RESUMO
OBJECTIVES: Immunisation against preventable diseases as meningitis is crucial from a public health perspective to face challenges posed by these infections. Nurses hold a great responsibility for these programs, which highlights the importance of understanding their preferences and needs to improve the success of campaigns. This study aimed to investigate nurses' preferences regarding Meningococcus A, C, W, and Y (MenACWY) conjugate vaccines commercialised in Spain. STUDY DESIGN: A national-level discrete choice experiment (DCE) was conducted. METHODS: A literature review and a focus group informed the DCE design. Six attributes were included: pharmaceutical form, coadministration evidence, shelf-life, package contents, single-doses per package, and package volume. Conditional logit models quantified preferences and relative importance (RI). RESULTS: Thirty experienced primary care nurses participated in this study. Evidence of coadministration with other vaccines was the most important attribute (RI = 43.78%), followed by package size (RI = 22.17%), pharmaceutical form (RI = 19.07%), and package content (RI = 11.80%). There was a preference for evidence of coadministration with routine vaccines (odds ratio [OR] = 2.579, 95% confidence interval [95%CI] = 2.210-3.002), smaller volumes (OR = 1.494, 95%CI = 1.264-1.767), liquid formulations (OR = 1.283, 95%CI = 1.108-1.486) and package contents including only vial/s (OR = 1.283, 95%CI = 1.108-1.486). No statistical evidence was found for the remaining attributes. CONCLUSIONS: Evidence of coadministration with routine vaccines, easy-to-store packages, and fully liquid formulations were drivers of nurses' preferences regarding MenACWY conjugate vaccines. These findings provide valuable insights for decision-makers to optimize current campaigns.
Assuntos
Vacinas Meningocócicas , Neisseria meningitidis , Enfermeiras e Enfermeiros , Humanos , Espanha , Vacinas Conjugadas , Comportamento de Escolha , Preparações FarmacêuticasRESUMO
Respiratory syncytial virus (RSV) is a leading cause of acute lower respiratory infection (ALRI) in children, causing frequent outpatient visits and hospitalizations. Our study aimed to describe the clinical and direct economic burden of ALRI hospitalizations related to RSV in children in Spain and the characteristics of patients and their episodes. In this retrospective study, ALRI hospitalizations in children aged < 5 years for 2015-2018 were reviewed using anonymized administrative public hospital discharge data from Spain. Three case definitions were considered: (a) RSV-specific; (b) RSV-specific and unspecified acute bronchiolitis (RSV-specific and bronchiolitis); and (c) RSV-specific and unspecified ALRI (RSV-specific and ALRI). The study reported a mean of 36,743 yearly admissions potentially due to RSV, resulting in a mean annual cost of 87.1 million. RSV-specific codes accounted for 39.2% of cases, unspecified acute bronchiolitis for 20.1%, and other unspecified ALRI codes for the remaining 40.6%. The mean hospitalization rate per 1,000 children was 55.5 in the first year of life, 16.0 in the second, and 5.4 between 24 and 59 months. A considerable proportion of cases occurred in children under two years old (> 80.4%) and even during the first year of life (> 61.7%). Otherwise healthy children accounted for 92.9% of hospitalizations and 83.3% of costs during the period. Children born preterm accounted for 1.3% of hospitalizations and 5.7% of costs. The findings revealed that RSV still contributes to a high burden on the Spanish health care system. Children under one year of age and otherwise healthy term infants accounted for most of the substantial clinical and economic burden of RSV. Current evidence potentially underestimates the true epidemiology and burden of severe RSV infection; thus, further studies focusing on the outpatient setting are needed.
