Percepciones de profesionales de salud sobre la discriminación de género: trabajo con un dispositivo audiovisual en la promoción de la salud / Perceptions of health professionals on gender discrimination: work with an audiovisual device in health promotion

INTRODUCCIÓN En las últimas décadas se produjo una transformación cualitativa en la concepción de salud pública según un concepto ampliado ligado a la idea de promoción. Los medios de comunicación resultan agentes clave para promover la salud en tanto operadores de in/visibilización de ciertas temáticas. Asumiendo la necesidad de que el sector sanitario trascienda la responsabilidad de brindar servicios médicos y que comparta su tarea con otras áreas y disciplinas, se desarrolló una herramienta de trabajo conformada por un dispositivo de análisis del discurso televisivo, orientado a mostrar cómo se construyen estereotipos de género. OBJETIVOS Contribuir al posicionamiento de los profesionales de salud como agentes activos, a fin de visibilizar y cuestionar la discriminación de género en sus prácticas dentro del espacio de la consulta y la atención sanitaria. MÉTODOS Se trabajó con un diseño exploratorio descriptivo. Se analizaron programas periodísticos televisivos, relevando representaciones sobre identidades de género que responden a patrones discriminatorios. Se confeccionó y administró un cuestionario para registrar cómo perciben la discriminación de género los profesionales de salud. Se diseñó una herramienta de trabajo que permitió detectar estereotipos de género en el discurso televisivo. RESULTADOS El trabajo realizado con profesionales de salud a partir del análisis de imágenes televisivas contribuyó a desnaturalizar estereotipos de género, que sustentan formas de desigualdad y discriminación arraigadas culturalmente, y favoreció la incorporación de una mirada crítica sobre esta problemática. DISCUSIÓN La incorporación del análisis de imágenes y del discurso audiovisual como estrategia de visibilización de la inequidad y discriminación contribuye a fortalecer la sensibilidad de género en la consulta sanitaria. Sobre la base del aporte interdisciplinario de una herramienta de trabajo innovadora, se puede mejorar la calidad de atención de los profesionales en sus prácticas y ámbitos de incidencia.

Optimización del abordaje de casos de violencia de género a partir de dispositivos audiovisuales / Optimization approach to cases of gender violence from audiovisual devices

La investigación se propuso analizar las construcciones de sentido que caracterizanlas experiencias de vida de mujeres que consultan a servicios de salud porviolencia de género, así como desarrollar una herramienta de análisis del discursoaudiovisual orientada a detectar los procedimientos a través de los cuales estelenguaje configura representaciones que favorecen la producción y reproducciónde la violencia de género. La investigación buscó transmitir los recursos empleadospara efectuar dicha detección a profesionales de centros de salud.ObjetivosContribuir al posicionamiento de los servicios de salud como espacios de relevanciaen el abordaje de la violencia de género.MétodosSe trabajó con un diseño exploratorio-descriptivo. Se analizaron los estudiossobre rutas críticas realizados en Argentina, relevando las representaciones yconstrucciones de sentido que caracterizan a estas historias de vida. Se realizaronentrevistas semi-estructuradas a profesionales de servicios de salud que abordancasos de violencia de género. Se desarrolló un dispositivo audiovisual que permitióidentificar imágenes y estereotipos de género en el discurso fílmico.ResultadosEl análisis de los datos permitió identificar correspondencias directas entrelos estudios sobre rutas críticas realizados en la Argentina y estudios similaresrealizados en América Central, del Sur y Latina. El trabajo llevado a cabo a partirdel dispositivo audiovisual con profesionales de centros de salud contribuyó adesnaturalizar el lugar de subordinación que ocupan las mujeres con relacióna los hombres y a que los profesionales pudieran reflexionar sobre sus propiosestereotipos vinculados con la violencia de género.

Effect of delayed scanning on imaging and on the diagnostic accuracy of vertical root fractures in two photostimulable phosphor plates digital systems.

