Regulation of human skin mast cell histamine release by PDE inhibitors

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Feng, Q. He, Y. Fan, J. Mi, L. Guo, H. Hong, H. Li" "autores" => array:7 [ 0 => array:2 [ "nombre" => "S." "apellidos" => "Feng" ] 1 => array:2 [ "nombre" => "Q." "apellidos" => "He" ] 2 => array:2 [ "nombre" => "Y." "apellidos" => "Fan" ] 3 => array:2 [ "nombre" => "J." "apellidos" => "Mi" ] 4 => array:2 [ "nombre" => "L." "apellidos" => "Guo" ] 5 => array:2 [ "nombre" => "H." "apellidos" => "Hong" ] 6 => array:2 [ "nombre" => "H." "apellidos" => "Li" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054614000251?idApp=UINPBA00004N" "url" => "/03010546/0000004300000001/v1_201501250943/S0301054614000251/v1_201501250943/en/main.assets" ] "itemAnterior" => array:17 [ "pii" => "S0301054613002310" "issn" => "03010546" "doi" => "10.1016/j.aller.2013.07.004" "estado" => "S300" "fechaPublicacion" => "2015-01-01" "aid" => "549" "copyright" => "SEICAP" "documento" => "article" "subdocumento" => "fla" "cita" => "Allergol Immunopathol (Madr). 2015;43:32-6" "abierto" => array:3 [ "ES" => false "ES2" => false "LATM" => false ] "gratuito" => false "lecturas" => array:2 [ "total" => 802 "formatos" => array:3 [ "EPUB" => 13 "HTML" => 513 "PDF" => 276 ] ] "en" => array:12 [ "idiomaDefecto" => true "cabecera" => "Original article" "titulo" => "Can meteorological factors forecast asthma exacerbation in a paediatric population?" "tienePdf" => "en" "tieneTextoCompleto" => "en" "tieneResumen" => "en" "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "32" "paginaFinal" => "36" ] ] "contieneResumen" => array:1 [ "en" => true ] "contieneTextoCompleto" => array:1 [ "en" => true ] "contienePdf" => array:1 [ "en" => true ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1113 "Ancho" => 1660 "Tamanyo" => 79950 ] ] "descripcion" => array:1 [ "en" => "Seasonality of asthma exacerbations in the paediatric emergency department." ] ] ] "autores" => array:1 [ 0 => array:2 [ "autoresLista" => "D. Hervás, J.F. Utrera, J. Hervás-Masip, J.A. Hervás, L. García-Marcos" "autores" => array:5 [ 0 => array:2 [ "nombre" => "D." "apellidos" => "Hervás" ] 1 => array:2 [ "nombre" => "J.F." "apellidos" => "Utrera" ] 2 => array:2 [ "nombre" => "J." "apellidos" => "Hervás-Masip" ] 3 => array:2 [ "nombre" => "J.A." "apellidos" => "Hervás" ] 4 => array:2 [ "nombre" => "L." "apellidos" => "García-Marcos" ] ] ] ] ] "idiomaDefecto" => "en" "EPUB" => "https://multimedia.elsevier.es/PublicationsMultimediaV1/item/epub/S0301054613002310?idApp=UINPBA00004N" "url" => "/03010546/0000004300000001/v1_201501250943/S0301054613002310/v1_201501250943/en/main.assets" ] "en" => array:18 [ "idiomaDefecto" => true "cabecera" => "Original Article" "titulo" => "Regulation of human skin mast cell histamine release by PDE inhibitors" "tieneTextoCompleto" => true "paginas" => array:1 [ 0 => array:2 [ "paginaInicial" => "37" "paginaFinal" => "41" ] ] "autores" => array:1 [ 0 => array:4 [ "autoresLista" => "N. Eskandari, R. Bastan, P.T. Peachell" "autores" => array:3 [ 0 => array:4 [ "nombre" => "N." "apellidos" => "Eskandari" "email" => array:1 [ 0 => "neskandari@med.mui.ac.ir" ] "referencia" => array:2 [ 0 => array:2 [ "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:2 [ "etiqueta" => "*" "identificador" => "cor0005" ] ] ] 1 => array:4 [ "nombre" => "R." "apellidos" => "Bastan" "email" => array:1 [ 0 => "rbastan@gmail.com" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "b" "identificador" => "aff0010" ] ] ] 2 => array:4 [ "nombre" => "P.T." "apellidos" => "Peachell" "email" => array:1 [ 0 => "P.T.Peachell@sheffield.ac.uk" ] "referencia" => array:1 [ 0 => array:2 [ "etiqueta" => "c" "identificador" => "aff0015" ] ] ] ] "afiliaciones" => array:3 [ 0 => array:3 [ "entidad" => "Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Postal code: 81744-176, Isfahan, Iran" "etiqueta" => "a" "identificador" => "aff0005" ] 1 => array:3 [ "entidad" => "Department of Immunpharmacology, Faculty of Medicine, Karaj, Iran" "etiqueta" => "b" "identificador" => "aff0010" ] 2 => array:3 [ "entidad" => "Department of Immunology, Faculty of Medicine, Sheffield University of Medical Sciences, Yorkshire, UK" "etiqueta" => "c" "identificador" => "aff0015" ] ] "correspondencia" => array:1 [ 0 => array:3 [ "identificador" => "cor0005" "etiqueta" => "⁎" "correspondencia" => "Corresponding author." ] ] ] ] "resumenGrafico" => array:2 [ "original" => 0 "multimedia" => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 924 "Ancho" => 1560 "Tamanyo" => 66738 ] ] "descripcion" => array:1 [ "en" => "Effect of 8-Br-cAMP (■) and 8-Br-cGMP (▴) on human skin mast cells (HSMC). Cells were incubated for 20min with either 8-bromo-cAMP or 8-Br-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25min. Results are presented as the percent inhibition of the control histamine release which was 15±3%. Values are means±SEM, n=6." ] ] ] "textoCompleto" => "IntroductionAt least eleven different classes of phosphodiesterase (PDE) have been identified based on structural and functional criteria.<a class="elsevierStyleCrossRefs" href="#bib0005">1–4</a> Enzymes within this family are found in most pro-inflammatory and immune cells where they are important regulators of the metabolism of cyclic nucleotides. Of the eleven PDE isoenzymes, types 3 and 4 hydrolyse cAMP and type 5, cGMP. The activity of PDEs can be modulated by inhibitors. These inhibitors can be divided into distinct categories: (a) classical non-specific inhibitors of PDE activity such as theophylline, and 3-isobutyl-1-methylxanthine (IBMX), and (b) selective PDE inhibitors such as 8-methoxy-methyl-IBMX (8-Me-IBMX; PDE 1 inhibitor), siguazodan (PDE 3 inhibitor), rolipram and denbufylline (PDE 4 inhibitors) and zaprinast (PDE 5 inhibitor). Pharmacological investigations, using selective and non-selective inhibitors of PDE isoenzymes, have shown that PDE type 3 and 4 inhibitors are more effective than PDE type 1, 2 and 5 inhibitors in inflammatory cells (8–9).Since PDEs hydrolyse cAMP and cGMP, the mechanism by which PDE inhibitors act is to elevate levels of these cyclic nucleotides. Some PDEs hydrolyse cAMP (PDE 4) preferentially whereas others are cGMP-selective (PDE 5). Cyclic AMP and cyclic GMP, as intracellular messengers, play vital roles regulating inflammatory cell activity (9). Since cAMP and cGMP are almost unable to penetrate intact cell membranes, several hundred cyclic nucleotide analogues, with hydrophobic substituent, have been synthesised and have been widely used to elucidate the functional role of cAMP and cGMP signal cascades in biological systems.<a class="elsevierStyleCrossRefs" href="#bib0025">5,6</a> Four of these, namely, 2′-O-dibutyrylguanosine 3′,5′-cyclic monophosphate (Bu2-cGMP), 2′-O-dibutyryladenosine 3′,5′-cyclic monophosphate (Bu2-cAMP), 8-bromo-cyclic 3′,5′-adenosine monophosphate (8-Br-cAMP) and 8-bromo-cyclic 3′,5′-guanosine monophosphate (8-Br-cGMP) have received major attention and are widely used as tools for testing the role of cAMP/cGMP and protein kinase A (PKA)/protein kinase G (PKG) in biological systems because cAMP and cGMP exert their physiological functions almost solely via activation of PKA and PKG, respectively.The main aim of the present study was to characterise the type and role of PDEs regulating human skin mast cells by using selective and non-selective PDE inhibitors. Secondly, a comparison of the effects of these inhibitors on related cell types, human lung mast cells and basophiles, was also undertaken. Thirdly, an investigation of the effects of non-hydrolysable analogues of cAMP and cGMP on skin mast cells was also undertaken to determine the role of cyclic nucleotides in the regulation of skin mast cells.Materials and methodsBuffersPhosphate-buffered saline (PBS) was employed in these studies. PBS contained (mM): NaCl 137; Na2HPO4·12H2O 8; KCl 2.7; KH2PO4 1.5. PBS–bovine serum albumin (BSA) was PBS which additionally contained: CaCl2·2H2O 1mM; MgCl2·6H2O 1mM; glucose 5.6mM; BSA 1mgml−1; DNase 15¿gml−1. PBS–human serum albumin (HSA) was PBS additionally supplemented with: CaCl2·2H2O 1mM; MgCl2·6H2O 1mM; glucose 5.6mM; HSA 30¿gml−1. The pH of all PBS buffers was titrated to 7.3.Preparation of inhibitors and stimuliPhosphodiesterase (PDE) inhibitors were made up as follows: rolipram, IBMX, Org 30029, 8-methoxy-methyl-IBMX, denbufylline and Ro-201724 were made up as 10−1M and 10−2M stock solutions in 10% DMSO. Siguazodan and theophylline were made up as 1mM and 10mM stock solution, respectively, in +PBS. Zaprinast was prepared as a 100mM stock solution in 0.1M NaOH. Cyclic nucleotide analogues were made up as follows: 8-bromo-c-AMP, 8-bromo-c-GMP, Bu2-cAMP and Bu2-cGMP were prepared as 100mM stock solutions made in +PBS daily. The stimulus used in mediator release experiments was polyclonal goat anti-human IgE which was prepared according to the manufacturer's instructions. The lyophilised powder was reconstituted in 2ml of ultra-pure H2O.Isolation of basophiles, human lung mast cells and skin mast cellsMixed leucocyte preparations were obtained from whole blood (from healthy and different volunteers) by dextran sedimentation. Briefly 50ml of venous blood was mixed with 12.5ml of 6% dextran and 5ml of 100mM EDTA, and then allowed to sediment for 90min at room temperature. The upper buffy coat layer was removed; cells were recovered by centrifugation (400×g, 8min) and washed twice with PBS. These mixed cell preparations were used in the histamine release experiments. Mast cells were isolated from normal human lung tissue by a modification of the method described by Ali and Pearce (1985). Macroscopically normal tissue from lung resections of patients was obtained with the approval of the Local Research Ethics Committee. The tissue was chopped vigorously for 10min with scissors in a small volume of PBS buffer and then washed over a nylon mesh (100¿m pore size; Incamesh, Warrington, UK) with 0.5–1l of PBS buffer to remove lung macrophages. The tissue was reconstituted in PBS–BSA (10ml per gram of tissue) containing collagenase Ia (0.1mgml−1 of PBS–BSA) and agitated by using a water-driven magnetic stirrer immersed in a water bath set at 37°C. The supernatant was separated from the tissue by filtration over nylon mesh. The collagenase-treated tissue was then reconstituted in a small volume of PBS–BSA buffer and disrupted mechanically with a syringe. The disrupted tissue was then washed over nylon gauze with PBS–BSA. The pooled filtrates were sedimented (400×g, room temperature, 8min), the supernatant discarded and the pellets reconstituted in PBS–BSA (100ml). The pellet was washed twice more. Mast cells were visualised by microscopy using an alcian blue stain. Of the total cells, 3–13% were mast cells. This method generated 2–9×105 mast cells per gram of tissue. Mast cells prepared in this manner were used in mediator release experiments.MethodsHistamine release experiments were performed in +PBS buffer and assessed in duplicate. Cell suspensions of about 2×104 human mast cells were used per sample. Cells were incubated for either 10 or 15min with the desired concentration of drug in a total volume of, usually, 300¿l and then 30¿l anti-human IgE (1/300 or 1/3000 for mast cells or basophiles, respectively) was added. Histamine release was allowed to proceed for 25 (mast cells) or 45 (basophiles) min at 37°C (Ennis, 1991). Histamine release reactions were terminated by adding 750¿l PBS to all samples. The samples were centrifuged (400×g, 4min, RT) and the histamine present in the cell supernatants was measured by a modification of the automated fluorometric technique of<a class="elsevierStyleCrossRef" href="#bib0035">7</a>. Total histamine content of the cells was determined by lysing aliquots of the cells with 1.8% (v/v) perchloric acid. Cells incubated in buffer alone served as a measure of spontaneous histamine release. Histamine release was calculated as a percentage of the total histamine content after subtracting spontaneous histamine release.Data analysisMaximal responses (Emax) and potencies (pEC50) were determined by non-linear regression analysis (GraphPad Prism, version 3.0a). To determine whether there was any difference in the responses after treatments with drugs, repeated measures analysis of variance was performed.MaterialsThe following were purchased from the sources indicated; anti-human IgE, aprotinin, BSA, collagenase, dimethyl sulphoxide, DNase, Dowex AG-50W, Dowex AG1-X8, dithiothreitol, HSA, leupeptin, Percoll, phenyl methyl sulphonyl fluoride, soybean trypsin inhibitor, Tween 20, Triton X-100 (all Sigma, Poole, UK); EDTA, calcium chloride and magnesium chloride (BDH, Poole, UK); goat polyclonal anti-human IgE, human serum albumin (HSA), dimethyl suphoxide (DMSO), sodium metabisulphite, rolipram, zaprinast, 3-isobutyl-1-methylxanthine (IBMX), 8-methoxy-methyl-IBMX, theophylline, isoprenaline, N6,2′-O-dibutyryladenosine 3′,5′-cyclic monophosphate (Bu2-cAMP) and N6,2′-O-dibutyrylguanosine 3′,5′-cyclic monophosphate (Bu2-cGMP); Org 30029 (gift from Dr CD Nicholson, Organon, UK); denbufylline (Beecham Pharmaceuticals, UK); 8-Bromo-cAMP, 8-Bromo-cGMP, and siguazodan (Tocris Cookson Ltd., UK); ethanol (BDH, Pool, UK); Ro 20-1724 (Research Biochemicals Incorporated, USA).ResultsEffects of PDE inhibitors on IgE-mediated histamine release from skin mast cellsThe effects of non-selective PDE inhibitors, theophylline and IBMX, on histamine release from human skin mast cells (HSMC) were investigated. Cells were incubated with increasing concentrations of PDE inhibitors for 15min before challenge with anti-IgE (1:300). The PDE inhibitors attenuated IgE-mediated histamine release in a dose-dependent manner and to a statistically significant (p<0.05) extent. IBMX was a more potent inhibitor than theophylline of IgE-mediated histamine release in HSMC. EC50 values for the IBMX and theophylline inhibition of histamine release from HSMC were 0.1mM and 1mM, respectively (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>).