Assuntos
Bronquiolite , Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Infecções Respiratórias , Recém-Nascido , Lactente , Humanos , Criança , Estudos Retrospectivos , Estresse Financeiro , Hospitalização , Bronquiolite/epidemiologia , Hospitais PúblicosRESUMO
BACKGROUND: Influenza may trigger complications, particularly in at-risk groups, potentially leading to hospitalization or death. However, due to lack of routine testing, influenza cases are infrequently coded with influenza-specific diagnosis. Statistical models using influenza activity as an explanatory variable can be used to estimate annual hospitalizations and deaths associated with influenza. Our study aimed to estimate the clinical and economic burden of severe influenza in Spain, considering such models. METHODS: The study comprised ten epidemic seasons (2008/2009-2017/2018) and used two approaches: (i) a direct method of estimating the seasonal influenza hospitalization, based on the number of National Health Service hospitalizations with influenza-specific International Classification of Diseases (ICD) codes (ICD-9: 487-488; ICD-10: J09-J11), as primary or secondary diagnosis; (ii) an indirect method of estimating excess hospitalizations and deaths using broader groups of ICD codes in time-series models, computed for six age groups and four groups of diagnoses: pneumonia or influenza (ICD-9: 480-488, 517.1; ICD-10: J09-J18), respiratory (ICD-9: 460-519; ICD-10: J00-J99), respiratory or cardiovascular (C&R, ICD-9: 390-459, 460-519; ICD-10: I00-I99, J00-J99), and all-cause. Means, excluding the H1N1pdm09 pandemic (2009/2010), are reported in this study. RESULTS: The mean number of hospitalizations with a diagnosis of influenza per season was 13,063, corresponding to 28.1 cases per 100,000 people. The mean direct annual cost of these hospitalizations was 45.7 million, of which 65.7% was generated by patients with comorbidities. Mean annual influenza-associated C&R hospitalizations were estimated at 34,894 (min: 16,546; max: 52,861), corresponding to 75.0 cases per 100,000 (95% confidence interval [CI]: 63.3-86.3) for all ages and 335.3 (95% CI: 293.2-377.5) in patients aged ≥ 65 years. We estimate 3.8 influenza-associated excess C&R hospitalizations for each hospitalization coded with an influenza-specific diagnosis in patients aged ≥ 65 years. The mean direct annual cost of the estimated excess C&R hospitalizations was 142.9 million for all ages and 115.9 million for patients aged ≥ 65 years. Mean annual influenza-associated all-cause mortality per 100,000 people was estimated at 27.7 for all ages. CONCLUSIONS: Results suggest a relevant under-detected burden of influenza mostly in the elderly population, but not neglectable in younger people.
Assuntos
Influenza Humana , Idoso , Humanos , Estações do Ano , Influenza Humana/epidemiologia , Espanha , Medicina Estatal , Hospitalização , PandemiasRESUMO
Respiratory syncytial virus (RSV) infection is a major cause of morbidity in children. However, its disease burden remains poorly understood, particularly outside of the hospital setting. Our study aimed to estimate the burden of medically attended acute lower respiratory infection (ALRI) cases potentially related to RSV in Spanish children. Longitudinal data from September 2017 to June 2018 of 51,292 children aged < 5 years old from the National Healthcare System (NHS) of two Spanish regions were used. Three case definitions were considered: (a) RSV-specific; (b) RSV-specific and unspecified acute bronchiolitis (RSV-specific and Bronchiolitis), and; (c) RSV-specific and unspecified ALRI (RSV-specific and ALRI). A total of 3460 medically attended ALRI cases potentially due to RSV were identified, of which 257 (7.4%), 164 (4.7%), and 3039 (87.8%) coded with RSV-specific, unspecific bronchiolitis, and unspecific ALRI codes, respectively. Medically attended RSV-specific and ALRI cases per 1000 children was 134.4 in the first year of life, 119.4 in the second, and 35.3 between 2 and 5 years old. Most cases were observed in otherwise healthy children (93.1%). Mean direct healthcare cost per medically attended RSV-specific and ALRI case was 1753 in the first year of life, 896 in the second, and 683 between 2 and 5 years old. Hospitalization was the main driver of these costs, accounting for 55.6%, 38.0% and 33.4%, in each respective age group. In RSV-specific cases, mean direct healthcare cost per medically attended case was higher, mostly due to hospitalization: 3362 in the first year of life (72.9% from hospitalizations), 3252 in the second (72.1%), and 3514 between 2 and 5 years old (74.2%). These findings suggest that hospitalization data alone will underestimate the RSV infections requiring medical care, as will relying only on RSV-specific codes. RSV testing and codification must be improved and preventive solutions adopted, to protect all infants, particularly during the first year of life.
