 125I-VIP from transfected COS-6 cells, with potencies in the order VIP greater than secretin = PHI much greater than glucagon. VIP stimulated adenylate cyclase activity in stably transfected Chinese hamster ovary K1 cells, inducing a 3-fold increase in the intracellular level of cAMP. When the antisense orientation of the VIP receptor clone was introduced into HT-29 cells, there was a 50% suppression of the specific binding of 125I-VIP and of the VIP-induced increase in cAMP level, relative to untransfected cells. The VIP receptor cloned exhibits less than or equal to 24% homology with other receptors in the same superfamily and thus represents a subset of G protein-coupled receptors for peptide ligands.

Amino Acid Sequence↗
PubMed1991

Affinity labeling of the membrane protein-binding component of human polymorphonuclear leukocyte receptors for leukotriene B4.

A radiolabeled N-(3-aminopropyl)-leukotriene B4 amide ([3H]LTB4-APA) analog of the potent leukocyte chemotactic factor leukotriene B4 (LTB4) binds to receptors for LTB4 in plasma membrane-enriched preparations from human blood polymorphonuclear leukocytes (PMNL) and intact PMNL with respective mean dissociation constants of 2.3 nM and 69 nM at 4 degrees C. The [3H]LTB4-APA bound to plasma membrane-enriched preparations from PMNL was covalently cross-linked to membrane proteins with disuccinimidyl suberate. Solubilization and resolution by SDS-PAGE of proteins from [3H]LTB4-APA-labeled PMNL membranes revealed predominant labeling of a 60-kDa protein. Labeling of the PMNL membrane protein was inhibited by LTB4 and its analogs at concentrations similar to those inhibiting the binding of [3H]LTB4 to its receptor, with an identical rank order of potency of LTB4 greater than 20-hydroxy-LTB4 greater than LTB4-APA = 5(S),12(R)-dihydroxy-eicosa-14-cis-6,8,10-trans-tetraenoic acid much greater than LTD4 = LTC4. GTP suppressed the labeling of the 60-kDa PMNL membrane protein to an extent consistent with the decrease in receptor affinity for LTB4 induced by GTP. The stereospecificity of the affinity cross-linking reaction and the regulation by GTP support the identification of an approximately 60-kDa protein as the binding component of the PMNL receptor for LTB4.

Affinity Labels↗
PubMed1991

Relative quantification of collagen mRNA in fibroblasts by a radioactive polymerase chain reaction technique.

A radioactive polymerase chain reaction (PCR) method has been developed for the relative quantification of the human alpha-2 chain of type I collagen [hu alpha-2(I)] in cells. cDNAs generated by reverse transcription from the total pool of cytoplasmic RNA serve as a template for polymerase chain reaction amplification of a hu alpha-2(I) cDNA primed by two sequence-specific synthetic oligonucleotides. The distinctive 390 bp hu alpha-2(I) cDNA and two Aval fragments of 220 and 170 bp are identified by agarose gel electrophoresis. alpha-32P-dCTP of defined specific activity is included in the PCR reaction and the 390 bp cDNA is excised from the electrophoresis gel to permit direct radioactive quantification of hu alpha-2(I) mRNA. The amount of hu alpha-2(I) mRNA expressed in as few as 111 fibroblasts was determined reliably. In contrast, the hu alpha-2(I) mRNA from at least 5 x 10(5) fibroblasts was required for detection by Northern blot analysis developed with the same cDNA probe radiolabelled with alpha-32P-dCTP by random priming. Human bronchoalveolar lavage (BAL) fluids of six patients with fibrosing lung diseases stimulated the level of expression of hu alpha-2(I) mRNA in cultured human fibroblasts as determined by this technique. The radioactive PCR method thus quantifies hu alpha-2(I) mRNA in fibroblasts with sufficient sensitivity to study fibroblast activation in vitro and detect fibroblast stimuli in human clinical samples.

Autoradiography↗
PubMed1991

Chemotactic activity and receptor binding of neutrophil attractant/activation protein-1 (NAP-1) and structurally related host defense cytokines: interaction of NAP-2 with the NAP-1 receptor.