AIM: To evaluate the influence of delayed scanning on images obtained with two PSPs digital systems and on the diagnostic accuracy of vertical root fracture (VRF) by means of objective and subjective analyses. METHODOLOGY: Forty single-rooted human teeth were divided into two groups, one without VRFs and another with VRFs induced by a universal testing machine. Two digital systems (VistaScan(®) and Express(®) ) were used to radiograph all teeth, and the resulting plates were scanned at four time-points: T0-immediately, T1-30 min, T2-2 h and T3-4 h after exposure. An aluminium (Al) wedge was used to evaluate the change in mean grey values as each scan was delayed. Three observers screened all images for VRFs, and one-fourth of the sample was revaluated after thirty days. Areas under the receiver operating characteristic (ROC) curve, sensitivity, specificity and accuracy values were compared by anova. RESULTS: Intra- and interobserver agreement ranged from moderate to substantial and fair to moderate, respectively. There was no significant difference amongst scan delays with regard to sensitivity, specificity and accuracy; however, there were significant differences in the area under the ROC curve, with the 4-h delayed scan being associated with lower values compared to the others (P = 0.019). As for objective analysis, there was a significant difference amongst all different scanning time-points for the two systems (P = 0.001), except between the 30-min and 2-h delayed scans in the VistaScan(®) system. CONCLUSION: Whilst delayed scanning caused changes to the density of images acquired with the systems studied, it did not seem to interfere with VRF diagnosis except when scanning was delayed for 4 h, which should therefore be avoided.

Efficacy of erdosteine 900 versus 600 mg/day in reducing oxidative stress in patients with COPD exacerbations: Results of a double blind, placebo-controlled trial.

BACKGROUND: Acute exacerbations of chronic obstructive pulmonary disease (AECOPD) are associated with increased airway and systemic inflammation. There is evidence that erdosteine accelerates recovery from AECOPD by reducing airway inflammation. AIM: To investigate the dose-dependent antioxidant/anti-inflammatory activity of erdosteine in COPD patients. METHODS: In this single-centre, double blind, double dummy study, patients with mild-to-moderate COPD (GOLD stage II-III), were randomised to receive either placebo or two dosages of oral erdosteine (300 mg tid or 300 mg bid + 1 capsule of indistinguishable placebo) for 28 days in addition to their standard treatment. Primary variables were plasma reactive oxygen species (ROS) and 8-isoprostane levels, while secondary variable was lung function (FEV1; FEV1/FVC; FEV1 short-term reversibility), all assessed in baseline; every two weeks during the study, and one week after the end of the study. RESULTS: Baseline demographic characteristics, plasma ROS and 8-isoprostane levels and lung function were not significantly different in the 24 eligible patients (14 males, aged 38-75 years). At 2 weeks, there was a dose-dependent decrease in ROS in the erdosteine groups. By week 4 there were significant differences in ROS levels compared to baseline between patients receiving 900 mg/day (p < 0.003) and those receiving 600 mg/day (p < 0.04). This effect continued in the follow-up week (p < 0.021). Erdosteine also lowered 8-isoprostane plasma levels after 4 weeks (p < 0.01), and this effect lasted over the post-treatment week. Moreover, % FEV1 reversibility after salbutamol 400 mcg obtained after a 4 -week treatment of erdosteine 900 mg/day was significantly higher than that obtained after 600 mg/day (p < 0.01). Erdosteine was well tolerated and no treatment-related adverse event was reported. CONCLUSIONS: Results confirm the antioxidant dose- and time-dependent activity of erdosteine, and support the utility of including erdosteine it in the therapeutic strategy for the prevention and treatment of oxidative stress-induced inflammation, which frequently leads to AECOPD occurrence.

Influence of CBCT enhancement filters on diagnosis of vertical root fractures: a simulation study in endodontically treated teeth with and without intracanal posts.

OBJECTIVES: To evaluate the influence of CBCT enhancement filters on the diagnosis of vertical root fractures (VRFs) in teeth with and without metal posts. METHODS: The crowns of 40 uniradicular human teeth were removed and all roots were prepared. 20 teeth were randomly selected, and VRFs were induced using a universal testing machine. The i-CAT (Imaging Sciences International, Hatfield, PA) CBCT was used to scan teeth with and without intracanal metal posts using the following parameters: 0.2 voxel size, 8 × 8-cm scan size and acquisition time of 26.9 s. Images were evaluated by three observers with and without the use of the following filters: S9, smooth, smooth 3 × 3, sharpen, sharpen-mild and sharpen 3 × 3. RESULTS: Intra- and interobserver agreement ranged from poor to moderate. Images with and without CBCT filters did not show significant differences regarding the area under the receiver operating characteristic curve, as well as sensitivity (p > 0.05). As for accuracy, the sharpen-mild filter was superior to the sharpen (p = 0.03), but these filters did not differ from all others. For specificity, S9, smooth and original images were superior to sharpen (p < 0.01). Results for teeth without posts differed from those for teeth with metal posts in all cases (p < 0.05). CONCLUSIONS: The use of enhancement filters in CBCT images has no influence on the diagnosis of VRFs in teeth with metal posts, and their use is not justified.

Protective effect of quercetin against in vitro erythrocyte rheology alterations produced by arsenic.