Assuntos
Infecções por Vírus Respiratório Sincicial , Vírus Sincicial Respiratório Humano , Infecções Respiratórias , Criança , Pré-Escolar , Humanos , Lactente , Estresse Financeiro , Hospitalização , Infecções por Vírus Respiratório Sincicial/epidemiologia , Infecções Respiratórias/epidemiologia , EspanhaRESUMO
Antigens of both Dirofilaria immitis and Wolbachia symbiont bacteria are implicated in the inflammatory pathology of heartworm infection. The aim of the present study was to compare the stimulatory capacity of in vitro cultures of vascular endothelial cells by the adult somatic antigens of D. immitis (DiSA) and the recombinant form of the Wolbachia surface protein (rWSP), during the first 24h of stimulation. Our results indicate a different stimulatory activity of the two antigens. Both the DiSA and rWSP stimulate the production of the enzymes responsible of the arachidonic acid metabolism, cyclooxygenase-2, 5-lipoxygenase (5-LO), and leukotriene B4. Only DiSA stimulates the production of prostaglandin E2. Related to the adhesion molecules, the DiSA stimulates the expression of intercellular adhesion molecule-1 (ICAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1), whereas rWSP stimulates ICAM-1, PECAM-1, and vascular cell adhesion molecule-1 (VCAM-1). Expression of E-cadherin and vascular endothelial growth factor also were stimulated by rWSP. Neither of the two antigens altered the basic physiological mechanisms of endothelial cells, such as cell proliferation, cell cycle, or apoptosis. The biological and pathological significance of these finding are discussed.
Assuntos
Antígenos de Helmintos/imunologia , Dirofilaria immitis/imunologia , Endotélio Vascular/citologia , Proteínas de Membrana/imunologia , Wolbachia/imunologia , Animais , Células Cultivadas , Células Endoteliais/imunologia , Células Endoteliais/metabolismo , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Humanos , Inflamação/imunologia , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/imunologia , Molécula 1 de Adesão Intercelular/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/imunologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Molécula 1 de Adesão de Célula Vascular/genética , Molécula 1 de Adesão de Célula Vascular/imunologia , Molécula 1 de Adesão de Célula Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/imunologia , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Dirofilaria immitis is the causal agent of cardiopulmonary dirofilariosis (heartworm disease). Adult worms lodge in the pulmonary arteries and right ventricle, thus vascular endothelium is exposed to high concentrations of Dirofilaria antigenic products. Heartworm disease habitually develops as a chronic foreseeable pathology. Moreover, the simultaneous death of many adult worms, naturally or induced by a filaricide treatment, can cause acute thromboembolisms and endarteritis. To better understand the effects of the massive death of D. immitis adult worms on the blood vessel endothelium, we cultured vascular endothelial cells in the presence or absence of an antigenic extract of D. immitis adult worms (DiSA). The parasite products increased the expression of enzymes and the synthesis of eicosanoids related to inflammation, such as COX-2, 5-LO, PGE(2) and LTB(4). The expression of ICAM-1 and PECAM-1 adhesion molecules and endothelial and inducible Nitric Oxide Synthases (eNOS and iNOS) was also increased in cultures treated with DiSA. Nevertheless, DiSA decreased endothelial permeability and does not alter both proliferation and apoptosis. These results suggest that the somatic extract of D. immitis adult worms stimulate inflammatory mechanisms in endothelial cells, without altering their basic physiologic processes.
