Seguimiento de niños con diagnóstico de asma grave antes y durante la pandemia por COVID-19 / Follow-up of children diagnosed with severe asthma before and during the COVID-19 pandemic

En la pandemia por COVID-19 se exploraron estrategias de atención para garantizar el seguimiento de niños con asma grave. Estudio prospectivo, observacional, comparativo. Se incluyeron pacientes del programa de asma grave de un hospital pediátrico de tercer nivel (n 74). Se evaluó el grado de control, exacerbaciones y hospitalizaciones durante un período presencial (PP), marzo 2019-2020, y uno virtual (PV), abril 2020-2021. En el PP, se incluyeron 74 pacientes vs. 68 (92 %) del PV. En el PP, el 68 % (46) de los pacientes presentaron exacerbaciones vs. el 46 % (31) de los pacientes en el PV (p 0,003). En el PP, se registraron 135 exacerbaciones totales vs. 79 en el PV (p 0,001); hubo una reducción del 41 %. En el PP, el 47 % (32) de los pacientes tuvieron exacerbaciones graves vs. el 32 % (22) de los pacientes en el PV (p 0,048). Hubo 91 exacerbaciones graves en el PP vs. 49 en el PV (p 0,029), reducción del 46 %. No hubo diferencias en las hospitalizaciones (PP 10, PV 6; p 0,9). La telemedicina fue efectiva para el seguimiento de pacientes con asma grave

During the COVID-19 pandemic, health care strategies were explored to ensure the follow-up of children with severe asthma. This was a prospective, observational, and comparative study. Patients in the severe asthma program of a tertiary care children's hospital were included (n: 74). The extent of control, exacerbations, and hospitalizations during an in-person period (IPP) (March 2019­2020) and an online period (OP) (April 2020­2021) was assessed. A total of 74 patients were enrolled in the IPP compared to 68 (92%) in the OP. During the IPP, 68% (46) of patients had exacerbations versus 46% (31) during the OP (p = 0.003). During the IPP, 135 total exacerbations were recorded compared to 79 during the OP (p = 0.001); this accounted for a 41% reduction. During the IPP, 47% (32) of patients had severe exacerbations versus 32% (22) during the OP (p = 0.048). A total of 91 severe exacerbations were recorded during the IPP compared to 49 during the OP (p = 0.029); the reduction was 46%. No differences were observed in terms of hospitalization (IPP: 10, OP: 6; p = 0,9). Telemedicine was effective for the follow-up of patients with severe asthma.

Follow-up of children diagnosed with severe asthma before and during the COVID-19 pandemic. / Seguimiento de niños con diagnóstico de asma grave antes y durante la pandemia por COVID-19.

During the COVID-19 pandemic, health care strategies were explored to ensure the follow-up of children with severe asthma. This was a prospective, observational, and comparative study. Patients in the severe asthma program of a tertiary care children's hospital were included (n: 74). The extent of control, exacerbations, and hospitalizations during an in-person period (IPP) (March 2019-2020) and an online period (OP) (April 2020-2021) was assessed. A total of 74 patients were enrolled in the IPP compared to 68 (92%) in the OP. During the IPP, 68% (46) of patients had exacerbations versus 46% (31) during the OP (p = 0.003). During the IPP, 135 total exacerbations were recorded compared to 79 during the OP (p = 0.001); this accounted for a 41% reduction. During the IPP, 47% (32) of patients had severe exacerbations versus 32% (22) during the OP (p = 0.048). A total of 91 severe exacerbations were recorded during the IPP compared to 49 during the OP (p = 0.029); the reduction was 46%. No differences were observed in terms of hospitalization (IPP: 10, OP: 6; p = 0,9). Telemedicine was effective for the follow-up of patients with severe asthma.

En la pandemia por COVID-19 se exploraron estrategias de atención para garantizar el seguimiento de niños con asma grave. Estudio prospectivo, observacional, comparativo. Se incluyeron pacientes del programa de asma grave de un hospital pediátrico de tercer nivel (n 74). Se evaluó el grado de control, exacerbaciones y hospitalizaciones durante un período presencial (PP), marzo 2019-2020, y uno virtual (PV), abril 2020-2021. En el PP, se incluyeron 74 pacientes vs. 68 (92 %) del PV. En el PP, el 68 % (46) de los pacientes presentaron exacerbaciones vs. el 46 % (31) de los pacientes en el PV (p 0,003). En el PP, se registraron 135 exacerbaciones totales vs. 79 en el PV (p 0,001); hubo una reducción del 41 %. En el PP, el 47 % (32) de los pacientes tuvieron exacerbaciones graves vs. el 32 % (22) de los pacientes en el PV (p 0,048). Hubo 91 exacerbaciones graves en el PP vs. 49 en el PV (p 0,029), reducción del 46 %. No hubo diferencias en las hospitalizaciones (PP 10, PV 6; p 0,9). La telemedicina fue efectiva para el seguimiento de pacientes con asma grave.