<elsevierMultimedia ident="fig0005"></elsevierMultimedia>Effects of cAMP and cGMP analogues on mast cellsThe non-selective inhibitors, IBMX and theophylline, are likely to exert effects on HSMC by elevating cAMP and/or GMP. To investigate the roles of cAMP and cGMP on HSMC the effects of cell-permeant, non-hydrolysable analogues of cAMP and cGMP were studied. The effects of Bu2-cAMP, Bu2-cGMP, 8-Br-cAMP and 8-Br-cGMP on IgE-mediated histamine release from HSMC were studied. Cells were pre-treated for 15min in the presence of increasing concentrations (3×10−5–3×10−3M) of these cyclic nucleotide analogues. Then the cells were triggered with an optimal releasing concentration of anti-IgE (1:300) for a further 25min for the release of histamine. Neither 8-Br-cAMP nor 8-Br-cGMP (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>) displayed any inhibitory activity on histamine release from HSMC. In further studies the effects of Bu2-cAMP and Bu2-cyclic GMP were investigated. The data show (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>) both Bu2-cAMP and Bu2-cGMP inhibit histamine release in a dose-dependent manner with maximal inhibitory effects of 63±9% and 53±5%, respectively at 3mM.<elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia>For comparative reasons, the effects of these analogues in HLMC were investigated. Neither 8-Br-cAMP nor 8-Br-cGMP had an inhibitory effect on HLMC (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>), whereas Bu2-cAMP, but not Bu2-cGMP, inhibited the stimulated release of histamine from HLMC (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>).<elsevierMultimedia ident="fig0020"></elsevierMultimedia><elsevierMultimedia ident="fig0025"></elsevierMultimedia>Effects of selective PDE inhibitors on HSMC, HLMC, and basophilesBecause PDE 4 is the predominant isoform in inflammatory cells,<a class="elsevierStyleCrossRefs" href="#bib0040">8,9</a> the effects of the PDE 4-selective inhibitor, rolipram, on IgE-mediated histamine release from HSMC were studied. Cells were pre-treated for 15min with increasing concentrations of rolipram (10−10–10−5M) or DMSO (vehicle) then triggered with an optimal releasing concentration of anti-IgE (1:300) for a further 25min for histamine release. Rolipram (and DMSO over the same equivalent vehicle concentration range of rolipram) failed to inhibit histamine release from HSMC (data not shown).The inability of rolipram to stabilise HSMC responses prompted us to investigate the effects of a wide range of selective PDE inhibitors on the IgE-mediated histamine release from HSMC. Cells were pre-treated for 15min with these agents and then challenged with an optimal releasing concentration of anti IgE (1:300) for a further 25min for the release of histamine. The data show (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>) that all the selective PDE inhibitor compounds (10−5M) were ineffective whereas the non-selective PDE inhibitor, theophylline (10−3M), inhibited histamine release from HSMC (74±4% inhibition; p<0.05).<elsevierMultimedia ident="tbl0005"></elsevierMultimedia>For comparative purposes, the effects of these same compounds on the IgE-mediated release of histamine from both HLMC and basophiles were investigated (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). None of the selective PDE inhibitors had any effect on histamine release from HLMC whereas, in basophiles, compounds with activity at PDE 4 (rolipram, denbufylline, Ro-2017, Org 30029) were effective inhibitors of histamine release.DiscussionThe results of the present study show that theophylline and IBMX act as effective inhibitors of IgE-mediated histamine release from HSMC. These results suggest that inhibition of PDE can lead to the reduction of the secretory response in skin mast cells. Inhibition of PDE would be expected to cause increases in cyclic nucleotides. Alternative studies indicate that treatment of HLMC with IBMX causes intracellular elevations in cyclic AMP<a class="elsevierStyleCrossRef" href="#bib0050">10</a> although, the effect that non-selective PDE inhibitors have on cGMP content in human mast cells is not known.In order to gain a better understanding of the roles of cGMP and cAMP in HSMC, we investigated the effects of cell-permeant and non-hydrolysable analogues of these cyclic nucleotides. Neither 8-Br-cAMP nor 8-Br-cGMP had any effects in HSMC. Indeed, to determine whether this was a finding restricted to HSMC, the effects of these analogues on HLMC were investigated. These analogues were ineffective on these cells and further studies from our group<a class="elsevierStyleCrossRef" href="#bib0055">11</a> have shown that 8-bromo derivatives are ineffective in basophiles. This could be because the 8-bromo derivatives are not very cell-permeant. Therefore, we decided to look at alternative analogues Bu2-cAMP and Bu2-cGMP. Both analogues were effective inhibitors of histamine release from HSMC, suggesting that both cAMP and cGMP may inhibit the responses of HSMC. However, caution is needed with this interpretation as dibutyryl compounds can be cleaved intracellulary to dibutyrate which is known to affect cell function. However, arguing against an effect of butyrate on HSMC is the finding that Bu2-cAMP, and not Bu2-cGMP, inhibited histamine release from HLMC.In further studies the effects of selective PDE inhibitors on HSMC were investigated. None of the inhibitors tested had any effect on the IgE-mediated release of histamine. Interestingly, therefore, HSMC behave much like HLMC which are also insensitive to PDE 4 inhibitors (8). These findings suggest that the PDE that regulates HSMC and HLMC function is something other than PDE 1, 3, 4, or 5. Mast cells, therefore, differ from the vast majority of inflammatory cells which are known to be regulated by PDE 4<a class="elsevierStyleCrossRefs" href="#bib0040">8,9</a>. This includes the human basophile, which the present study has shown to be particularly sensitive to inhibitors of PDE4, confirming previous studies<a class="elsevierStyleCrossRefs" href="#bib0045">9,11</a>. It is possible that the methods used to prepare HSMC or that the maturation state of the isolated mast cells which were obtained from the foreskins of neonates and infants could influence the expression of PDEs. This issue could be resolved by isolating mast cells from adult skin. However, that HLMC are also refractory to PDE4 inhibitors makes it probable that human mast cells, in general, do not express PDE4.The emphasis of the present study was to characterise PDE isoforms in HSMC. The data suggest that unlike most inflammatory cells, PDE4-selective inhibitors are ineffective stabilisers of HSMC activity. In this regard HSMC are similar to HLMC. However, due to the functional heterogeneity displayed by mast cells the possibility exists that the responses to PDE inhibitors of HSMC and HLMC may not necessarily reflect those of alternative subsets of mast cells. In summary, the present work has shown that although those PDEs appear to regulate HSMC, the nature of this PDE (s) remains uncertain.Ethical disclosuresConfidentiality of dataThe authors declare that no patient data appear in this article.Right to privacy and informed consentThe authors have obtained the informed consent of the patients and/or subjects mentioned in the article. The author for correspondence is in possession of this document.Protection of human and animal subjectsThe authors declare that the procedures followed were in accordance with the regulations of the responsible Clinical Research Ethics Committee and in accordance with those of the World Medical Association and the Helsinki Declaration.Conflict of interestThe authors have no conflict of interest to declare." "textoCompletoSecciones" => array:1 [ "secciones" => array:9 [ 0 => array:2 [ "identificador" => "xres414311" "titulo" => array:5 [ 0 => "Abstract" 1 => "Background" 2 => "Methods" 3 => "Results" 4 => "Conclusion" ] ] 1 => array:2 [ "identificador" => "xpalclavsec389951" "titulo" => "Keywords" ] 2 => array:2 [ "identificador" => "sec0005" "titulo" => "Introduction" ] 3 => array:3 [ "identificador" => "sec0010" "titulo" => "Materials and methods" "secciones" => array:6 [ 0 => array:2 [ "identificador" => "sec0015" "titulo" => "Buffers" ] 1 => array:2 [ "identificador" => "sec0020" "titulo" => "Preparation of inhibitors and stimuli" ] 2 => array:2 [ "identificador" => "sec0025" "titulo" => "Isolation of basophiles, human lung mast cells and skin mast cells" ] 3 => array:2 [ "identificador" => "sec0030" "titulo" => "Methods" ] 4 => array:2 [ "identificador" => "sec0035" "titulo" => "Data analysis" ] 5 => array:2 [ "identificador" => "sec0040" "titulo" => "Materials" ] ] ] 4 => array:3 [ "identificador" => "sec0045" "titulo" => "Results" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0050" "titulo" => "Effects of PDE inhibitors on IgE-mediated histamine release from skin mast cells" ] 1 => array:2 [ "identificador" => "sec0055" "titulo" => "Effects of cAMP and cGMP analogues on mast cells" ] 2 => array:2 [ "identificador" => "sec0060" "titulo" => "Effects of selective PDE inhibitors on HSMC, HLMC, and basophiles" ] ] ] 5 => array:2 [ "identificador" => "sec0065" "titulo" => "Discussion" ] 6 => array:3 [ "identificador" => "sec0070" "titulo" => "Ethical disclosures" "secciones" => array:3 [ 0 => array:2 [ "identificador" => "sec0075" "titulo" => "Confidentiality of data" ] 1 => array:2 [ "identificador" => "sec0080" "titulo" => "Right to privacy and informed consent" ] 2 => array:2 [ "identificador" => "sec0085" "titulo" => "Protection of human and animal subjects" ] ] ] 7 => array:2 [ "identificador" => "sec0090" "titulo" => "Conflict of interest" ] 8 => array:1 [ "titulo" => "References" ] ] ] "pdfFichero" => "main.pdf" "tienePdf" => true "fechaRecibido" => "2013-06-11" "fechaAceptado" => "2013-07-20" "PalabrasClave" => array:1 [ "en" => array:1 [ 0 => array:4 [ "clase" => "keyword" "titulo" => "Keywords" "identificador" => "xpalclavsec389951" "palabras" => array:3 [ 0 => "Mast cell" 1 => "Histamine" 2 => "PDE" ] ] ] ] "tieneResumen" => true "resumen" => array:1 [ "en" => array:2 [ "titulo" => "Abstract" "resumen" => "BackgroundMast cell and basophiles are thought to be central to inflammation that has an allergic basis as allergens activate these cells in an IgE-dependent manner to generate mediators such as histamine, eicosanoids and cytokines. Phosphodiesterase (PDE) is known to exist as multiple molecular forms of enzyme that metabolise the second messengers. Studies of our own have shown that, of a variety of isoform-selective drugs, the PDE4-selective inhibitors, such as rolipram, attenuate the IgE-mediated release of histamine from human basophiles but not from human lung mast cells (HLMC).The main aim of the present study was to characterise the type and role of PDEs regulating human skin mast cells by using selective and non-selective PDE inhibitors. MethodsCells were pre-treated for 15min with these agents and then challenged with an optimal releasing concentration of anti IgE (1:300) for a further 25min for the release of histamine. ResultsThe data show that all the selective PDE-inhibitor compounds (10−5M) were ineffective whereas the non-selective PDE inhibitor, theophylline (10−3M), inhibited histamine release from HSMC (74±4% inhibition; p<0.05). None of the selective PDE inhibitors had any effect on histamine release from HLMC whereas, in basophiles, compounds with activity at PDE 4 (rolipram, denbufylline, Ro-2017, Org 30029) were effective inhibitors of histamine release. ConclusionThe data suggest that unlike most inflammatory cells, PDE-selective inhibitors are ineffective stabilisers of HSMC activity which is similar to HLMC." ] ] "multimedia" => array:6 [ 0 => array:7 [ "identificador" => "fig0005" "etiqueta" => "Figure 1" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr1.jpeg" "Alto" => 1115 "Ancho" => 1513 "Tamanyo" => 60100 ] ] "descripcion" => array:1 [ "en" => "Effect of non-selective PDE inhibitors on histamine release from human skin mast cells (HSMC). The effect of either IBMX (■) or theophylline (●) on the release of histamine from HSMC was determined. Cells were incubated for 15min with a PDE inhibitor and then challenged with an optimal releasing concentration of anti-IgE (1:300) for a further 25min. Results are expressed as the percent inhibition of the control release, which was 19±4%. Statistically significant (p<0.01) levels of inhibition are indicated by asterisks. Values are means±SEM, n=5." ] ] 1 => array:7 [ "identificador" => "fig0010" "etiqueta" => "Figure 2" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr2.jpeg" "Alto" => 924 "Ancho" => 1560 "Tamanyo" => 66738 ] ] "descripcion" => array:1 [ "en" => "Effect of 8-Br-cAMP (■) and 8-Br-cGMP (▴) on human skin mast cells (HSMC). Cells were incubated for 20min with either 8-bromo-cAMP or 8-Br-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25min. Results are presented as the percent inhibition of the control histamine release which was 15±3%. Values are means±SEM, n=6." ] ] 2 => array:7 [ "identificador" => "fig0015" "etiqueta" => "Figure 3" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr3.jpeg" "Alto" => 999 "Ancho" => 1507 "Tamanyo" => 62357 ] ] "descripcion" => array:1 [ "en" => "Effect of Bu2-cAMP (■) and Bu2-cGMP (▴) on human skin mast cells (HSMC). Cells were incubated for 20min with either Bu2-cAMP or Bu2-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25min. Results are presented as the percent inhibition of the control histamine release which was 18±2%. Values are means±SEM, n=10." ] ] 3 => array:7 [ "identificador" => "fig0020" "etiqueta" => "Figure 4" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr4.jpeg" "Alto" => 1008 "Ancho" => 1572 "Tamanyo" => 60203 ] ] "descripcion" => array:1 [ "en" => "Effect of 8-Br-cAMP (■) and 8-Br-cGMP (▴) on human lung mast cells (HLMC). Cells were incubated for 20min with either 8-Br-cAMP or 8-Br-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25min. Results are presented as the percent inhibition of the control histamine release which was 38±3%. Values are means±SEM, n=3." ] ] 4 => array:7 [ "identificador" => "fig0025" "etiqueta" => "Figure 5" "tipo" => "MULTIMEDIAFIGURA" "mostrarFloat" => true "mostrarDisplay" => false "figura" => array:1 [ 0 => array:4 [ "imagen" => "gr5.jpeg" "Alto" => 955 "Ancho" => 1583 "Tamanyo" => 55891 ] ] "descripcion" => array:1 [ "en" => "Effect of Bu2-cAMP (■) and Bu2-cGMP (△) on human lung mast cells (HLMC). Cells were incubated for 20min with either Bu2-cAMP or Bu2-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25min. Results are presented as the percent inhibition of the control histamine release which was 43±8%. Values are means±SEM, n=4." ] ] 5 => array:7 [ "identificador" => "tbl0005" "etiqueta" => "Table 1" "tipo" => "MULTIMEDIATABLA" "mostrarFloat" => true "mostrarDisplay" => false "tabla" => array:1 [ "tablatextoimagen" => array:1 [ 0 => array:2 [ "tabla" => array:1 [ 0 => """ <table border="0" frame="\n \t\t\t\t\tvoid\n \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="table-head " align="left" valign="top" scope="col">PDE inhibitor class \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">% Inhibition</th></tr><tr title="table-row"><th class="td" title="table-head " align="" valign="top" scope="col" style="border-bottom: 2px solid black"> \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">HSMC \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">HLMC \t\t\t\t\t\t\n \t\t\t\t</th><th class="td" title="table-head " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Basophiles \t\t\t\t\t\t\n \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">8-Me-IBMX (1) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">16.0±4.9 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.4±4.1 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">6.9±2.7 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Siguazodan (3) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">20.3±4.9 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.5±2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">5.0±5.5 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Org 30029 (3/4) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">0.9±7.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.3±3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">45.9±1.8* \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Rolipram (4) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">23.3±6.2 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">10.2±4.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">67.5±2.2* \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Denbufylline (4) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">20.1±5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">11.5±2.7 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">49.3±1.0* \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Ro-2017 (4) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">17.5±6 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.5±5.3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">40.9±2.4* \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Zaprinast (5) \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">13.3±5.5 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">9.5±3 \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">10.6±8.8 \t\t\t\t\t\t\n \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry " align="left" valign="top">Theophylline N.S. \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">74.2±4.2* \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">71.3±4.1* \t\t\t\t\t\t\n \t\t\t\t</td><td class="td" title="table-entry " align="char" valign="top">77.8±2.1* \t\t\t\t\t\t\n \t\t\t\t</td></tr></tbody></table> """ ] "imagenFichero" => array:1 [ 0 => "xTab646885.png" ] ] ] ] "descripcion" => array:1 [ "en" => "Effects of selective PDE inhibitors on histamine release from human skin mast cells (HSMC), human lung mast cells (HLMC), and human peripheral blood basophils." ] ] ] "bibliografia" => array:2 [ "titulo" => "References" "seccion" => array:1 [ 0 => array:2 [ "identificador" => "bibs0005" "bibliografiaReferencia" => array:11 [ 0 => array:3 [ "identificador" => "bib0005" "etiqueta" => "1" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Primary sequence of cyclic nucleotide phosphodiesterase isozymes and the design of selective inhibitors" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "J.A. Beavo" 1 => "D.H. Reifsnyder" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1016/0165-6147(90)90066-H" "Revista" => array:6 [ "tituloSerie" => "Trends Pharmacol Sci" "fecha" => "1990" "volumen" => "11" "paginaInicial" => "150" "paginaFinal" => "155" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/2159198" "web" => "Medline" ] ] ] ] ] ] ] ] 1 => array:3 [ "identificador" => "bib0010" "etiqueta" => "2" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Multiple cyclic nucleotide phosphodiesterases" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "J.A. Beavo" 1 => "M. Conti" 2 => "R.J. Heaslip" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Mol Pharmacol" "fecha" => "1994" "volumen" => "46" "paginaInicial" => "399" "paginaFinal" => "405" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/7935318" "web" => "Medline" ] ] ] ] ] ] ] ] 2 => array:3 [ "identificador" => "bib0015" "etiqueta" => "3" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Inhibitors of cyclic nucleotide phosphodiesterase isoenzymes--their potential utility in the therapy of asthma" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:2 [ 0 => "C.D. Nicholson" 1 => "M. Shahid" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Pulm Pharmacol" "fecha" => "1994" "volumen" => "7" "paginaInicial" => "1" "paginaFinal" => "17" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/8003849" "web" => "Medline" ] ] ] ] ] ] ] ] 3 => array:3 [ "identificador" => "bib0020" "etiqueta" => "4" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Isolation and characterization of PDE9A: a novel human cGMP-specific phosphodiesterase" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "D.A. Fisher" 1 => "J.F. Smith" 2 => "J.S. Pillar" 3 => "S.H. St Denis" 4 => "J.B. Cheng" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "J Biol Chem" "fecha" => "1998" "volumen" => "273" "paginaInicial" => "15559" "paginaFinal" => "15564" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/9624146" "web" => "Medline" ] ] ] ] ] ] ] ] 4 => array:3 [ "identificador" => "bib0025" "etiqueta" => "5" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Trial treatment of psoriasis by intralesional injections of AMPc, 8-thio-dibutyryl AMPc and theophylline" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:5 [ 0 => "F. Posternak" 1 => "T. Posternak" 2 => "M. Orusco" 3 => "G. Cehovic" 4 => "P. Laugier" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Ann Dermatol Syphiligr (Paris)" "fecha" => "1976" "volumen" => "103" "numero" => "5-6" "paginaInicial" => "640" "paginaFinal" => "644" ] ] ] ] ] ] 5 => array:3 [ "identificador" => "bib0030" "etiqueta" => "6" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "2’ Derivatives of guanosine and inosine cyclic 3’,5’-phosphates. Synthesis, enzymic activity, and the effect of 8-substituents" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:6 [ 0 => "J.P. Miller" 1 => "K.H. Boswell" 2 => "A.M. Mian" 3 => "R.B. Meyer Jr." 4 => "R.K. Robins" 5 => "T.A. Khwaja" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Biochemistry" "fecha" => "1976" "volumen" => "15" "paginaInicial" => "217" "paginaFinal" => "223" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/174713" "web" => "Medline" ] ] ] ] ] ] ] ] 6 => array:3 [ "identificador" => "bib0035" "etiqueta" => "7" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "An automated continuous-flow system for the extraction and fluorometric analysis of histamine" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:1 [ 0 => "R.P. Siraganian" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Anal Biochem" "fecha" => "1974" "volumen" => "57" "paginaInicial" => "383" "paginaFinal" => "394" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/4819732" "web" => "Medline" ] ] ] ] ] ] ] ] 7 => array:3 [ "identificador" => "bib0040" "etiqueta" => "8" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Evidence that cyclic AMP phosphodiesterase inhibitors suppress TNF alpha generation from human monocytes by interacting with a ‘low-affinity’ phosphodiesterase 4 conformer" "autores" => array:1 [ 0 => array:2 [ "etal" => true "autores" => array:6 [ 0 => "J.E. Souness" 1 => "M. Griffin" 2 => "C. Maslen" 3 => "K. Ebsworth" 4 => "L.C. Scott" 5 => "K. Pollock" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "Br J Pharmacol" "fecha" => "1996" "volumen" => "118" "paginaInicial" => "649" "paginaFinal" => "658" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/8762090" "web" => "Medline" ] ] ] ] ] ] ] ] 8 => array:3 [ "identificador" => "bib0045" "etiqueta" => "9" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Effects of phosphodiesterase inhibitors on human lung mast cell and basophil function" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "M.C. Weston" 1 => "N. Anderson" 2 => "P.T. Peachell" ] ] ] ] ] "host" => array:1 [ 0 => array:2 [ "doi" => "10.1038/sj.bjp.0701115" "Revista" => array:6 [ "tituloSerie" => "Br J Pharmacol" "fecha" => "1997" "volumen" => "121" "paginaInicial" => "287" "paginaFinal" => "295" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/9154339" "web" => "Medline" ] ] ] ] ] ] ] ] 9 => array:3 [ "identificador" => "bib0050" "etiqueta" => "10" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Differential regulation of human basophil and lung mast cell function by adenosine" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "P.T. Peachell" 1 => "L.M. Lichtenstein" 2 => "R.P. Schleimer" ] ] ] ] ] "host" => array:1 [ 0 => array:1 [ "Revista" => array:6 [ "tituloSerie" => "J Pharmacol Exp Ther" "fecha" => "1991" "volumen" => "256" "paginaInicial" => "717" "paginaFinal" => "726" "link" => array:1 [ 0 => array:2 [ "url" => "https://www.ncbi.nlm.nih.gov/pubmed/1704436" "web" => "Medline" ] ] ] ] ] ] ] ] 10 => array:3 [ "identificador" => "bib0055" "etiqueta" => "11" "referencia" => array:1 [ 0 => array:2 [ "contribucion" => array:1 [ 0 => array:2 [ "titulo" => "Effects of phosphodiesterase inhibitors on interleukin-4 and interleukin-13 generation from human basophils" "autores" => array:1 [ 0 => array:2 [ "etal" => false "autores" => array:3 [ 0 => "N. Eskandari" 1 => "T. Wickramasinghe" 2 => "P.T. 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Original Article