Assuntos
Antígenos de Helmintos/imunologia , Dirofilaria immitis/patogenicidade , Células Endoteliais/imunologia , Endotélio Vascular/imunologia , Endotélio Vascular/fisiologia , Molécula 1 de Adesão Intercelular/metabolismo , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Animais , Permeabilidade da Membrana Celular , Células Cultivadas , Dirofilaria immitis/imunologia , Dirofilariose , Eicosanoides/metabolismo , Células Endoteliais/fisiologia , Endotélio Vascular/citologia , Humanos , Inflamação/imunologia , Óxido Nítrico Sintase/metabolismoRESUMO
Feline heartworm (Dirofilaria immitis) infection is a severe, life-threatening disease. The eicosanoids are lipid mediators derived from the metabolism of the arachidonic acid, involved in the regulation of the immune response and of inflammatory reactions. In this study, naturally infected cats showed significant higher levels of prostaglandin E(2) (PGE2), thromboxane B(2) (TXB(2)) and leukotriene B(4) (LTB4) than uninfected cats. Changes in the levels of eicosanoids during the infection were observed in experimentally infected cats. PGE2 increased significantly during the first 60 days post-infection, then progressively decreased until day 180 post-infection. At this time, PGE2 values are still significantly higher than those observed before the infection. TxB2 and LTB4 increased progressively from the beginning of infection and reached their maximum levels 180 days post-infection. In experimentally infected, ivermectin-treated cats, 15 days after treatment (45 days after infection) both PGE2 and LTB4 levels were similar to those observed in experimentally infected, untreated cats. No significant differences of PGE2 levels were found before the infection and at the end of the experiment (165 days post-treatment, 195 days post-infection). Increased levels of LTB4 were found 15 days post-treatment, afterward they progressively decreased. These data show that D. immitis infection influences the production of intravascular eicosanoids in cats. The high levels of PGE2 observed in the early phase of infection could be related to the survival of the worms, while those of TxB2 and LTB4 detected at the end of the study could mediate the inflammatory reactions and thrombi formation during the feline dirofilariosis.
Assuntos
Doenças do Gato/metabolismo , Dirofilaria immitis , Dirofilariose/metabolismo , Eicosanoides/metabolismo , Animais , Doenças do Gato/parasitologia , GatosRESUMO
The bacterial endosymbiont Wolbachia of several species of filarial nematodes plays an important role in the inflammatory pathology of filariasis. Nitric oxide (NO) production has also been implicated in the immune response during filarial infections. Here we present data indicating that a recombinant Wolbachia surface protein (rWSP) induces iNOs mRNA expression and NO production, as well as IFN-gamma and a Th1-type antibody response, in inoculated BALB/c mice. This effect is not observed when mice are inoculated with a recombinant heat shock protein from Wolbachia (GroEL).
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Dirofilaria immitis/microbiologia , Óxido Nítrico Sintase Tipo II/metabolismo , Wolbachia/imunologia , Animais , Anticorpos Antibacterianos/biossíntese , Chaperonina 60/imunologia , Dirofilaria immitis/fisiologia , Dirofilariose/imunologia , Dirofilariose/parasitologia , Inflamação/imunologia , Interferon gama/genética , Interferon gama/imunologia , Interleucina-10/genética , Interleucina-4/genética , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , RNA Mensageiro , Proteínas Recombinantes/imunologia , Simbiose , Células Th1/imunologia , Wolbachia/fisiologiaRESUMO
Dirofilaria immitis is the agent of canine heartworm disease, in which adult worms reside in the pulmonary arteries, producing first stage larvae (microfilariae) that are released into the bloodstream. The present work describes the cytokine and iNOS mRNA expression in the peripheral blood of naturally infected dogs classified as either microfilariemic or amicrofilariemic. Results show that microfilariemic dogs had higher expression of IL-4 and iNOS mRNA than amicrofilariemic dogs. Furthermore, IL-10 mRNA expression was strongly expressed in dogs with circulating microfilariae, compared to only negligible expression in amicrofilariemic dogs. Finally, mf+ status was associated with a predominance in IgG1 production against worm antigens. These results would suggest that circulating mf may stimulate, like in other filarial infections, an immune bias towards unresponsiveness in D. immitis-infected dogs, consenting long-term adult worm survival.
Assuntos
Dirofilaria immitis , Dirofilariose/imunologia , Doenças do Cão/imunologia , Interleucina-10/genética , Interleucina-4/genética , Óxido Nítrico Sintase Tipo II/genética , RNA Mensageiro/sangue , Animais , CãesRESUMO
The canine parasite Dirofilaria immitis can infect humans. Patients with pulmonary dirofilariosis develop significantly higher thromboxane B2 levels than healthy individuals living in areas where dirofilariosis is endemic and in areas where dirofilariosis is not endemic. The possible role of Wolbachia bacteria in the appearance of this eicosanoid is discussed.