Humans are exposed to heavy metals such as arsenic (As), through contaminated food and drinking water. The effect of As on RBC membrane is one of the most important biological effects. In a previous work, we have studied the AsVin vitro effect on erythrocytes biophysical properties discovering alterations regarding aggregability deformability, cell morphology, membrane fluidity and osmotic response. We have also observed that the presence of the metal produces an oxidative stress in RBCs that might be the origin of rheological impairment. In the present work we analyzed RBCs rheological properties associated with membrane fluidity and lipid peroxidation in presence of As and quercetin (Qc). From our results we can conclude that RBCs treatment with Qc is efficient to prevent the impairment of the mechanical properties of the cell membrane produced by the As, through oxygen reactive agents in the membrane structure, mainly on the lipids. This protective effect is observed in the preservation of the erythrocytes rheological properties and consequently in the maintenance of an appropriate blood flow, specially in the small vessels in the peripheral circulation.

Influence of maxillomandibular positioning in cone beam computed tomography for implant planning.

The aim of this study was to evaluate the influence of changes in maxillomandibular positioning during cone beam computed tomography (CBCT) imaging on the planning of dental implants. Ten skulls were marked bilaterally with metal spheres in four regions: incisors, canine, premolars, and molars. CBCT scans were obtained in seven positions: standard position (SP), displacements of 10° and 20° above and below the SP, and lateral displacements of 10° and 20° from the SP. Subsequently, bilateral measurements of the height and width of the maxilla and mandible were performed on all images. The results showed that the position with a displacement of 20° above the SP presented the greatest differences in the measurements of bone height and width. In the bilateral comparisons, the maxillary bone width showed the greatest differences, especially for the regions of the premolars and molars. It is concluded that alterations of positioning during the acquisition of CBCT images can lead to alterations in the measurements of bone height and width, which may result in errors in implant planning and cause damage to anatomical structures.

The expression of melanopsin and clock genes in Xenopus laevis melanophores and their modulation by melatonin

Vertebrates have a central clock and also several peripheral clocks. Light responses might result from the integration of light signals by these clocks. The dermal melanophores of Xenopus laevis have a photoreceptor molecule denominated melanopsin (OPN4x). The mechanisms of the circadian clock involve positive and negative feedback. We hypothesize that these dermal melanophores also present peripheral clock characteristics. Using quantitative PCR, we analyzed the pattern of temporal expression of Opn4x and the clock genes Per1, Per2, Bmal1, and Clock in these cells, subjected to a 14-h light:10-h dark (14L:10D) regime or constant darkness (DD). Also, in view of the physiological role of melatonin in the dermal melanophores of X. laevis, we determined whether melatonin modulates the expression of these clock genes. These genes show a time-dependent expression pattern when these cells are exposed to 14L:10D, which differs from the pattern observed under DD. Cells kept in DD for 5 days exhibited overall increased mRNA expression for Opn4x and Clock, and a lower expression for Per1, Per2, and Bmal1. When the cells were kept in DD for 5 days and treated with melatonin for 1 h, 24 h before extraction, the mRNA levels tended to decrease for Opn4x and Clock, did not change for Bmal1, and increased for Per1 and Per2 at different Zeitgeber times (ZT). Although these data are limited to one-day data collection, and therefore preliminary, we suggest that the dermal melanophores of X. laevis might have some characteristics of a peripheral clock, and that melatonin modulates, to a certain extent, melanopsin and clock gene expression.

The expression of melanopsin and clock genes in Xenopus laevis melanophores and their modulation by melatonin.

Vertebrates have a central clock and also several peripheral clocks. Light responses might result from the integration of light signals by these clocks. The dermal melanophores of Xenopus laevis have a photoreceptor molecule denominated melanopsin (OPN4x). The mechanisms of the circadian clock involve positive and negative feedback. We hypothesize that these dermal melanophores also present peripheral clock characteristics. Using quantitative PCR, we analyzed the pattern of temporal expression of Opn4x and the clock genes Per1, Per2, Bmal1, and Clock in these cells, subjected to a 14-h light:10-h dark (14L:10D) regime or constant darkness (DD). Also, in view of the physiological role of melatonin in the dermal melanophores of X. laevis, we determined whether melatonin modulates the expression of these clock genes. These genes show a time-dependent expression pattern when these cells are exposed to 14L:10D, which differs from the pattern observed under DD. Cells kept in DD for 5 days exhibited overall increased mRNA expression for Opn4x and Clock, and a lower expression for Per1, Per2, and Bmal1. When the cells were kept in DD for 5 days and treated with melatonin for 1 h, 24 h before extraction, the mRNA levels tended to decrease for Opn4x and Clock, did not change for Bmal1, and increased for Per1 and Per2 at different Zeitgeber times (ZT). Although these data are limited to one-day data collection, and therefore preliminary, we suggest that the dermal melanophores of X. laevis might have some characteristics of a peripheral clock, and that melatonin modulates, to a certain extent, melanopsin and clock gene expression.