N. Eskandaria,

Corresponding author

neskandari@med.mui.ac.ir

Corresponding author.

, R. Bastanb, P.T. Peachellc

a Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Postal code: 81744-176, Isfahan, Iran

b Department of Immunpharmacology, Faculty of Medicine, Karaj, Iran

c Department of Immunology, Faculty of Medicine, Sheffield University of Medical Sciences, Yorkshire, UK

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These inhibitors can be divided into distinct categories: (a) classical non-specific inhibitors of PDE activity such as theophylline, and 3-isobutyl-1-methylxanthine (IBMX), and (b) selective PDE inhibitors such as 8-methoxy-methyl-IBMX (8-Me-IBMX; PDE 1 inhibitor), siguazodan (PDE 3 inhibitor), rolipram and denbufylline (PDE 4 inhibitors) and zaprinast (PDE 5 inhibitor). Pharmacological investigations, using selective and non-selective inhibitors of PDE isoenzymes, have shown that PDE type 3 and 4 inhibitors are more effective than PDE type 1, 2 and 5 inhibitors in inflammatory cells (8&#8211;9).</p><p id="par0010" class="elsevierStylePara elsevierViewall">Since PDEs hydrolyse cAMP and cGMP, the mechanism by which PDE inhibitors act is to elevate levels of these cyclic nucleotides. Some PDEs hydrolyse cAMP (PDE 4) preferentially whereas others are cGMP-selective (PDE 5). Cyclic AMP and cyclic GMP, as intracellular messengers, play vital roles regulating inflammatory cell activity (9). Since cAMP and cGMP are almost unable to penetrate intact cell membranes, several hundred cyclic nucleotide analogues, with hydrophobic substituent, have been synthesised and have been widely used to elucidate the functional role of cAMP and cGMP signal cascades in biological systems.<a class="elsevierStyleCrossRefs" href="#bib0025"><span class="elsevierStyleSup">5,6</span></a> Four of these, namely, 2&#8242;-O-dibutyrylguanosine 3&#8242;,5&#8242;-cyclic monophosphate (Bu<span class="elsevierStyleInf">2</span>-cGMP), 2&#8242;-O-dibutyryladenosine 3&#8242;,5&#8242;-cyclic monophosphate (Bu<span class="elsevierStyleInf">2</span>-cAMP), 8-bromo-cyclic 3&#8242;,5&#8242;-adenosine monophosphate (8-Br-cAMP) and 8-bromo-cyclic 3&#8242;,5&#8242;-guanosine monophosphate (8-Br-cGMP) have received major attention and are widely used as tools for testing the role of cAMP/cGMP and protein kinase A (PKA)/protein kinase G (PKG) in biological systems because cAMP and cGMP exert their physiological functions almost solely via activation of PKA and PKG, respectively.</p><p id="par0015" class="elsevierStylePara elsevierViewall">The main aim of the present study was to characterise the type and role of PDEs regulating human skin mast cells by using selective and non-selective PDE inhibitors. Secondly, a comparison of the effects of these inhibitors on related cell types, human lung mast cells and basophiles, was also undertaken. Thirdly, an investigation of the effects of non-hydrolysable analogues of cAMP and cGMP on skin mast cells was also undertaken to determine the role of cyclic nucleotides in the regulation of skin mast cells.</p></span><span id="sec0010" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0040">Materials and methods</span><span id="sec0015" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0045">Buffers</span><p id="par0020" class="elsevierStylePara elsevierViewall">Phosphate-buffered saline (PBS) was employed in these studies. PBS contained (mM): NaCl 137; Na<span class="elsevierStyleInf">2</span>HPO<span class="elsevierStyleInf">4</span>&middot;12H<span class="elsevierStyleInf">2</span>O 8; KCl 2.7; KH<span class="elsevierStyleInf">2</span>PO<span class="elsevierStyleInf">4</span> 1.5. PBS&#8211;bovine serum albumin (BSA) was PBS which additionally contained: CaCl<span class="elsevierStyleInf">2</span>&middot;2H<span class="elsevierStyleInf">2</span>O 1<span class="elsevierStyleHsp" style=""></span>mM; MgCl<span class="elsevierStyleInf">2</span>&middot;6H<span class="elsevierStyleInf">2</span>O 1<span class="elsevierStyleHsp" style=""></span>mM; glucose 5.6<span class="elsevierStyleHsp" style=""></span>mM; BSA 1<span class="elsevierStyleHsp" style=""></span>mg<span class="elsevierStyleHsp" style=""></span>ml<span class="elsevierStyleSup">&#8722;1</span>; DNase 15<span class="elsevierStyleHsp" style=""></span>¿g<span class="elsevierStyleHsp" style=""></span>ml<span class="elsevierStyleSup">&#8722;1</span>. PBS&#8211;human serum albumin (HSA) was PBS additionally supplemented with: CaCl<span class="elsevierStyleInf">2</span><span class="elsevierStyleBold">&middot;</span>2H<span class="elsevierStyleInf">2</span>O 1<span class="elsevierStyleHsp" style=""></span>mM; MgCl<span class="elsevierStyleInf">2</span>&middot;6H<span class="elsevierStyleInf">2</span>O 1<span class="elsevierStyleHsp" style=""></span>mM; glucose 5.6<span class="elsevierStyleHsp" style=""></span>mM; HSA 30<span class="elsevierStyleHsp" style=""></span>¿g<span class="elsevierStyleHsp" style=""></span>ml<span class="elsevierStyleSup">&#8722;1</span>. The pH of all PBS buffers was titrated to 7.3.</p></span><span id="sec0020" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0050">Preparation of inhibitors and stimuli</span><p id="par0025" class="elsevierStylePara elsevierViewall">Phosphodiesterase (PDE) inhibitors were made up as follows: rolipram, IBMX, Org 30029, 8-methoxy-methyl-IBMX, denbufylline and Ro-201724 were made up as 10<span class="elsevierStyleSup">&#8722;1</span><span class="elsevierStyleHsp" style=""></span>M and 10<span class="elsevierStyleSup">&#8722;2</span><span class="elsevierStyleHsp" style=""></span>M stock solutions in 10% DMSO. Siguazodan and theophylline were made up as 1<span class="elsevierStyleHsp" style=""></span>mM and 10<span class="elsevierStyleHsp" style=""></span>mM stock solution, respectively, in +PBS. Zaprinast was prepared as a 100<span class="elsevierStyleHsp" style=""></span>mM stock solution in 0.1<span class="elsevierStyleHsp" style=""></span>M NaOH. Cyclic nucleotide analogues were made up as follows: 8-bromo-c-AMP, 8-bromo-c-GMP, Bu<span class="elsevierStyleInf">2</span>-cAMP and Bu<span class="elsevierStyleInf">2</span>-cGMP were prepared as 100<span class="elsevierStyleHsp" style=""></span>mM stock solutions made in +PBS daily. The stimulus used in mediator release experiments was polyclonal goat anti-human IgE which was prepared according to the manufacturer&#39;s instructions. The lyophilised powder was reconstituted in 2<span class="elsevierStyleHsp" style=""></span>ml of ultra-pure H<span class="elsevierStyleInf">2</span>O.</p></span><span id="sec0025" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0055">Isolation of basophiles, human lung mast cells and skin mast cells</span><p id="par0030" class="elsevierStylePara elsevierViewall">Mixed leucocyte preparations were obtained from whole blood (from healthy and different volunteers) by dextran sedimentation. Briefly 50<span class="elsevierStyleHsp" style=""></span>ml of venous blood was mixed with 12.5<span class="elsevierStyleHsp" style=""></span>ml of 6% dextran and 5<span class="elsevierStyleHsp" style=""></span>ml of 100<span class="elsevierStyleHsp" style=""></span>mM EDTA, and then allowed to sediment for 90<span class="elsevierStyleHsp" style=""></span>min at room temperature. The upper buffy coat layer was removed; cells were recovered by centrifugation (400<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span>, 8<span class="elsevierStyleHsp" style=""></span>min) and washed twice with PBS. These mixed cell preparations were used in the histamine release experiments. Mast cells were isolated from normal human lung tissue by a modification of the method described by Ali and Pearce (1985). Macroscopically normal tissue from lung resections of patients was obtained with the approval of the Local Research Ethics Committee. The tissue was chopped vigorously for 10<span class="elsevierStyleHsp" style=""></span>min with scissors in a small volume of PBS buffer and then washed over a nylon mesh (100<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">¿</span>m pore size; Incamesh, Warrington, UK) with 0.5&#8211;1<span class="elsevierStyleHsp" style=""></span>l of PBS buffer to remove lung macrophages. The tissue was reconstituted in PBS&#8211;BSA (10<span class="elsevierStyleHsp" style=""></span>ml per gram of tissue) containing collagenase Ia (0.1<span class="elsevierStyleHsp" style=""></span>mg<span class="elsevierStyleHsp" style=""></span>ml<span class="elsevierStyleSup">&#8722;1</span> of PBS&#8211;BSA) and agitated by using a water-driven magnetic stirrer immersed in a water bath set at 37<span class="elsevierStyleHsp" style=""></span>&deg;C. The supernatant was separated from the tissue by filtration over nylon mesh. The collagenase-treated tissue was then reconstituted in a small volume of PBS&#8211;BSA buffer and disrupted mechanically with a syringe. The disrupted tissue was then washed over nylon gauze with PBS&#8211;BSA. The pooled filtrates were sedimented (400<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span>, room temperature, 8<span class="elsevierStyleHsp" style=""></span>min), the supernatant discarded and the pellets reconstituted in PBS&#8211;BSA (100<span class="elsevierStyleHsp" style=""></span>ml). The pellet was washed twice more. Mast cells were visualised by microscopy using an alcian blue stain. Of the total cells, 3&#8211;13% were mast cells. This method generated 2&#8211;9<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">5</span> mast cells per gram of tissue. Mast cells prepared in this manner were used in mediator release experiments.</p></span><span id="sec0030" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0060">Methods</span><p id="par0035" class="elsevierStylePara elsevierViewall">Histamine release experiments were performed in +PBS buffer and assessed in duplicate. Cell suspensions of about 2<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">4</span> human mast cells were used per sample. Cells were incubated for either 10 or 15<span class="elsevierStyleHsp" style=""></span>min with the desired concentration of drug in a total volume of, usually, 300<span class="elsevierStyleHsp" style=""></span>¿l and then 30<span class="elsevierStyleHsp" style=""></span>¿l anti-human IgE (1/300 or 1/3000 for mast cells or basophiles, respectively) was added. Histamine release was allowed to proceed for 25 (mast cells) or 45 (basophiles) min at 37<span class="elsevierStyleHsp" style=""></span>&deg;C (Ennis, 1991). Histamine release reactions were terminated by adding 750<span class="elsevierStyleHsp" style=""></span>¿l PBS to all samples. The samples were centrifuged (400<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span><span class="elsevierStyleItalic">g</span>, 4<span class="elsevierStyleHsp" style=""></span>min, RT) and the histamine present in the cell supernatants was measured by a modification of the automated fluorometric technique of<a class="elsevierStyleCrossRef" href="#bib0035"><span class="elsevierStyleSup">7</span></a>. Total histamine content of the cells was determined by lysing aliquots of the cells with 1.8% (v/v) perchloric acid. Cells incubated in buffer alone served as a measure of spontaneous histamine release. Histamine release was calculated as a percentage of the total histamine content after subtracting spontaneous histamine release.