Assuntos
Dirofilariose/sangue , Pneumopatias Parasitárias/sangue , Tromboxano B2/sangue , Animais , Dirofilaria immitis , Cães , Humanos , Tromboxano B2/biossínteseRESUMO
The etiologic agents of human dirofilariasis in the Old World are Dirofilaria immitis, which cause pulmonary and subcutaneous nodules, and Dirofilaria repens, which cause ocular lesions. Although reports of new cases of dirofilariasis are sporadic in other parts of the world, a considerable amount of information is generated in Europe regarding human dirofilariasis. Most cases have been detected in the Mediterranean countries, Ukraine, and Russia; however, isolated or short series of cases have been reported in the Balkan Republics and central and northern European countries. Seroepidemiologic studies have provided evidence that humans living in endemic areas present rates of infection similar to those of the autochthonous canine populations. Antibodies against endosymbiont Wolbachia bacteria have been demonstrated recently in human Dirofilaria infections. During D. immitis infections, preadult worms and third- and fourth-stage larvae are often destroyed by the host reaction, releasing a considerable amount of Wolbachia, and a Th1-type response against Wolbachia and/or filarial antigens is mounted. On the contrary, infections with D. repens, in which worms frequently remain intact, no Th1-type response has been observed. As humans are resistant hosts, the Th1-response could have a role in the resistance against parasites. The causes for the rise in the incidence of human dirofilariasis as well as the possible application of Wolbachia antigens in the serodiagnosis of human infections are discussed.
Assuntos
Anticorpos Antibacterianos/sangue , Dirofilaria/microbiologia , Dirofilariose/epidemiologia , Wolbachia/imunologia , Animais , Anticorpos Antibacterianos/análise , Dirofilaria immitis/microbiologia , Dirofilariose/sangue , Dirofilariose/parasitologia , Dirofilariose/transmissão , Europa (Continente)/epidemiologia , Humanos , Vigilância de Evento Sentinela , Estudos Soroepidemiológicos , SimbioseRESUMO
Human and animal parasitic filarial nematodes, including the agent of canine and feline heartworm disease Dirofilaria immitis, harbour intracellular bacteria of the genus Wolbachia (Rickettsiaies). It is thought that these bacteria play an important role in the pathogenesis and immune response to filarial infection. Immunoglobulin G (total IgG, IgG1, IgG2) production against and immunohistochemical staining of tissues for the Wolbachia surface protein (WSP) from dogs with natural heartworm infection were evaluated. All infected dogs had significant total anti-WSP IgG levels compared to healthy controls. Interestingly, WSP was recognized by the IgG2 subclass in both microfilariemic dogs and in dogs with no circulating microfilariae (occult infection). However, microfilariemic dogs also produced gG1 antibodies. Positive staining for WSP was observed in lungs, liver and kidneys, in particular in glomerular capillaries of naturally infected dogs who had died from heartworm disease. Our results show for the first time that Wolbachia is recognized specifically by D. immitis--infected dogs and that the bacteria is released into host tissue. Furthermore, microfilariemic status appears to effect immune responses to this endosymbiont.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Dirofilaria immitis/microbiologia , Dirofilariose/imunologia , Dirofilariose/microbiologia , Wolbachia/imunologia , Animais , Anticorpos Antibacterianos/biossíntese , Proteínas da Membrana Bacteriana Externa/metabolismo , Dirofilariose/parasitologia , Cães , Feminino , Humanos , Imunoglobulina G/biossíntese , Imuno-HistoquímicaRESUMO