AHR signaling is induced by infection with coronaviruses.

Coronavirus infection in humans is usually associated to respiratory tract illnesses, ranging in severity from mild to life-threatening respiratory failure. The aryl hydrocarbon receptor (AHR) was recently identified as a host factor for Zika and dengue viruses; AHR antagonists boost antiviral immunity, decrease viral titers and ameliorate Zika-induced pathology in vivo. Here we report that AHR is activated by infection with different coronaviruses, potentially impacting antiviral immunity and lung epithelial cells. Indeed, the analysis of single-cell RNA-seq from lung tissue detected increased expression of AHR and AHR transcriptional targets, suggesting AHR signaling activation in SARS-CoV-2-infected epithelial cells from COVID-19 patients. Moreover, we detected an association between AHR expression and viral load in SARS-CoV-2 infected patients. Finally, we found that the pharmacological inhibition of AHR suppressed the replication in vitro of one of the causative agents of the common cold, HCoV-229E, and the causative agent of the COVID-19 pandemic, SARS-CoV-2. Taken together, these findings suggest that AHR activation is a common strategy used by coronaviruses to evade antiviral immunity and promote viral replication, which may also contribute to lung pathology. Future studies should further evaluate the potential of AHR as a target for host-directed antiviral therapy.

NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) modulation by intracellular Cl- concentration.

The impairment of the cystic fibrosis transmembrane conductance regulator (CFTR) activity induces intracellular chloride (Cl- ) accumulation. The anion Cl- , acting as a second messenger, stimulates the secretion of interleukin-1ß (IL-1ß), which starts an autocrine positive feedback loop. Here, we show that NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) are indirectly modulated by the intracellular Cl- concentration, showing maximal expression and activity at 75 mM Cl- , in the presence of the ionophores nigericin and tributyltin. The expression of PYD and CARD domain containing (PYCARD/ASC) remained constant from 0 to 125 mM Cl- . The CASP1 inhibitor VX-765 and the NLRP3 inflammasome inhibitor MCC950 completely blocked the Cl- -stimulated IL-1ß mRNA expression and partially the IL-1ß secretion. DCF fluorescence (cellular reactive oxygen species, cROS) and MitoSOX fluorescence (mitochondrial ROS, mtROS) also showed maximal ROS levels at 75 mM Cl- , a response strongly inhibited by the ROS scavenger N-acetyl-L-cysteine (NAC) or the NADPH oxidase (NOX) inhibitor GKT137831. These inhibitors also affected CASP1 and NLRP3 mRNA and protein expression. More importantly, the serum/glucocorticoid regulated kinase 1 (SGK1) inhibitor GSK650394, or its shRNAs, completely abrogated the IL-1ß mRNA response to Cl- and the IL-1ß secretion, interrupting the autocrine IL-1ß loop. The results suggest that Cl- effects are mediated by SGK1, in which under Cl- modulation stimulates the secretion of mature IL-1ß, in turn, responsible for the upregulation of ROS, CASP1, NLRP3 and IL-1ß itself, through autocrine signalling.

N-acetyl cysteine reverts the proinflammatory state induced by cigarette smoke extract in lung Calu-3 cells.

Chronic obstructive pulmonary disease (COPD) and cystic fibrosis (CF) are lethal pulmonary diseases. Cigarette consumption is the main cause for development of COPD, while CF is produced by mutations in the CFTR gene. Although these diseases have a different etiology, both share a CFTR activity impairment and proinflammatory state even under sterile conditions. The aim of this work was to study the extent of the protective effect of the antioxidant N-acetylcysteine (NAC) over the proinflammatory state (IL-6 and IL-8), oxidative stress (reactive oxygen species, ROS), and CFTR levels, caused by Cigarette Smoke Extract (CSE) in Calu-3 airway epithelial cells. CSE treatment (100 µg/ml during 24 h) decreased CFTR mRNA expression and activity, and increased the release of IL-6 and IL-8. The effect on these cytokines was inhibited by N-acetyl cysteine (NAC, 5 mM) or the NF-kB inhibitor, IKK-2 (10 µM). CSE treatment also increased cellular and mitochondrial ROS levels. The cellular ROS levels were normalized to control values by NAC treatment, although significant effects on mitochondrial ROS levels were observed only at short times (5´) and effects on CFTR levels were not observed. In addition, CSE reduced the mitochondrial NADH-cytochrome c oxidoreductase (mCx I-III) activity, an effect that was not reverted by NAC. The reduced CFTR expression and the mitochondrial damage induced by CSE could not be normalized by NAC treatment, evidencing the need for a more specific reagent. In conclusion, CSE causes a sterile proinflammatory state and mitochondrial damage in Calu-3 cells that was partially recovered by NAC treatment.