</p></span><span id="sec0035" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0065">Data analysis</span><p id="par0040" class="elsevierStylePara elsevierViewall">Maximal responses (<span class="elsevierStyleItalic">E</span><span class="elsevierStyleInf">max</span>) and potencies (pEC<span class="elsevierStyleInf">50</span>) were determined by non-linear regression analysis (GraphPad Prism, version 3.0a). To determine whether there was any difference in the responses after treatments with drugs, repeated measures analysis of variance was performed.</p></span><span id="sec0040" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0070">Materials</span><p id="par0045" class="elsevierStylePara elsevierViewall">The following were purchased from the sources indicated; anti-human IgE, aprotinin, BSA, collagenase, dimethyl sulphoxide, DNase, Dowex AG-50W, Dowex AG1-X8, dithiothreitol, HSA, leupeptin, Percoll, phenyl methyl sulphonyl fluoride, soybean trypsin inhibitor, Tween 20, Triton X-100 (all Sigma, Poole, UK); EDTA, calcium chloride and magnesium chloride (BDH, Poole, UK); goat polyclonal anti-human IgE, human serum albumin (HSA), dimethyl suphoxide (DMSO), sodium metabisulphite, rolipram, zaprinast, 3-isobutyl-1-methylxanthine (IBMX), 8-methoxy-methyl-IBMX, theophylline, isoprenaline, N<span class="elsevierStyleSup">6</span>,2&#8242;-O-dibutyryladenosine 3&#8242;,5&#8242;-cyclic monophosphate (Bu<span class="elsevierStyleInf">2</span>-cAMP) and N<span class="elsevierStyleSup">6</span>,2&#8242;-O-dibutyrylguanosine 3&#8242;,5&#8242;-cyclic monophosphate (Bu<span class="elsevierStyleInf">2</span>-cGMP); Org 30029 (gift from Dr CD Nicholson, Organon, UK); denbufylline (Beecham Pharmaceuticals, UK); 8-Bromo-cAMP, 8-Bromo-cGMP, and siguazodan (Tocris Cookson Ltd., UK); ethanol (BDH, Pool, UK); Ro 20-1724 (Research Biochemicals Incorporated, USA).</p></span></span><span id="sec0045" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0075">Results</span><span id="sec0050" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0080">Effects of PDE inhibitors on IgE-mediated histamine release from skin mast cells</span><p id="par0050" class="elsevierStylePara elsevierViewall">The effects of non-selective PDE inhibitors, theophylline and IBMX, on histamine release from human skin mast cells (HSMC) were investigated. Cells were incubated with increasing concentrations of PDE inhibitors for 15<span class="elsevierStyleHsp" style=""></span>min before challenge with anti-IgE (1:300). The PDE inhibitors attenuated IgE-mediated histamine release in a dose-dependent manner and to a statistically significant (<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0.05) extent. IBMX was a more potent inhibitor than theophylline of IgE-mediated histamine release in HSMC. EC<span class="elsevierStyleInf">50</span> values for the IBMX and theophylline inhibition of histamine release from HSMC were 0.1<span class="elsevierStyleHsp" style=""></span>mM and 1<span class="elsevierStyleHsp" style=""></span>mM, respectively (<a class="elsevierStyleCrossRef" href="#fig0005">Fig. 1</a>).</p><elsevierMultimedia ident="fig0005"></elsevierMultimedia></span><span id="sec0055" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0085">Effects of cAMP and cGMP analogues on mast cells</span><p id="par0055" class="elsevierStylePara elsevierViewall">The non-selective inhibitors, IBMX and theophylline, are likely to exert effects on HSMC by elevating cAMP and/or GMP. To investigate the roles of cAMP and cGMP on HSMC the effects of cell-permeant, non-hydrolysable analogues of cAMP and cGMP were studied. The effects of Bu<span class="elsevierStyleInf">2</span>-cAMP, Bu<span class="elsevierStyleInf">2</span>-cGMP, 8-Br-cAMP and 8-Br-cGMP on IgE-mediated histamine release from HSMC were studied. Cells were pre-treated for 15<span class="elsevierStyleHsp" style=""></span>min in the presence of increasing concentrations (3<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">&#8722;5</span>&#8211;3<span class="elsevierStyleHsp" style=""></span>&times;<span class="elsevierStyleHsp" style=""></span>10<span class="elsevierStyleSup">&#8722;3</span><span class="elsevierStyleHsp" style=""></span>M) of these cyclic nucleotide analogues. Then the cells were triggered with an optimal releasing concentration of anti-IgE (1:300) for a further 25<span class="elsevierStyleHsp" style=""></span>min for the release of histamine. Neither 8-Br-cAMP nor 8-Br-cGMP (<a class="elsevierStyleCrossRef" href="#fig0010">Fig. 2</a>) displayed any inhibitory activity on histamine release from HSMC. In further studies the effects of Bu<span class="elsevierStyleInf">2</span>-cAMP and Bu<span class="elsevierStyleInf">2</span>-cyclic GMP were investigated. The data show (<a class="elsevierStyleCrossRef" href="#fig0015">Fig. 3</a>) both Bu<span class="elsevierStyleInf">2</span>-cAMP and Bu<span class="elsevierStyleInf">2</span>-cGMP inhibit histamine release in a dose-dependent manner with maximal inhibitory effects of 63<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>9% and 53<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>5%, respectively at 3<span class="elsevierStyleHsp" style=""></span>mM.</p><elsevierMultimedia ident="fig0010"></elsevierMultimedia><elsevierMultimedia ident="fig0015"></elsevierMultimedia><p id="par0060" class="elsevierStylePara elsevierViewall">For comparative reasons, the effects of these analogues in HLMC were investigated. Neither 8-Br-cAMP nor 8-Br-cGMP had an inhibitory effect on HLMC (<a class="elsevierStyleCrossRef" href="#fig0020">Fig. 4</a>), whereas Bu<span class="elsevierStyleInf">2</span>-cAMP, but not Bu<span class="elsevierStyleInf">2</span>-cGMP, inhibited the stimulated release of histamine from HLMC (<a class="elsevierStyleCrossRef" href="#fig0025">Fig. 5</a>).</p><elsevierMultimedia ident="fig0020"></elsevierMultimedia><elsevierMultimedia ident="fig0025"></elsevierMultimedia></span><span id="sec0060" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0090">Effects of selective PDE inhibitors on HSMC, HLMC, and basophiles</span><p id="par0065" class="elsevierStylePara elsevierViewall">Because PDE 4 is the predominant isoform in inflammatory cells,<a class="elsevierStyleCrossRefs" href="#bib0040"><span class="elsevierStyleSup">8,9</span></a> the effects of the PDE 4-selective inhibitor, rolipram, on IgE-mediated histamine release from HSMC were studied. Cells were pre-treated for 15<span class="elsevierStyleHsp" style=""></span>min with increasing concentrations of rolipram (10<span class="elsevierStyleSup">&#8722;10</span>&#8211;10<span class="elsevierStyleSup">&#8722;5</span><span class="elsevierStyleHsp" style=""></span>M) or DMSO (vehicle) then triggered with an optimal releasing concentration of anti-IgE (1:300) for a further 25<span class="elsevierStyleHsp" style=""></span>min for histamine release. Rolipram (and DMSO over the same equivalent vehicle concentration range of rolipram) failed to inhibit histamine release from HSMC (data not shown).</p><p id="par0070" class="elsevierStylePara elsevierViewall">The inability of rolipram to stabilise HSMC responses prompted us to investigate the effects of a wide range of selective PDE inhibitors on the IgE-mediated histamine release from HSMC. Cells were pre-treated for 15<span class="elsevierStyleHsp" style=""></span>min with these agents and then challenged with an optimal releasing concentration of anti IgE (1:300) for a further 25<span class="elsevierStyleHsp" style=""></span>min for the release of histamine. The data show (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>) that all the selective PDE inhibitor compounds (10<span class="elsevierStyleSup">&#8722;5</span><span class="elsevierStyleHsp" style=""></span>M) were ineffective whereas the non-selective PDE inhibitor, theophylline (10<span class="elsevierStyleSup">&#8722;3</span><span class="elsevierStyleHsp" style=""></span>M), inhibited histamine release from HSMC (74<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4% inhibition; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0.05).</p><elsevierMultimedia ident="tbl0005"></elsevierMultimedia><p id="par0075" class="elsevierStylePara elsevierViewall">For comparative purposes, the effects of these same compounds on the IgE-mediated release of histamine from both HLMC and basophiles were investigated (<a class="elsevierStyleCrossRef" href="#tbl0005">Table 1</a>). None of the selective PDE inhibitors had any effect on histamine release from HLMC whereas, in basophiles, compounds with activity at PDE 4 (rolipram, denbufylline, Ro-2017, Org 30029) were effective inhibitors of histamine release.</p></span></span><span id="sec0065" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0095">Discussion</span><p id="par0080" class="elsevierStylePara elsevierViewall">The results of the present study show that theophylline and IBMX act as effective inhibitors of IgE-mediated histamine release from HSMC. These results suggest that inhibition of PDE can lead to the reduction of the secretory response in skin mast cells. Inhibition of PDE would be expected to cause increases in cyclic nucleotides. Alternative studies indicate that treatment of HLMC with IBMX causes intracellular elevations in cyclic AMP<a class="elsevierStyleCrossRef" href="#bib0050"><span class="elsevierStyleSup">10</span></a> although, the effect that non-selective PDE inhibitors have on cGMP content in human mast cells is not known.</p><p id="par0085" class="elsevierStylePara elsevierViewall">In order to gain a better understanding of the roles of cGMP and cAMP in HSMC, we investigated the effects of cell-permeant and non-hydrolysable analogues of these cyclic nucleotides. Neither 8-Br-cAMP nor 8-Br-cGMP had any effects in HSMC. Indeed, to determine whether this was a finding restricted to HSMC, the effects of these analogues on HLMC were investigated. These analogues were ineffective on these cells and further studies from our group<a class="elsevierStyleCrossRef" href="#bib0055"><span class="elsevierStyleSup">11</span></a> have shown that 8-bromo derivatives are ineffective in basophiles. This could be because the 8-bromo derivatives are not very cell-permeant. Therefore, we decided to look at alternative analogues Bu<span class="elsevierStyleInf">2</span>-cAMP and Bu<span class="elsevierStyleInf">2</span>-cGMP. Both analogues were effective inhibitors of histamine release from HSMC, suggesting that both cAMP and cGMP may inhibit the responses of HSMC. However, caution is needed with this interpretation as dibutyryl compounds can be cleaved intracellulary to dibutyrate which is known to affect cell function. However, arguing against an effect of butyrate on HSMC is the finding that Bu<span class="elsevierStyleInf">2</span>-cAMP, and not Bu<span class="elsevierStyleInf">2</span>-cGMP, inhibited histamine release from HLMC.</p><p id="par0090" class="elsevierStylePara elsevierViewall">In further studies the effects of selective PDE inhibitors on HSMC were investigated. None of the inhibitors tested had any effect on the IgE-mediated release of histamine. Interestingly, therefore, HSMC behave much like HLMC which are also insensitive to PDE 4 inhibitors (8). These findings suggest that the PDE that regulates HSMC and HLMC function is something other than PDE 1, 3, 4, or 5. Mast cells, therefore, differ from the vast majority of inflammatory cells which are known to be regulated by PDE 4<a class="elsevierStyleCrossRefs" href="#bib0040"><span class="elsevierStyleSup">8,9</span></a>. This includes the human basophile, which the present study has shown to be particularly sensitive to inhibitors of PDE4, confirming previous studies<a class="elsevierStyleCrossRefs" href="#bib0045"><span class="elsevierStyleSup">9,11</span></a>. It is possible that the methods used to prepare HSMC or that the maturation state of the isolated mast cells which were obtained from the foreskins of neonates and infants could influence the expression of PDEs. This issue could be resolved by isolating mast cells from adult skin. However, that HLMC are also refractory to PDE4 inhibitors makes it probable that human mast cells, in general, do not express PDE4.