This study was undertaken to investigate the enzymatic regulation of the biosynthesis of vasoconstrictor prostanoids by resting and interleukin (IL)-1(beta)stimulated human umbilical vein endothelial cells (HUVECs). Biosynthesis of eicosanoids in response to IL-1beta, exogenous labeled arachidonic acid (AA), or histamine, as well as their spontaneous release, was evaluated by means of HPLC and RIA. HUVECs exposed to IL-1beta produced prostaglandin (PG) I2 for no longer than 30 seconds after the substrate was added irrespective of the cyclooxygenase (COX) activity, whereas the time course of PGE2 and PGD2 formation was parallel to the COX activity. The ratio of PGE2 to PGD2 produced by HUVECs was similar to that obtained by purified COX-1 and COX-2. Production of PGF2alpha from exogenous AA was limited and similar in both resting and IL-1beta-treated cells. PGF2alpha was the main prostanoid released into the medium during exposure to IL-1beta, whereas when HUVECs treated with IL-1beta were stimulated with histamine or exogenous AA, PGE2 was released in a higher quantity than PGF2alpha. PGF2alpha released into the medium during treatment with IL-1beta and the biosynthesis of PGE2 and PGD2 in response to exogenous AA or histamine increased with COX-2 expression, whereas this did not occur in the case of PGI2. We observed that PGI synthase (PGIS) mRNA levels were not modified by the exposure to IL-1beta, but the enzyme was partially inactivated. When SnCl2 was added to the incubation medium, the transformation of exogenous AA-derived PGH2 into PGE2 and PGD2 was totally diverted toward PGF2alpha. Overall, these results support the conclusions that PGE2 and PGD2 (and also probably PGF2alpha) were nonenzymatically derived from PGH2 in HUVECs. The concept that a high ratio of PGH2 was released by the IL-1beta-treated HUVECs and isomerized outside the cell into PGE2 and PGD2 was supported by the biosynthesis of thromboxane B2 by COX-inactivated platelets, indicating the uptake by platelets of HUVEC-derived PGH2. The IL-1beta-induced increase in the release of PGH2 by HUVECs was suppressed by the COX-2-selective inhibitor SC-58125 and correlated with both COX-2 expression and PGIS inactivation. An approach to the mechanism of inactivation of PGIS by the exposure to IL-1beta was performed by using labeled endoperoxides as substrate. The involvement of HO. in the PGIS inactivation was supported by the fact that deferoxamine, pyrrolidinedithiocarbamate, DMSO, mannitol, and captopril antagonized the effect of IL-1beta on PGIS to different degrees. The NO synthase inhibitor NG-monomethyl-L-arginine also antagonized the PGIS inhibitory effect of IL-1beta, indicating that NO. was also involved. NO. reacts with O2-. to form peroxynitrite, which has been reported to inactivate PGIS. Homolytic fission of the O-O bond of peroxynitrite yields NO2. and HO.. The fact that 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO), which reacts with NO. to form NO2., dramatically potentiated the IL-1beta effect suggests that NO2. could be a species implicated in the inactivation of PGIS. Cooperation of HO. was supported by the fact that DMSO partially antagonized the effect of carboxy-PTIO. Although our results on the exact mechanism of the inactivation of PGIS caused by IL-1beta were not conclusive, they strongly suggest that both NO. and HO. were involved.
Assuntos
Endotélio Vascular/enzimologia , Isoenzimas/genética , Prostaglandina-Endoperóxido Sintases/genética , Prostaglandinas/biossíntese , Vasoconstrição/genética , Ácido Araquidônico/farmacologia , Células Cultivadas , Ciclo-Oxigenase 2 , Epoprostenol/biossíntese , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/fisiologia , Histamina/farmacologia , Humanos , Interleucina-1/farmacologia , Proteínas de Membrana , Prostaglandina H2 , Prostaglandinas H/genética , Especificidade por Substrato , Veias Umbilicais , Vasoconstrição/efeitos dos fármacosRESUMO