Intracellular Chloride Concentration Changes Modulate IL-1ß Expression and Secretion in Human Bronchial Epithelial Cultured Cells.

Cystic fibrosis (CF) is caused by mutations in the CFTR gene, which encodes a cAMP-regulated chloride channel. Several cellular functions are altered in CF cells. However, it is not clear how the CFTR failure induces those alterations. We have found previously several genes differentially expressed in CF cells, including c-Src, MUC1, MTND4, and CISD1 (CFTR-dependent genes). Recently, we also reported the existence of several chloride-dependent genes, among them GLRX5 and RPS27. Here, varying the intracellular chloride concentration [Cl- ]i of IB3-1 CF bronchial epithelial cells, we show that IL-1ß mRNA expression and secretion are also under Cl- modulation. The response to Cl- is biphasic, with maximal effects at 75 mM Cl- . The regulation of the IL-1ß mRNA expression involves an IL-1ß autocrine effect, since in the presence of the IL-1ß receptor antagonist IL1RN or anti-IL-1ß blocking antibody, the mRNA response to Cl- disappeared. Similar effects were obtained with the JNK inhibitor SP600125, the c-Src inhibitor PP2 and the IKK inhibitor III (BMS-345541). On the other hand, the IL-1ß secretion is still modulated by Cl- in the presence of IL-1RN, IL-1ß blocking antibody, or cycloheximide, suggesting that Cl- is affecting the IL-1ß maturation/secretion, which in turn starts an autocrine positive feedback loop. In conclusion, the Cl- anion acts as a second messenger for CFTR, modulating the IL-1ß maturation/secretion. The results also imply that, depending on its intracellular concentration, Cl- could be a pro-inflammatory mediator. J. Cell. Biochem. 118: 2131-2140, 2017. © 2016 Wiley Periodicals, Inc.

Oxidative stress-induced premature senescence dysregulates VEGF and CFH expression in retinal pigment epithelial cells: Implications for Age-related Macular Degeneration.

Oxidative stress has a critical role in the pathogenesis of Age-related Macular Degeneration (AMD), a multifactorial disease that includes age, gene variants of complement regulatory proteins and smoking as the main risk factors. Stress-induced premature cellular senescence (SIPS) is postulated to contribute to this condition. In this study, we hypothesized that oxidative damage, promoted by endogenous or exogenous sources, could elicit a senescence response in RPE cells, which would in turn dysregulate the expression of major players in AMD pathogenic mechanisms. We showed that exposure of a human RPE cell line (ARPE-19) to a cigarette smoke concentrate (CSC), not only enhanced Reactive Oxygen Species (ROS) levels, but also induced 8-Hydroxydeoxyguanosine-immunoreactive (8-OHdG) DNA lesions and phosphorylated-Histone 2AX-immunoreactive (p-H2AX) nuclear foci. CSC-nuclear damage was followed by premature senescence as shown by positive senescence associated-ß-galactosidase (SA-ß-Gal) staining, and p16(INK4a) and p21(Waf-Cip1) protein upregulation. N-acetylcysteine (NAC) treatment, a ROS scavenger, decreased senescence markers, thus supporting the role of oxidative damage in CSC-induced senescence activation. ARPE-19 senescent cultures were also established by exposure to hydrogen peroxide (H2O2), which is an endogenous stress source produced in the retina under photo-oxidation conditions. Senescent cells upregulated the proinflammatory cytokines IL-6 and IL-8, the main markers of the senescence-associated secretory phenotype (SASP). Most important, we show for the first time that senescent ARPE-19 cells upregulated vascular endothelial growth factor (VEGF) and simultaneously downregulated complement factor H (CFH) expression. Since both phenomena are involved in AMD pathogenesis, our results support the hypothesis that SIPS could be a principal player in the induction and progression of AMD. Moreover, they would also explain the striking association of this disease with cigarette smoking.