Reference urinary LTE4 levels in normal individuals: a pilot study.

UNLABELLED: The definition of reference normal values for urinary LTE4 still represents an open question. AIM: to assess the influence of gender and age on urinary LTE4 levels in normal individuals. METHODS: after their informed consent, urinary LTE4 was measured in 124 well matched, non smoker, non atopic subjects (mean age 49.5 y +/- 20.1 sd; range 4-85 y, 57 m;) without any clinically evident disease and not taking any drug for several months. In all subjects, urine were collected in the morning, and processed by an immunoenzimatic method (Cayman Chem, Mi, USA) via the Triturus system (Grifols, Spain). STATISTICS: t test, anova, linear regression, assuming p < 0.05. RESULTS: mean urinary LTE4 were 57.3 pg/ml in males (mean age 51.2 y +/- 21.3 sd) and 57.0 pg/ml in females (mean age 48.1 y + 19.1 sd), p = ns. Linear regression showed no relationship between urinary LTE4 levels and subjects' age in the whole sample of subjects. When subjects were divided according to 4 different classes of age (0-14; 15-40; 41-60; > 60), anova proved that mean urinary LTE4 levels were significantly different in the different classes of age, being higher in younger subjects (67.1 pg/ml +/- 33.4 sd; 69.8 pg/ml +/- 27.5 sd; 57.1 pg/ml +/- 25.4 sd, and 45.1 pg/ml +/- 24.9, respectively) (anova p < .002; Welch test p < .005). CONCLUSIONS: 1) gender does not affect urinary LTE4 levels in normals; 2) mean urinary LTE4 concentrations tend to a slight, but significant, decrease with the increase of the subjects' age, and this is clear in those over-60; 3) reference values for younger and older normal subjects (such as, under- and over-60 years) should be assumed accordingly.

Erdosteine affects eicosanoid production in COPD.

UNLABELLED: Oxidant-antioxidant imbalance and lipid peroxidation are known to activate the 5-LO pathway with increased expression of inflammatory eicosanoids. Erdosteine has recently shown important anti-oxidant properties, including the ability to reduce 8-isoprostane in COPD patients. AIM: To assess the effects of erdosteine (E) on eicosanoids, and to compare the time-course of effect with that of E anti-oxidant activity. METHODS: 12 moderate COPD patients (9 males, 60 - 78 y) randomly received E 300 mg b.i.d. or placebo (P) for 10 days in a double-blind, controlled design. Blood ROS (Fort/Units); serum LTB4 and urine LTE4 (pg/ml) were measured at baseline and after 1, 3, 5 and 10 days of treatment. Analysis of covariance (ANCOVA) was performed. RESULTS: In COPD patients, both LTB4 and LTE4 dropped significantly during the 10-day treatment with E: s-LTB4 from 136.0 ± 35.4 SD to 54.5 ± 31.2 SD; u-LTE4 from 267.0 ± 91.5 SD to 84.0 ± 64.7 SD, p < 0.001 vs. p from Days 5 and 3, respectively. Moreover, a significant decrease of blood ROS was confirmed in patients using E. FEV1 values slightly increased during erdosteine treatment, whereas a trend to decrease was observed in the placebo group, with a significant difference in favor of erdosteine after 10 days of treatment (p = 0.0088). CONCLUSIONS: 1) The scavenging and anti-inflammatory effects of Erdosteine were both confirmed; 2) erdosteine proved to affect eicosanoids significantly; 3) this novel effect underlines the important anti-inflammatory potentialities of the drug in COPD; 4) further investigation is needed in order to assess the capability of Erdosteine in controlling ongoing inflammation in chronic respiratory diseases.

The prevalence of nasal polyps and the corresponding urinary LTE4 levels in severe compared to mild and moderate asthma.