1. Cyclo-oxygenase (COX), the enzyme responsible for the conversion of arachidonic acid (AA) to prostaglandin H2 (PGH2), exists in two forms, termed COX-1 and COX-2 which are encoded by different genes. COX-1 is expressed constitutively and is known to be the site of action of aspirin and other nonsteroidal anti-inflammatory drugs. COX-2 may be induced by a series of pro-inflammatory stimuli and its role in the development of inflammation has been claimed. 2. Endothelial cells are an important physiological source of prostanoids and the selective induction of COX-2 activity has been described for finite cultures of endothelial cells, but not for permanent endothelial cell lines. 3. The HUV-EC-C line is a permanent endothelial cell line of human origin. We have determined the COX activity of these cells under basal conditions and after its exposure to two different stimuli, phorbol 12-myristate 13-acetate (PMA) and interleukin-1 beta (IL-1 beta). 4. Both PMA and IL-1 beta produced dose- and time-dependent increases of the synthesis of the COX-derived eicosanoids. These increases were maximal after the treatment with 10 nM PMA for 6 to 9 h. Under these conditions, the main eicosanoid produced by the cells was PGE2. 5. The increase of COX activity by PMA or IL-1 beta correlated with an increase of the enzyme's apparent Vmax, whilst the affinity for the substrate, measured as apparent Km, remained unaffected. 6. Treatment of the cells with PMA induced a time-dependent increase in the expression of both COX-1 and COX-2 mRNAs. Nevertheless, this increase was reflected only as an increase of the COX-2 isoenzyme at the protein level. 7. The enzymatic activity of the PMA-induced COX was measured in the presence of a panel of enzyme inhibitors, and the IC50 values obtained were compared with those obtained for the inhibition of human platelet COX activity, a COX-1 selective assay. Classical non-steroidal anti-inflammatory drugs (NSAIDs) inhibited both enzymes with varying potencies but only the three compounds previously shown to be selective COX-2 inhibitors (SC-58125, NS-398 and nimesulide) showed higher potency towards the COX of PMA-treated HUV-EC-C. 8. Overall, it appears that the stimulation of the HUV-EC-C line with PMA selectively induces the COX-2 isoenzyme. This appears to be a suitable model for the study of the physiology and pharmacology of this important isoenzyme, with a permanent endothelial cell line of human origin.
Assuntos
Endotélio/efeitos dos fármacos , Interleucina-1/farmacologia , Prostaglandina-Endoperóxido Sintases/efeitos dos fármacos , Acetato de Tetradecanoilforbol/farmacologia , Linhagem Celular , Relação Dose-Resposta a Droga , Humanos , Indometacina/farmacologia , Prostaglandina-Endoperóxido Sintases/metabolismoRESUMO
This study was focused on the characterization of the metabolism of linoleic acid by human dermal fibroblasts and the effect of interleukin-1 on the biosynthesis of octadecanoids. Dermal fibroblasts untreated and treated with recombinant IL-1beta were incubated with exogenous labeled linoleic acid. A combination of high performance liquid chromatography and gas chromatography-mass spectrometry was used as the analytic technique. We found that dermal fibroblasts convert linoleic acid mainly into 13-hydroxy-9-cis,11-trans-octadecadienoic acid (13-HODE) and 9-hydroxy-10-trans,12-cis-octadecadienoic acid (9-HODE), 13(S)-HODE and 9(R)-HODE being the predominant enantiomers. IL-1beta increased the formation of both 13-HODE and 9-HODE in a concentration-dependent manner with similar EC50 values as for prostanoid formation. This effect of IL-1beta on HODEs formation was concomitant with the expression of prostaglandin H-synthase-2. Formation of octadecanoids was inhibited in a concentration-dependent manner by acetylsalicylic acid and indomethacin. Dexamethasone, actinomycin D, and cycloheximide abolished the effect of IL-1beta on HODEs biosynthesis. Octadecanoid biosynthetic activity was associated with the microsomal fraction. Dermal fibroblasts incorporated [14C]-9-HODE and [14C]-13-HODE into phospholipids, mainly into phosphatidylcholine. IL-1beta increased significantly the esterification of 13-HODE in all glycerophospholipids, the major increase being observed in phosphatidylinositol. These results indicate that prostaglandin H-synthase-2 is the enzyme responsible for the increase in the ability to form HODEs of dermal fibroblasts stimulated with IL-1beta.