</p><p id="par0095" class="elsevierStylePara elsevierViewall">The emphasis of the present study was to characterise PDE isoforms in HSMC. The data suggest that unlike most inflammatory cells, PDE4-selective inhibitors are ineffective stabilisers of HSMC activity. In this regard HSMC are similar to HLMC. However, due to the functional heterogeneity displayed by mast cells the possibility exists that the responses to PDE inhibitors of HSMC and HLMC may not necessarily reflect those of alternative subsets of mast cells. In summary, the present work has shown that although those PDEs appear to regulate HSMC, the nature of this PDE (s) remains uncertain.</p></span><span id="sec0070" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0100">Ethical disclosures</span><span id="sec0075" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0105">Confidentiality of data</span><p id="par0100" class="elsevierStylePara elsevierViewall">The authors declare that no patient data appear in this article.</p></span><span id="sec0080" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0110">Right to privacy and informed consent</span><p id="par0105" class="elsevierStylePara elsevierViewall">The authors have obtained the informed consent of the patients and/or subjects mentioned in the article. The author for correspondence is in possession of this document.</p></span><span id="sec0085" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0115">Protection of human and animal subjects</span><p id="par0110" class="elsevierStylePara elsevierViewall">The authors declare that the procedures followed were in accordance with the regulations of the responsible Clinical Research Ethics Committee and in accordance with those of the World Medical Association and the Helsinki Declaration.</p></span></span><span id="sec0090" class="elsevierStyleSection elsevierViewall"><span class="elsevierStyleSectionTitle" id="sect0120">Conflict of interest</span><p id="par0115" class="elsevierStylePara elsevierViewall">The authors have no conflict of interest to declare.</p></span></span>"
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        "titulo" => "Abstract"
        "resumen" => "<span class="elsevierStyleSectionTitle" id="sect0010">Background</span><p id="spar0005" class="elsevierStyleSimplePara elsevierViewall">Mast cell and basophiles are thought to be central to inflammation that has an allergic basis as allergens activate these cells in an IgE-dependent manner to generate mediators such as histamine, eicosanoids and cytokines. Phosphodiesterase (PDE) is known to exist as multiple molecular forms of enzyme that metabolise the second messengers. Studies of our own have shown that, of a variety of isoform-selective drugs, the PDE4-selective inhibitors, such as rolipram, attenuate the IgE-mediated release of histamine from human basophiles but not from human lung mast cells (HLMC).</p><p id="spar0010" class="elsevierStyleSimplePara elsevierViewall">The main aim of the present study was to characterise the type and role of PDEs regulating human skin mast cells by using selective and non-selective PDE inhibitors.</p> <span class="elsevierStyleSectionTitle" id="sect0015">Methods</span><p id="spar0015" class="elsevierStyleSimplePara elsevierViewall">Cells were pre-treated for 15<span class="elsevierStyleHsp" style=""></span>min with these agents and then challenged with an optimal releasing concentration of anti IgE (1:300) for a further 25<span class="elsevierStyleHsp" style=""></span>min for the release of histamine.</p> <span class="elsevierStyleSectionTitle" id="sect0020">Results</span><p id="spar0020" class="elsevierStyleSimplePara elsevierViewall">The data show that all the selective PDE-inhibitor compounds (10<span class="elsevierStyleSup">&#8722;5</span><span class="elsevierStyleHsp" style=""></span>M) were ineffective whereas the non-selective PDE inhibitor, theophylline (10<span class="elsevierStyleSup">&#8722;3</span><span class="elsevierStyleHsp" style=""></span>M), inhibited histamine release from HSMC (74<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4% inhibition; <span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0.05). None of the selective PDE inhibitors had any effect on histamine release from HLMC whereas, in basophiles, compounds with activity at PDE 4 (rolipram, denbufylline, Ro-2017, Org 30029) were effective inhibitors of histamine release.</p> <span class="elsevierStyleSectionTitle" id="sect0025">Conclusion</span><p id="spar0025" class="elsevierStyleSimplePara elsevierViewall">The data suggest that unlike most inflammatory cells, PDE-selective inhibitors are ineffective stabilisers of HSMC activity which is similar to HLMC.</p>"
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          "en" => "<p id="spar0030" class="elsevierStyleSimplePara elsevierViewall">Effect of non-selective PDE inhibitors on histamine release from human skin mast cells (HSMC). The effect of either IBMX (&#9632;) or theophylline (&#9679;) on the release of histamine from HSMC was determined. Cells were incubated for 15<span class="elsevierStyleHsp" style=""></span>min with a PDE inhibitor and then challenged with an optimal releasing concentration of anti-IgE (1:300) for a further 25<span class="elsevierStyleHsp" style=""></span>min. Results are expressed as the percent inhibition of the control release, which was 19<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4%. Statistically significant (<span class="elsevierStyleItalic">p</span><span class="elsevierStyleHsp" style=""></span>&#60;<span class="elsevierStyleHsp" style=""></span>0.01) levels of inhibition are indicated by asterisks. Values are means<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>SEM, <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>5.</p>"
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          "en" => "<p id="spar0035" class="elsevierStyleSimplePara elsevierViewall">Effect of 8-Br-cAMP (&#9632;) and 8-Br-cGMP (&#9652;) on human skin mast cells (HSMC). Cells were incubated for 20<span class="elsevierStyleHsp" style=""></span>min with either 8-bromo-cAMP or 8-Br-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25<span class="elsevierStyleHsp" style=""></span>min. Results are presented as the percent inhibition of the control histamine release which was 15<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>3%. Values are means<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>SEM, <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>6.</p>"
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        "tipo" => "MULTIMEDIAFIGURA"
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          0 => array:4 [
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        "descripcion" => array:1 [
          "en" => "<p id="spar0040" class="elsevierStyleSimplePara elsevierViewall">Effect of Bu<span class="elsevierStyleInf">2</span>-cAMP (&#9632;) and Bu<span class="elsevierStyleInf">2</span>-cGMP (&#9652;) on human skin mast cells (HSMC). Cells were incubated for 20<span class="elsevierStyleHsp" style=""></span>min with either Bu<span class="elsevierStyleInf">2</span>-cAMP or Bu<span class="elsevierStyleInf">2</span>-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25<span class="elsevierStyleHsp" style=""></span>min. Results are presented as the percent inhibition of the control histamine release which was 18<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2%. Values are means<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>SEM, <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>10.</p>"
        ]
      ]
      3 => array:7 [
        "identificador" => "fig0020"
        "etiqueta" => "Figure 4"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
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          0 => array:4 [
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0045" class="elsevierStyleSimplePara elsevierViewall">Effect of 8-Br-cAMP (&#9632;) and 8-Br-cGMP (&#9652;) on human lung mast cells (HLMC). Cells were incubated for 20<span class="elsevierStyleHsp" style=""></span>min with either 8-Br-cAMP or 8-Br-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25<span class="elsevierStyleHsp" style=""></span>min. Results are presented as the percent inhibition of the control histamine release which was 38<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>3%. Values are means<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>SEM, n<span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>3.</p>"
        ]
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      4 => array:7 [
        "identificador" => "fig0025"
        "etiqueta" => "Figure 5"
        "tipo" => "MULTIMEDIAFIGURA"
        "mostrarFloat" => true
        "mostrarDisplay" => false
        "figura" => array:1 [
          0 => array:4 [
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        ]
        "descripcion" => array:1 [
          "en" => "<p id="spar0050" class="elsevierStyleSimplePara elsevierViewall">Effect of Bu<span class="elsevierStyleInf">2</span>-cAMP (&#9632;) and Bu<span class="elsevierStyleInf">2</span>-cGMP (&#9651;) on human lung mast cells (HLMC). Cells were incubated for 20<span class="elsevierStyleHsp" style=""></span>min with either Bu<span class="elsevierStyleInf">2</span>-cAMP or Bu<span class="elsevierStyleInf">2</span>-cGMP before challenge with an optimal releasing concentration of anti-IgE (1:300). Histamine release was allowed to proceed for 25<span class="elsevierStyleHsp" style=""></span>min. Results are presented as the percent inhibition of the control histamine release which was 43<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>8%. Values are means<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>SEM, <span class="elsevierStyleItalic">n</span><span class="elsevierStyleHsp" style=""></span>=<span class="elsevierStyleHsp" style=""></span>4.</p>"
        ]
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                0 => """
                  <table border="0" frame="\n
                  \t\t\t\t\tvoid\n