BACKGROUND: Several comorbid conditions may contribute to worsening asthma symptoms, including nasal polyps (NPs). Cysteinyl leukotrienes (Cys-LTs) play a crucial role in asthma pathophysiology, and specific receptors for CysLTs are reported as upregulated in nasal polyp tissues. The aim of the present study was to assess the prevalence of nasal polyps in severe vs. mild and moderate asthma, and to compare the corresponding levels of urinary Leukotriene E4 (LTE4). MATERIALS AND METHODS: A cohort of 386 asthma patients were studied: n=166 with mild, n=146 with moderate and n=74 severe asthma. All patients performed a nasal endoscopy and urine were collected in the morning for the quantitative LTE4 immunoenzimatic assay (Cayman Chemical, MI, USA). Intolerance to ASA was also assessed by means of a nasal provocation test with L-ASA. RESULTS: The prevalence of NPs was the following: 8 cases (4.8%) in mild; 14 (9.6%) in moderate, and 33 (44.6%) in severe asthma. Mean urinary LTE4 levels were increasing according to the disease severity. ASA-intolerance was assessed in 1 patient in mild asthma (0.6%), 14 in moderate asthma (9.6%) and 28 in severe asthma (37.8%). CONCLUSIONS: Nasal polyps represent a comorbid which is highly frequent in severe asthma. Both their prevalence and the corresponding mean LTE4 levels in urine proved in strict, direct relationship with asthma severity. In severe asthma, nasal polyps represent a condition which is associated with the highest excretion of urinary LTE4 and ASA intolerance.

Changes in blood ROS, e-NO, and some pro-inflammatory mediators in bronchial secretions following erdosteine or placebo: a controlled study in current smokers with mild COPD.

UNLABELLED: Anti-oxidant interventions consist in reduction of direct oxidant damage by removing oxidant agents and/or by supplementing reducing agents with anti-oxidant effects. AIM: Aim of the present study was to investigate the anti-oxidant effects of erdosteine, a recent drug currently used in chronic obstructive pulmonary disease (COPD) for its rheological activity. At present, no data are available on current smokers with COPD to our knowledge. METHODS: Two groups of 10 persons matched for sex; age (65.0 yr+/-8.4 S.D. and 65.3 yr+/-6.5 S.D.); basal FEV1 (88.7% pred +/-6.8 S.D. and 85.2% pred +/-5.8 S.D.); and cigarette consumption (25.4 pack/yr+/-3.5 S.D. and 28.1 pack/yr+/-2.3 S.D.) entered a controlled, double blind, parallel groups study. They were randomized to receive erdosteine 600 mg daily or placebo for 10 days. IL-6; IL-8; TNFalpha were measured in bronchial secretions in bsln, after 4, 7, and 10 days of Erdosteine or placebo; e-NO and both ROS and 8-Isoprostane in blood were also measured at the same experimental times. STATISTICS: ANOVA: a t-test with Bonferroni correction; p<0.05 was accepted. RESULTS: Blood ROS and IL-8 in bronchial secretions dropped significantly following erdosteine starting from day 4 (both p<0.01), while 8-isoprostane drop was significant only after day 10 (p<0.02), and the e-NO decrease proved evident but not significant. No significant changes were observed in the placebo group. CONCLUSIONS: Erdosteine affects substantially some pro-inflammatory cytokines specifically involved in oxidative stress in current smokers with mild COPD. Effects appeared differently time-dependent. Further long-term studies are needed to confirm these pilot data and to assess their long-term clinical relevance.

Urinary LTE4 is higher after nasal provocation test with L-ASA in bronchial than in only nasal responders.