Assuntos
Interleucina-1/farmacologia , Ácidos Linoleicos Conjugados , Ácidos Linoleicos/biossíntese , Ácidos Linoleicos/metabolismo , Prostaglandina-Endoperóxido Sintases/fisiologia , Pele/metabolismo , Células Cultivadas , Feminino , Fibroblastos/metabolismo , Humanos , Ácido LinoleicoRESUMO
1. The effects of aminoguanidine on neutrophil adherence to venules and on the diameter of arterioles in the mesenteric vascular bed of the pentobarbitone-anaesthetized rat have been compared with those of the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME). 2. Administration of L-NAME (1-10 mg kg-1, i.v.) caused a dose-dependent increase in leukocyte adherence and a reduction in leukocyte rolling velocity in postcapillary venules of the rat mesentery over 1 h. 3. Likewise, aminoguanidine (10-100 mg kg-1, i.v.) dose-dependently increased leukocyte adherence and decreased leukocyte rolling velocity over 1 h. 4. Both L-NAME and aminoguanidine caused a dose-dependent reduction in mesenteric arteriolar diameter and an increase in systemic arterial blood pressure. 5. The effects of aminoguanidine (50 mg kg-1, i.v.) on leukocyte adherence, arteriolar diameter and on blood pressure were significantly reversed by pretreatment with L-arginine (300 mg kg-1, i.v.). 6. These findings indicate that, like L-NAME, aminoguanidine can acutely promote leukocyte adherence to the mesenteric venular wall and reduce arteriolar diameter. Moreover, these acute effects were reversed by L-arginine, suggesting they are mediated through inhibition of constitutive NO synthase.
Assuntos
Inibidores Enzimáticos/farmacologia , Guanidinas/farmacologia , Veias Mesentéricas/efeitos dos fármacos , Veias Mesentéricas/enzimologia , Neutrófilos/efeitos dos fármacos , Óxido Nítrico Sintase/antagonistas & inibidores , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Arteríolas/anatomia & histologia , Arteríolas/efeitos dos fármacos , Arteríolas/enzimologia , Pressão Sanguínea/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Artérias Mesentéricas/enzimologia , NG-Nitroarginina Metil Éster , Neutrófilos/citologia , Neutrófilos/fisiologia , Ratos , Ratos Wistar , Vênulas/anatomia & histologia , Vênulas/efeitos dos fármacos , Vênulas/enzimologiaRESUMO
Local intra-arterial infusion of picomole quantities of endothelin-1 induced gastric vascular leakage of radiolabelled albumin. This leakage was partially inhibited by the platelet activating factor (PAF) receptor antagonist WEB 2086 (0.5-2 mg kg-1), but was unaffected by the thromboxane synthase inhibitor OKY 1581 (5 mg kg-1) or by pretreatment with anti-neutrophil serum. These results indicate a partial role of PAF, but demonstrate that neutrophils are not involved in the gastric vascular dysfunction induced by locally administered endothelin-1.
Assuntos
Permeabilidade Capilar/efeitos dos fármacos , Endotelinas/toxicidade , Endotélio Vascular/efeitos dos fármacos , Mucosa Gástrica/irrigação sanguínea , Fator de Ativação de Plaquetas/farmacologia , Animais , Azepinas/administração & dosagem , Azepinas/farmacologia , Modelos Animais de Doenças , Endotelinas/administração & dosagem , Endotélio Vascular/lesões , Mucosa Gástrica/efeitos dos fármacos , Infusões Intra-Arteriais , Marcação por Isótopo , Masculino , Metacrilatos/administração & dosagem , Metacrilatos/farmacologia , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Fator de Ativação de Plaquetas/antagonistas & inibidores , Ratos , Ratos Wistar , Albumina Sérica/metabolismo , Úlcera Gástrica/induzido quimicamente , Tromboxano-A Sintase/antagonistas & inibidores , Triazóis/administração & dosagem , Triazóis/farmacologiaRESUMO
The effects of locally infused calcitonin gene-related peptide (CGRP) on the changes in microvascular permeability and tissue injury following close-arterial infusion of endothelin-1 (5 pmol kg-1 min-1, 10 min) in the rat gastric mucosa have been investigated. Pretreatment with rat alpha-CGRP (10-50 pmol kg-1 min-1) inhibited the leakage of radiolabelled albumin and reduced the haemorrhagic necrosis, an action inhibited by the CGRP receptor antagonist, hCGRP-(8-37) (2 nmol kg-1 min-1). By contrast, local infusion of CGRP immediately following endothelin-1 challenge had no such protective actions, and significantly exacerbated the microvascular leakage and tissue injury at the higher dose. These studies suggest that CGRP can exert both pro- and anti-inflammatory actions in the gastric microcirculation.