                  \t\t\t\t" class=""><thead title="thead"><tr title="table-row"><th class="td" title="table-head  " align="left" valign="top" scope="col">PDE inhibitor class&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " colspan="3" align="center" valign="top" scope="col" style="border-bottom: 2px solid black">% Inhibition</th></tr><tr title="table-row"><th class="td" title="table-head  " align="" valign="top" scope="col" style="border-bottom: 2px solid black">&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">HSMC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">HLMC&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th><th class="td" title="table-head  " align="left" valign="top" scope="col" style="border-bottom: 2px solid black">Basophiles&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</th></tr></thead><tbody title="tbody"><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">8-Me-IBMX (1)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">16.0<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.9&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9.4<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.1&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">6.9<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2.7&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Siguazodan (3)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">20.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.9&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">5.0<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>5.5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Org 30029 (3/4)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">0.9<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>7.2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">45.9<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>1.8*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Rolipram (4)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">23.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>6.2&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">10.2<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">67.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2.2*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Denbufylline (4)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">20.1<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">11.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2.7&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">49.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>1.0*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Ro-2017 (4)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">17.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>6&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>5.3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">40.9<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2.4*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Zaprinast (5)&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">13.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>5.5&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">9.5<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>3&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">10.6<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>8.8&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr><tr title="table-row"><td class="td" title="table-entry  " align="left" valign="top">Theophylline N.S.&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">74.2<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.2*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">71.3<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>4.1*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td><td class="td" title="table-entry  " align="char" valign="top">77.8<span class="elsevierStyleHsp" style=""></span>&plusmn;<span class="elsevierStyleHsp" style=""></span>2.1*&nbsp;\t\t\t\t\t\t\n
                  \t\t\t\t</td></tr></tbody></table>
                  """
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        "descripcion" => array:1 [
          "en" => "<p id="spar0055" class="elsevierStyleSimplePara elsevierViewall">Effects of selective PDE inhibitors on histamine release from human skin mast cells (HSMC), human lung mast cells (HLMC), and human peripheral blood basophils.</p>"
        ]
      ]
    ]
    "bibliografia" => array:2 [
      "titulo" => "References"
      "seccion" => array:1 [
        0 => array:2 [
          "identificador" => "bibs0005"
          "bibliografiaReferencia" => array:11 [
            0 => array:3 [
              "identificador" => "bib0005"
              "etiqueta" => "1"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Primary sequence of cyclic nucleotide phosphodiesterase isozymes and the design of selective inhibitors"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "J.A. Beavo"
                            1 => "D.H. Reifsnyder"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:2 [
                      "doi" => "10.1016/0165-6147(90)90066-H"
                      "Revista" => array:6 [
                        "tituloSerie" => "Trends Pharmacol Sci"
                        "fecha" => "1990"
                        "volumen" => "11"
                        "paginaInicial" => "150"
                        "paginaFinal" => "155"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/2159198"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            1 => array:3 [
              "identificador" => "bib0010"
              "etiqueta" => "2"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Multiple cyclic nucleotide phosphodiesterases"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:3 [
                            0 => "J.A. Beavo"
                            1 => "M. Conti"
                            2 => "R.J. Heaslip"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Mol Pharmacol"
                        "fecha" => "1994"
                        "volumen" => "46"
                        "paginaInicial" => "399"
                        "paginaFinal" => "405"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/7935318"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            2 => array:3 [
              "identificador" => "bib0015"
              "etiqueta" => "3"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Inhibitors of cyclic nucleotide phosphodiesterase isoenzymes--their potential utility in the therapy of asthma"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:2 [
                            0 => "C.D. Nicholson"
                            1 => "M. Shahid"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Pulm Pharmacol"
                        "fecha" => "1994"
                        "volumen" => "7"
                        "paginaInicial" => "1"
                        "paginaFinal" => "17"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/8003849"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            3 => array:3 [
              "identificador" => "bib0020"
              "etiqueta" => "4"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Isolation and characterization of PDE9A: a novel human cGMP-specific phosphodiesterase"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "D.A. Fisher"
                            1 => "J.F. Smith"
                            2 => "J.S. Pillar"
                            3 => "S.H. St Denis"
                            4 => "J.B. Cheng"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "J Biol Chem"
                        "fecha" => "1998"
                        "volumen" => "273"
                        "paginaInicial" => "15559"
                        "paginaFinal" => "15564"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/9624146"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            4 => array:3 [
              "identificador" => "bib0025"
              "etiqueta" => "5"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Trial treatment of psoriasis by intralesional injections of AMPc, 8-thio-dibutyryl AMPc and theophylline"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:5 [
                            0 => "F. Posternak"
                            1 => "T. Posternak"
                            2 => "M. Orusco"
                            3 => "G. Cehovic"
                            4 => "P. Laugier"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Ann Dermatol Syphiligr (Paris)"
                        "fecha" => "1976"
                        "volumen" => "103"
                        "numero" => "5-6"
                        "paginaInicial" => "640"
                        "paginaFinal" => "644"
                      ]
                    ]
                  ]
                ]
              ]
            ]
            5 => array:3 [
              "identificador" => "bib0030"
              "etiqueta" => "6"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "2&#8217; Derivatives of guanosine and inosine cyclic 3&#8217;,5&#8217;-phosphates. Synthesis, enzymic activity, and the effect of 8-substituents"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:6 [
                            0 => "J.P. Miller"
                            1 => "K.H. Boswell"
                            2 => "A.M. Mian"
                            3 => "R.B. Meyer Jr."
                            4 => "R.K. Robins"
                            5 => "T.A. Khwaja"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Biochemistry"
                        "fecha" => "1976"
                        "volumen" => "15"
                        "paginaInicial" => "217"
                        "paginaFinal" => "223"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/174713"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            6 => array:3 [
              "identificador" => "bib0035"
              "etiqueta" => "7"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "An automated continuous-flow system for the extraction and fluorometric analysis of histamine"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => false
                          "autores" => array:1 [
                            0 => "R.P. Siraganian"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Anal Biochem"
                        "fecha" => "1974"
                        "volumen" => "57"
                        "paginaInicial" => "383"
                        "paginaFinal" => "394"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/4819732"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            7 => array:3 [
              "identificador" => "bib0040"
              "etiqueta" => "8"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
                    0 => array:2 [
                      "titulo" => "Evidence that cyclic AMP phosphodiesterase inhibitors suppress TNF alpha generation from human monocytes by interacting with a &#8216;low-affinity&#8217; phosphodiesterase 4 conformer"
                      "autores" => array:1 [
                        0 => array:2 [
                          "etal" => true
                          "autores" => array:6 [
                            0 => "J.E. Souness"
                            1 => "M. Griffin"
                            2 => "C. Maslen"
                            3 => "K. Ebsworth"
                            4 => "L.C. Scott"
                            5 => "K. Pollock"
                          ]
                        ]
                      ]
                    ]
                  ]
                  "host" => array:1 [
                    0 => array:1 [
                      "Revista" => array:6 [
                        "tituloSerie" => "Br J Pharmacol"
                        "fecha" => "1996"
                        "volumen" => "118"
                        "paginaInicial" => "649"
                        "paginaFinal" => "658"
                        "link" => array:1 [
                          0 => array:2 [
                            "url" => "https://www.ncbi.nlm.nih.gov/pubmed/8762090"
                            "web" => "Medline"
                          ]
                        ]
                      ]
                    ]
                  ]
                ]
              ]
            ]
            8 => array:3 [
              "identificador" => "bib0045"
              "etiqueta" => "9"
              "referencia" => array:1 [
                0 => array:2 [
                  "contribucion" => array:1 [
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Article information