UNLABELLED: Aspirin induced asthma (AIA) is a syndrome characterised by intolerance to acetylsalycilic acid (ASA), nasal polyps and bronchial asthma, being the metabolic shift of arachidonic acid toward the lipoxygenase pathway and hyper-production of cysteinyl-leukotrienes (cys-LTs) the current pathogenetic hypothesis. The research for both sensitive indicators and safe diagnostic tests is still attracting. Aim of the study was to compare the levels of urinary LTE4 in baseline and after Nasal Provocation Test (NPT) with L-ASA from subjects affected by aspirinin-Intolerance and characterized by only a nasal response to ASA to those from subjects with both a nasal and a bronchial response to the same challenge. METHODS: After their written consent, 74 subjects with mill to moderate AIA (16 male, mean age 45.3 years +/- 12.3 sd, mean basal FEV1 = 78.1% pred. +/- 6.2.4sd, FEV1 reversibility = 14.3% bsln +/- 2.1 ds after salbutamol 200 mcg) performed a NPT with L-ASA (total maximal dose 25 mg). Spirometry (FEV1), acoustic rinometry (nasal volume--VOL; nasal Resistance--Req; AR; TM Hood Lab., USA), and urinary LTE4 (Cayman Chemical, MI, USA, via Triturus System, Grifols, Spain) were checked in all subjects in basal conditions and 90' after NPT. STATISTICS: t test between means +/- sd, assuming p < 0.05, and linear regression between all variables considered. RESULTS: In 69 ASA-intolerant-asthmatics, mean FEV1 did not change significantly following NPT (78.7% pred. +/- 5.1 sd in baseline; 78.5% pred. +/- 4.1 sd after NPT, p = ns) even though in the presence of a significant decrease of VOL. (12.6 cm3 +/- 4.1 sd in baseline; 6.2 cm3 +/- 4.6 sd after NPT, p = 0.003); of a substantial increase in Req (0.9 cm H2O/l/min +/- 0.1 ds in baseline; 2.4 cmH2O/l/min +/- 0.2 after NPT, p = 0.04), and of urinary LTE4 excretion (333.0 pg/mg +/- 161.7 in bsln; 558.0 pg/mg +/- 171.690' after NPT with L-SA, p = 0.02). In only 5 subjects, the nasal response occurred concomitantly to a significant bronco-constriction after the NPT: mean FEV, changed from 77.9% pred. +/- 3.9 in bsln to 46.6% pred. +/- 4.3 after NPT (p < 0.001); mean VOL from 13.9 cm3 +/- 4.7 sd to 5.6 cm3 +/- 2.8 sd (p < 0.001); mean Req from 1.1 cmH2O/l/min +/- 0.2 in bsln to 2.5 cmH2O/l/min +/- 0.4 after NPT (p = 0.02) in these subjects. In ASA-intolerant bronchial responders, the severity of respiratory reactions proved related to the extent of urinary LTE4 response, which on the other hand, proved significantly higher than that observed in ASA-intolerant subjects with only nasal response and in ASA-tolerant subjects (LTE4 from 333.0 pg/mg +/- 161.7 in baseline up to 558.0 pg/mg +/- 171.6 90 min. following the NPT with L-ASA the nasal-responders, p = 0.04, but from 412.0 pg/mg +/- 102.8 in baseline up to 978.0 pg/mg +/- 108.7 after NPT in bronchial responders, p < 0.001 from baseline). CONCLUSIONS: Nasal challenge with ASA affects significantly both nasal VOL and Req, and LTE4 excretion in all ASA-intolerant subjects. During the nasal challenge, severity of respiratory reactions proves associated with the highest basal LTE4 synthesis. This feature reflects a spectrum of respiratory tract reactions where cysteinil-LTs can play a specific diagnostic role.

Aspirin induced asthma (AIA) with nasal polyps has the highest basal LTE4 excretion: a study vs AIA without polyps, mild topic asthma, and normal controls.

UNLABELLED: Cysteinil Leukotrienes (LTs) are products of the arachidonic acid cascade which are synthetised by 5-lipoxigenase in inflammatory cells, particularly in eosinophils. Urinary leukotriene E4 concentration (LTE4), that reflects the whole body production of cysteinil-leukotrienes, is particularly increased in patients with aspirin-intolerant asthma (AIA). Aim of the present study was to assess basal urinary LTE4 levels from AIA patients with nasal polyps to those from AIA patients with only rhinitis (without polyps), and those from mild atopic asthmatics and normal controls. SUBJECTS & METHODS: 34 normal subjects (N; 19 - 57y, FEV1 = 102.1% pred. +/- 8.2 sd; negative MCh challenge; negative prick test); 39 mild-persistent atopic asthmatics (A; 18-66y, FEV1 = 92.1 %pred. +/- 14.6 sd; PD20 FEV1 = 380.7mcg +/- 481.2 sd); 24 subjects with AIA with rhinitis (AIA/R; 18 - 56y, FEV1 = 71.6%pred +/- 15.5 sd; reversibility = 15.1% bsln +/- 2.1 sd after salbutamol 200mg), and 10 subjects with AIA and nasal polyposis (AIA/NP; 22-49 y; FEV1 = 70.6%pred. +/- 7.1 sd; reversibility = 13.2% bsln +/- 1.6 sd after salbutamol 200 microg) were studied. After their informed consent, urine were collected in the morning for the LTE4 quantitative immunoenzimatic assay (pg/mg creatinine; Cayman Chemical, Ann Arbour, Mi, USA). STATISTICS: Wilcoxon signed rank test was used, and p<0.05 accepted as the lowest level of statistical significance. RESULTS: AIA/NP subjects had the highest levels of urinary LTE4 (432.3 pg/mg +/- 88.1 sd) compared to AIA/R (330.7 pg/mg +/- 72.3s, p < 0.01), to A (129.1 pg/mg +/- 74.8sd, p < 0.001), and to N controls (66.5 pg/mg +/- 20.6 sd, p < 0.001). Moreover, urinary LTE4 levels measured in AIA/R subjects proved significantly higher than those measured in A (p < 0.001) and in N controls (p<0.001), while LTE4 levels in A proved significantly higher than those in N controls (p<0.001). Furthermore, basal LTE4 levels seem inversely related to those of basal FEV1 (102.1 % pred. +/- 8.2sd in N, 92.1 % pred +/- 14.6 sd in A, 71.6 % pred. +/- 15.5 sd in AIA/R, 70.6 % pred +/- 7.1 sd in AIA/P, respectively). Respiratory function in the two sub-groups of AIA patients proved reduced than in atopic asthmatics (p<0.001) and in normal controls (p < 0.001), even though the difference between these two subgroups of subjects did not reach the statistical significance. CONCLUSIONS: Cys-LTs confirm their relevant pathogenetic role in AIA, but also in early stages of atopic asthma. Urinary LTE4 exexcretion proves directly proportional to the extent of nasal structural changes occurring in ASA-intolerant asthmatics, being subjects with nasal polyps those with the highest LTE4 values, immediately followed by those with hypertrophic rhinitis. Routinary measurements of urinary LTE4 should be regarded as a sensitive indicator in monitoring the clinical evolution of nasal involvement in AIA.