Asma por ejercicio: utilidad de marcadores en orina / Exercise induced asthma: utility of urinaria markers

Antecedentes: los metabolitos estables de la Prostaglandina D2 (PGD2) mastocitaria 9 Ó11ß Prostaglandina F2 (9 Ó11ßPGF2) y de los cis-leucotrienos (LTE4) medidos en orina reflejan la producción de estos mediadores. Objetivos: determinar el rol de los leucotrienos y de la Prostaglandina D2 a través de la relación existente entre la provocación del asma por ejercicio (AIE) y los niveles urinarios de Laucotrieno E4 (LTE4) y 9 Ó11ßPGF2. Materiales y métodos: fueron estudiados 24 niños con asma (6-14 años) y 9 niños sanos como control. En todos los asmáticos y en 5 controles se evaluó la presencia de AIE mediante prueba de carrera libre durante 7 min, alcanzando el 80 por ciento de frecuencia cardíaca máxima para la edad. Se realizaron espirometrías basales y post prueba (secuenciales) y se colectó orina inmediatamente antes y 45 minutos despues de la prueba. LTE4 y 9 Ó11ßPGF2 fue evaluada por enzimainmunoensayos específicos. Resultados: Los 5 controles normales no presentaron asma por ejercicio, de los 24 pacientes asmáticos 12 no presentaron AIE y en 12 la prueba fue posititva (VEF1s cae > 15 por ciento). Las medias de los valores basales y post ejercicio de LTE4 y 9 Ó11ßPGF2 en pg/mg creatinina se tabulan a continuación: Asma por ejercicio: 9 Ó11ßPGF2: Basal: 3,39; Post: 7,95; p=0,001; LTE4: Basal: 4,00; Post: 9,39; p=0,002. Asma sin ejercicio: 9 Ó11ßPGF2: Basal: 3,98; Post: 6,28; p=0,02; LTE4: Basal: 5,91; Post: 7,04; p=0,242. Los niveles de 9 Ó11ßPGF2 y LTE4 de los controles normales no variaron significativamente post ejercicio. Conclusión: en los pacientes con asma por ejercicio se verifica activación mastocitaria con liberación de PGD2 que se demuestra como aumento de 9 Ó11ßPGF2 urinaria, y de los leucotrienos aumento del LTE4. El aumento de LTE4 es específico para asma por ejercicio en tanto que la 9 Ó11ßPGF2 aumenta en ambos grupos

Asma por ejercicio: utilidad de marcadores en orina / Exercise induced asthma: utility of urinaria markers

Antecedentes: los metabolitos estables de la Prostaglandina D2 (PGD2) mastocitaria 9 O11ß Prostaglandina F2 (9 O11ßPGF2) y de los cis-leucotrienos (LTE4) medidos en orina reflejan la producción de estos mediadores. Objetivos: determinar el rol de los leucotrienos y de la Prostaglandina D2 a través de la relación existente entre la provocación del asma por ejercicio (AIE) y los niveles urinarios de Laucotrieno E4 (LTE4) y 9 O11ßPGF2. Materiales y métodos: fueron estudiados 24 niños con asma (6-14 años) y 9 niños sanos como control. En todos los asmáticos y en 5 controles se evaluó la presencia de AIE mediante prueba de carrera libre durante 7 min, alcanzando el 80 por ciento de frecuencia cardíaca máxima para la edad. Se realizaron espirometrías basales y post prueba (secuenciales) y se colectó orina inmediatamente antes y 45 minutos despues de la prueba. LTE4 y 9 O11ßPGF2 fue evaluada por enzimainmunoensayos específicos. Resultados: Los 5 controles normales no presentaron asma por ejercicio, de los 24 pacientes asmáticos 12 no presentaron AIE y en 12 la prueba fue posititva (VEF1s cae > 15 por ciento). Las medias de los valores basales y post ejercicio de LTE4 y 9 O11ßPGF2 en pg/mg creatinina se tabulan a continuación: Asma por ejercicio: 9 O11ßPGF2: Basal: 3,39; Post: 7,95; p=0,001; LTE4: Basal: 4,00; Post: 9,39; p=0,002. Asma sin ejercicio: 9 O11ßPGF2: Basal: 3,98; Post: 6,28; p=0,02; LTE4: Basal: 5,91; Post: 7,04; p=0,242. Los niveles de 9 O11ßPGF2 y LTE4 de los controles normales no variaron significativamente post ejercicio. Conclusión: en los pacientes con asma por ejercicio se verifica activación mastocitaria con liberación de PGD2 que se demuestra como aumento de 9 O11ßPGF2 urinaria, y de los leucotrienos aumento del LTE4. El aumento de LTE4 es específico para asma por ejercicio en tanto que la 9 O11ßPGF2 aumenta en ambos grupos (AU)