Neutrophil attractant/activation protein-1 (NAP-1) has sequence similarity to platelet factor-4 (PF-4) and to NAP-2 (a truncated from of connective tissue activating protein-III [CTAP-III(des 1-15)]. We compared chemotactic activity for neutrophils of these related proteins. We also included for comparison CTAP-III, CTAP-III(des 1-13), the C-terminal dodecapeptide of PF-4 [PF-4(59-70)], and C5a. Chemotactic potency (EC50) was highest for NAP-1 and C5a. Although chemotactic efficacy (peak percentage of neutrophils migrating) was comparable for C5a, NAP-1, and NAP-2, the NAP-2 response occurred only at concentrations 100-fold higher than the NAP-1 EC50 of 10(8) M. Data for the CTAP-III proteins confirmed that CTAP-III is not an attractant and that chemotactic activity appears as a result of cleavage of residues at the N-terminus to make CTAP-III(des 1-13) or NAP-2 [CTAP-III(des 1-15)]. Chemotactic activity of PF-4 was low and variable, with no significant response by neutrophils from six of nine subjects. In contrast, PF-4(59-70) regularly induced high chemotactic responses, although the EC50 of 1.6 x 10(5)M was 1,000-fold greater than that of NAP-1. The binding of fluoresceinated NAP-1 to neutrophils was inhibited by unlabeled NAP-1 or NAP-2 but not by PF-4 or PF-4 (59-70). This suggests that NAP-2 interacts with the neutrophil NAP-1 receptor. Despite the low chemotactic potency of NAP-2, it is a potential attractant at sites of injury because of the relatively large amounts of the parent CTAP-III released from platelets, as indicated by a serum concentration of approximately 10(-6) M.

Chemotaxis, Leukocyte↗
PubMed1991

Polymerase chain reaction amplification of messages for growth factors in cells from human bronchoalveolar lavage fluids.

Genetic messages for polypeptide growth factors were assessed in human alveolar macrophages, obtained by bronchoalveolar lavage (BAL) from normal subjects (N = 3) and from patients with pneumonia (N = 3), pulmonary lymphoma (N = 3), and idiopathic pulmonary fibrosis (N = 3). Complementary DNAs (cDNAs) were prepared by reverse transcription of the RNA extracted from alveolar macrophages before and after culture on a plastic surface. The cDNAs encoding 10 different growth factors were amplified for electrophoretic analysis by polymerase chain reaction with a pair of 3' and 5' primers specific for each factor. Alveolar macrophages from all normal subjects and patients expressed the messages for interleukin-1 beta and transforming growth factor-beta. Alveolar macrophages from some normal subjects also contained message for insulin-like growth factor-1. Alveolar macrophages from six of nine patients with lung diseases also expressed messages for one or more additional growth factors, including epidermal growth factor, transforming growth factor-alpha, interleukin-1 alpha, and platelet-derived growth factor. The polymerase chain reaction technique thus permits determination of the profile of growth factors contributed to pulmonary reactions by alveolar macrophages, which may be important in pulmonary healing and fibrosis.

Bronchoalveolar Lavage Fluid↗
PubMed1991

Down-regulation of receptor antigen in leukotriene B4-induced chemotactic deactivation of human polymorphonuclear leucocytes.

Pretreatment of suspensions of human polymorphonuclear leucocytes (PMNL) with leukotriene B4 (LTB4) induces chemotactic deactivation, characterized by diminished expression of high-affinity LTB4 receptors and selectively decreased chemotactic responsiveness of the PMNL to LTB4. Rabbit anti-idiotypic antibodies (a-Id) to mouse monoclonal anti-LTB4, which bind to 60,000-80,000 molecular weight (MW) membrane protein of PMNL receptors for LTB4 in Western blots and block binding of [3H]LTB4 to high-affinity receptors of PMNL, detected a reduction in LTB4 receptor antigen during chemotactic deactivation. Loss of high-affinity receptors for LTB4 from the surface of PMNL deactivated by incubation with 10 nM LTB4 was significant after 1 min and after 20 min reached a mean maximum of 82% and 61%, respectively, as assessed by binding of [3H]LTB4 and a-Id. Inhibitors of PMNL proteases did not prevent the deactivation-induced decreases in surface receptors for LTB4. Disruption of deactivated PMNL and solubilization of membrane proteins failed to expose intracellular LTB4 receptors. Incubation of membranes isolated from PMNL with 100 nM LTB4 resulted in a loss of LTB4 receptors similar to that observed in intact PMNL. Changes in LTB4 receptor protein structure or membrane localization, rather than endocytosis or proteolysis, thus appear to explain the rapidly decreased expression of LTB4 receptors, which results from stimulus-specific deactivation.