Biological activity and binding of estradiol to SK-Mel 23 human melanoma cells.

Patients expressing estradiol receptors in melanoma cells have been reported to have a better prognosis. We therefore decided to investigate the in vitro effects of beta-estradiol and tamoxifen on the growth and tyrosinase activity of SK-Mel 23 human melanoma cells. Twenty-four-hour treatment with 0.4 nM beta-estradiol inhibited cell proliferation in 30% (0.70 +/- 0.03 x 10(5) cells) and increased tyrosinase activity in 50% (7130.5 +/- 376.5 cpm/10(5) cells), as compared to untreated cells (1.0 +/- 0.05 x 10(5) cells and 4769 +/- 25.5 cpm/10(5) cells, respectively). Both responses were completely (100%) blocked by 1 microM tamoxifen. Higher concentrations (up to 1.6 nM) or longer treatments (up to 72 h) did not result in a larger effect of the hormone on proliferation or tyrosinase activity. Competition binding assays demonstrated the presence of binding sites to [2,4,6,7-3H]-beta-estradiol, and that the tritiated analogue was displaced by the unlabeled hormone (1 nM to 100 microM, Kd = 0.14 microM, maximal displacement of 93%) or by 10 microM tamoxifen (displacement of 60%). Beta-estradiol also increased the phosphorylated state of two proteins of 16 and 46 kDa, after 4-h treatment, as determined by Western blot. The absorbance of each band was 1.9- and 4-fold the controls, respectively, as determined with Image-Pro Plus software. Shorter incubation periods with beta-estradiol did not enhance phosphorylation; after 6-h treatment with the hormone, the two proteins returned to the control phosphorylation levels. The growth inhibition promoted by estradiol may explain the better prognosis of melanoma-bearing women as compared to men, and open new perspectives for drug therapy.

Biological activity and binding of estradiol to SK-Mel 23 human melanoma cells

Patients expressing estradiol receptors in melanoma cells have been reported to have a better prognosis. We therefore decided to investigate the in vitro effects of á-estradiol and tamoxifen on the growth and tyrosinase activity of SK-Mel 23 human melanoma cells. Twenty-four-hour treatment with 0.4 nM á-estradiol inhibited cell proliferation in 30 percent (0.70 ñ 0.03 x 10(5) cells) and increased tyrosinase activity in 50 percent (7130.5 ñ 376.5 cpm/10(5) cells), as compared to untreated cells (1.0 ñ 0.05 x 10(5) cells and 4769 ñ 25.5 cpm/10(5) cells, respectively). Both responses were completely (100 percent) blocked by 1 æM tamoxifen. Higher concentrations (up to 1.6 nM) or longer treatments (up to 72 h) did not result in a larger effect of the hormone on proliferation or tyrosinase activity. Competition binding assays demonstrated the presence of binding sites to [2,4,6,7- H]-á-estradiol, and that the tritiated analogue was displaced by the unlabeled hormone (1 nM to 100 æM, Kd = 0.14 æM, maximal displacement of 93 percent) or by 10 æM tamoxifen (displacement of 60 percent). á-estradiol also increased the phosphorylated state of two proteins of 16 and 46 kDa, after 4-h treatment, as determined by Western blot. The absorbance of each band was 1.9- and 4-fold the controls, respectively, as determined with Image-Pro Plus software. Shorter incubation periods with á-estradiol did not enhance phosporylation; after 6-h treatment with the hormone, the two proteins returned to the control phosphorylation levels. The growth inhibition promoted by estradiol may explain the better prognosis of melanoma-bearing women as compared to men, and open new perspectives for drug therapy.