ISSN: 03010546
Original language: English

DOI:10.1016/j.aller.2013.07.011

The statistics are updated each day

Year/Month	Html	Pdf	Total

2024 November	2	2	4
2024 October	26	8	34
2024 September	39	20	59
2024 August	37	15	52
2024 July	20	17	37
2024 June	28	9	37
2024 May	23	9	32
2024 April	29	16	45
2024 March	42	18	60
2024 February	46	9	55
2024 January	66	10	76
2023 December	52	17	69
2023 November	64	19	83
2023 October	86	20	106
2023 September	58	7	65
2023 August	42	12	54
2023 July	67	7	74
2023 June	63	14	77
2023 May	50	6	56
2023 April	39	7	46
2023 March	32	4	36
2023 February	25	7	32
2023 January	33	8	41
2022 December	39	4	43
2022 November	20	14	34
2022 October	19	10	29
2022 September	20	9	29
2022 August	23	10	33
2022 July	26	9	35
2022 June	22	6	28
2022 May	45	12	57
2022 April	51	13	64
2022 March	30	11	41
2022 February	16	3	19
2022 January	22	7	29
2021 December	14	10	24
2021 November	19	14	33
2021 October	29	14	43
2021 September	21	12	33
2021 August	17	4	21
2021 July	52	10	62
2021 June	17	23	40
2021 May	22	9	31
2021 April	38	12	50
2021 March	58	8	66
2021 February	45	13	58
2021 January	35	18	53
2020 December	4	4	8
2020 November	0	1	1
2020 October	1	1	2
2020 September	0	2	2
2020 July	0	1	1
2020 May	0	2	2
2020 April	0	1	1
2020 February	0	2	2
2019 October	0	1	1
2019 June	0	2	2
2019 May	0	9	9
2018 March	4	1	5
2018 February	20	1	21
2018 January	29	1	30
2017 December	15	2	17
2017 November	28	1	29
2017 October	17	2	19
2017 September	19	6	25
2017 August	31	6	37
2017 July	27	6	33
2017 June	30	18	48
2017 May	41	7	48
2017 April	29	5	34
2017 March	19	38	57
2017 February	37	7	44
2017 January	30	0	30
2016 December	20	3	23
2016 November	20	8	28
2016 October	53	12	65
2016 September	13	11	24
2016 August	27	4	31
2016 July	11	1	12
2016 June	18	14	32
2016 May	15	9	24
2016 April	20	10	30
2016 March	15	11	26
2016 February	18	14	32
2016 January	21	8	29
2015 October	0	1	1
2015 April	0	5	5
2015 March	4	4	8
2015 February	4	4	8

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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Allergologia et Immunopathologia is no longer published on Elsevier since the 2021 year.Transferred to Codon Publications

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