Antibodies, Anti-Idiotypic↗
PubMed1991

Isolation and partial characterization of the structures of fibroblast activating factor-related proteins from U937 cells.

Human cultured monocyte-like tumour cells of the U937 histiocyte derived line were stimulated with phorbol myristate acetate, and generated and released an 18,000 MW polypeptide fibroblast-activating factor (FAF). Based on recognition by an antiserum to a synthetic peptide representing the 17 amino-terminal amino acids of FAF, two proteins of 32,000 and 35,000 MW were identified in extracts of U937 cells. Purification of the intracellular FAF-related proteins to homogeneity allowed the generation and amino acid sequencing of nine tryptic fragments of 4-11 amino acids. Neither of the intracellular FAF-related proteins exhibited the fibroblast proliferation-stimulating activity of FAF, suggesting that they are biosynthetic precursors analogous to the inactive propeptides of interleukin-1 beta and tumour necrosis factor-alpha.

Amino Acid Sequence↗
PubMed1990

Mediation of prolonged increases in nasal mucosal blood flow by calcitonin gene-related peptide (CGRP).

Calcitonin gene-related peptide (CGRP) is a vasoactive and smooth muscle contractile neuropeptide, which is released during human nasal allergic reactions. Mean values for nasal blood flow (NBF), quantified in each nostril by a laser Doppler technique, varied no more than +/- 25% of a mean initial reading in normal human subjects over 4 hr in a controlled environment. Intranasal administration of multiple doses of CGRP to 17 normal subjects stimulated significant mean increases in NBF at 30-1000 ng of CGRP, with maximal mean rises of 86-130% after 100 and 300 ng of CGRP. Delivery of incremental doses of CGRP every 30 min for 2 hr evoked repetitive responses without tachyphylaxis. A single vasoactive dose of CGRP, which achieved nasal fluid concentrations similar to those elicited by antigen challenge, elicited the sensation of nasal obstruction in most subjects at 1 hr and for up to 4 hr after CGRP, without elevating nasal fluid concentrations of histamine. CGRP thus is a potent histamine-independent mediator of allergic reactions.

Administration, Intranasal↗
PubMed1990

Suppression by ingested eicosapentaenoic acid of the increases in nasal mucosal blood flow and eosinophilia of ryegrass-allergic reactions.

Nasal mucosal blood flow, assessed by a laser Doppler probe technique, and the concentration of eosinophils in nasal secretions were quantified during challenge of one nostril with ryegrass-pollen antigen and the other nostril with diluent alone in seven patients with ryegrass-allergic rhinitis. The identical studies were repeated after an 8-week course of 3.5 gm/day of eicosapentaenoic acid (EPA). Ryegrass antigen evoked mean rises in nasal blood flow of 30% to 100% after 10 and 30 minutes that were significant, relative to prechallenge levels and to levels after diluent challenge, both before and after EPA. Antigen-induced increases in nasal blood flow were significantly less after than before EPA at 10 minutes, and at 180 minutes increases were significant only before EPA. In ryegrass-allergic patients with rhinitis who did not take EPA between the two studies, the increases in blood flow after antigen challenge were the same on both occasions. Similarly, the nasal eosinophilia elicited by antigen was significant at 180 minutes only before EPA. Both a composite index of signs and symptoms and the constituent variables, reflecting the clinical response to antigen challenge, were unaffected by EPA. The suppression by EPA of responses of nasal blood flow and nasal eosinophils to antigen challenge supports a role for fatty acid and phospholipid mediators in allergic rhinitis, but the clinical assessment did not provide evidence for any symptomatic benefit from EPA.

Adult↗