The role of calcium, calcium-activated K+ channels, and tyrosine/kinase in psoralen-evoked responses in human melanoma cells.

8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 micro M). The intracellular Ca2+ chelator BAPTA/AM (1 micro M) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 micro M protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92% increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.

The role of calcium, calcium-activated K+ channels, and tyrosine/kinase in psoralen-evoked responses in human melanoma cells

8-Methoxy psoralen (8-MOP) exerts a short-term (24 h) mitogenic action, and a long-term (48-72 h) anti-proliferative and melanogenic action on two human melanoma cell lines, SK-Mel 28 and C32TG. An increase of intracellular calcium concentration was observed by spectrofluorometry immediately after the addition of 0.1 mM 8-MOP to both cell lines, previously incubated with calcium probe fluo-3 AM (5 µM). The intracellular Ca2+ chelator BAPTA/AM (1 µM) blocked both early (mitogenic) and late (anti-proliferative and melanogenic) 8-MOP effects on both cell lines, thus revealing the importance of the calcium signal in both short- and long-term 8-MOP-evoked responses. Long-term biological assays with 5 and 10 mM tetraethylammonium chloride (TEA, an inhibitor of Ca2+-dependent K+ channels) did not affect the responses to psoralen; however, in 24-h assays 10 mM TEA blocked the proliferative peak, indicating a modulation of Ca2+-dependent K+ channels by 8-MOP. No alteration of cAMP basal levels or forskolin-stimulated cAMP levels was promoted by 8-MOP in SK-Mel 28 cells, as determined by radioimmunoassay. However, in C32TG cells forskolin-stimulated cAMP levels were further increased in the presence of 8-MOP. In addition, assays with 1 µM protein kinase C and calcium/calmodulin-dependent kinase inhibitors, Ro 31-8220 and KN-93, respectively, excluded the participation of these kinases in the responses evoked by 8-MOP. Western blot with antibodies anti-phosphotyrosine indicated a 92 percent increase of the phosphorylated state of a 43-kDa band, suggesting that the phosphorylation of this protein is a component of the cascade that leads to the increase of tyrosinase activity.

Montelukast 10 mg improves nasal function and nasal response to aspirin in ASA-sensitive asthmatics: a controlled study vs placebo.

UNLABELLED: Aspirin-induced asthma (AIA) is a clinical syndrome characterized by acute airway reaction to aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDS). The most recent etiological hypothesises is that an overexpression of the enzyme LTC(4) synthase occurs in AIA, with the consequent production of sulfidopeptide leukotrienes (LTs). AIM: Aim of the present study was to assess the effect of Montelukast, a selective cys-LT receptor antagonist, on nasal function, nasal reactivity to ASA and blood markers of eosinophilic inflammation in mild-to-moderate AIA. MATERIAL AND METHOD: Thirty-six nonsmoker subjects with AIA (17 males, 22-52 years) performed a nasal provocation test (NPT) with lysine-aspirin (L-ASA) in baseline and after a 4-week Montelukast 10 mg or placebo treatment. Nasal function was assessed by the acoustic rhinomanometry, and they also performed a lung function test (forced expiratory volume in 1 s), and a blood sample for the eosinophil count and the eosinophil cationic protein (ECP) plasma measurements. After both treatments, all subjects repeated the NPT, the lung function, and the ECP and the eosinophil blood count. STATISTICAL ANALYSIS: t-Test was used to compare mean values +/- SD between groups, and P < 0.05 was assumed as the level for statistical significance. RESULTS: Airway patency was never affected by the NPT with L-ASA. In baseline, NPT with L-ASA precipitated a nasal reaction in all subjects, with a substantial increase in nasal resistance (calculated resistance [REQ]; from 0.89 +/- 0.18 to 2.2 +/- 0.17 cmH(2)O/l/min in group M, P < 0.001; and from 0.91 +/- 0.48 to 2.3 +/- 0.21 cmH(2)O/l/min in group P, P < 0.001); and a significant reduction in total nasal volume in at least one nostril (volume [VOL]; from 11.1 +/- 3.2 to 8.1 +/- 4.1 cm(3) in the group M, P < 0.001, and from 12.3 +/- 4.1 to 7.9 +/- 4.5 cm(3) in the group P, P < 0.001). The nasal reaction to L-ASA remained unchanged following placebo, but it was completely minimized following a 4-week treatment with Montelukast. Also nasal function, the nasal symptom score, and the markers of eosinophilic inflammation proved significantly affected and improved by the active drug only. CONCLUSIONS: Montelukast 10 mg daily for 4 weeks, but not placebo, improves nasal function and nasal response to Aspirin substantially in ASA-sensitive asthmatics.