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OBJETIVO: Determinar la carga económica anual del asma, desde una perspectiva institucional y con base en la clasificación recomendada por GINA, en una cohorte retrospectiva de adultos atendidos en el Instituto Nacional de Enfermedades Respiratorias (INER) de México. MÉTODOS: Estudio observacional, longitudinal y retrospectivo, llevado a cabo a partir de la información recabada de 247 pacientes femeninas con asma. Se estimaron los costos directos anuales: visitas, pruebas de laboratorio, tratamiento farmacológico y de las crisis o exacerbaciones, para determinar la carga anual de la enfermedad desde una perspectiva institucional, y según la clasificación de la Iniciativa Global para el Asma. RESULTADOS: El costo promedio anual fue de $43,813,92, que aumentó en relación con la necesidad de aumento de dosis de corticoides inhalados y beta-agonistas de acción prolongada. El costo promedio de la consulta médica fue de $2004.57, $982.82 por gestión de crisis y $2645.95 por pruebas de laboratorio. El tratamiento farmacológico representó la principal carga económica, con un costo promedio anual de $38,180.58. CONCLUSIONES: Los resultados resaltan una carga económica del asma estimada en un costo anual por paciente de $43,813.92 MXN (DE=93,348.85), en el contexto del tercer nivel de atención en el sistema de salud público mexicano. La gravedad del asma, los tratamientos y los biológicos fueron los principales factores que aumentaron los costos directos de la atención.
OBJECTIVE: Determine the annual economic burden of the disease from an institutional perspective and based on GINA's recommended classification in a retrospective cohort of adults treated at Instituto Nacional de Enfermedades Respiratorias (INER) of Mexico City. METHODS: A retrospective, longitudinal observational study comprised by data from 247 female asthma patients, annual direct costs were estimated including: visits, laboratory tests, pharmacological treatment and management of crisis or exacerbations, to determine the annual burden of the disease from an institutional perspective and according to Global Initiative for Asthma classification. RESULTS: The average annual cost was $43,813.92, which increased in relation to the need of inhaled corticosteroids and long-acting beta agonists dosage increase. The average doctor's appointment cost was $2,004.57, $982.82 for crisis management and $2,645.95 for laboratory testing. Pharmacological treatment represented the main economic burden with an annual average cost of $38,180.58. CONCLUSIONS: The results highlight an economic burden of asthma estimated at an annual cost per patient of $43,813.92 MXN (SD=93,348.85) in the context of the third level of care in the Mexican public health system. The asthma severity and treatments such as biologics were the main factors that increased direct costs of care.
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Asma , Efeitos Psicossociais da Doença , Humanos , Asma/economia , Asma/tratamento farmacológico , Asma/terapia , México , Estudos Retrospectivos , Feminino , Adulto , Pessoa de Meia-Idade , Estudos Longitudinais , Academias e Institutos/economia , Adulto Jovem , Adolescente , IdosoRESUMO
ABSTRACT Magnesium (Mg) is essential for the metabolic reactions of the human body and is known for its biocompatibility, its mechanical and physical properties are similar to human bone, which is why it is considered to have high potential in biomedical applications such as temporary and resorbable implants. Through surface modifications, the high tendency to corrosion of Mg could be controlled, such as biodegradable membranes that prevent the passage of chloride ions present in the human organism. To prepare the membrane, solutions of chitosan modified with gelatin and/or glutaraldehyde are used and by means of the electrospray method applied to protect the Mg. To simulate body fluid conditions a Kokubo saline solution (BFK) was prepared. The study focuses on evaluating the corrosion rate of Mg with a coating made of a chitosan electrosprayed membrane, applying electrochemical measurements of electrochemical impedance spectroscopy and linear polarization resistance. The key additive to improve the behavior of the membranes was observed with the use of gelatin, where the membrane with the best results lowing corrosion rates is the Mg CH+GE+GL system, which it was observed with very good physical integrity in the images of morphological analyzes of the surface after 30 days of exposure.
RESUMEN El magnesio (Mg) es esencial para las reacciones metabólicas del cuerpo humano y es conocido por su biocompatibilidad, sus propiedades mecánicas y físicas son similares a las del hueso humano, por lo que se considera que tiene un alto potencial en aplicaciones biomédicas como implantes temporales y reabsorbibles. Mediante modificaciones superficiales se podría controlar la alta tendencia a la corrosión del Mg, como por ejemplo membranas biodegradables que impidan el paso de iones cloruro presentes en el organismo humano. Para preparar la membrana se utilizan soluciones de quitosano modificado con grenetina y/o glutaraldehído y mediante el método de electrorociado se aplican para proteger el Mg. Para simular las condiciones de los fluidos corporales se preparó una solución salina de Kokubo. El estudio se enfoca en evaluar la velocidad de corrosión del Mg con un recubrimiento hecho de una membrana electrorociada con quitosano, aplicando técnicas electroquímicas de espectroscopia de impedancia electroquímica y resistencia de polarización lineal. El aditivo clave para mejorar el comportamiento de las membranas se observó con el uso de gelatina, donde la membrana con mejores resultados bajando los índices de corrosión es el sistema Mg CH+GR+GL, el cual se observó con muy buena integridad física en las imágenes de análisis morfológicos de la superficie después de 30 días de exposición.
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Cholestasis, which is impaired bile flow from the liver into the intestine, can be caused by cholangitis and/or bile duct obstruction. Cholangitis can arise from bacterial infections and cholelithiasis, however, immune-mediated cholangitis in primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) is characterized by a strong immune response targeting the biliary epithelial cells (BECs). Persistent biliary inflammation further represents a risk for biliary neoplasia, cholangiocarcinoma (CCA) by driving chronic cellular stress in the BECs. Currently, immune-mediated cholangitis is considered a Th1-Th17-dominant disease, however, the presence of Th2-related mast cells (MCs) in tissue samples from PBC, PSC and CCA patients has been described, showing that these MCs are active players in these diseases. Here, we reviewed and discussed experimental and clinical data supporting a pro-fibrotic role for MCs in immune-mediated cholangitis as well as their participation in supporting tumor growth acting as angiogenesis promoters. Thus, although MCs have classically been identified as downstream effectors of Th2 responses in allergies and parasitic infections, evidence suggests that these MCs are relevant players in biliary inflammation and neoplasia. The availability of strategies to prevent MCs' activation represents a therapeutic opportunity in biliary diseases.
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Neoplasias dos Ductos Biliares , Colangiocarcinoma , Colangite Esclerosante , Colangite , Colestase , Neoplasias dos Ductos Biliares/patologia , Ductos Biliares Intra-Hepáticos/patologia , Colangiocarcinoma/patologia , Colangite/complicações , Colangite Esclerosante/patologia , Colestase/complicações , Humanos , Inflamação/complicações , Mastócitos/patologiaRESUMO
[This corrects the article DOI: 10.1155/2019/2056085.].
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Colitis-associated colorectal cancer (CRC) development has been shown to be related to chronically enhanced inflammation. Macrophage migration inhibitory factor (MIF) is an inflammatory mediator that favors inflammatory cytokine production and has chemotactic properties for the recruitment of macrophages (Møs) and T cells. Here, we investigated the role of MIF in the inflammatory response and recruitment of immune cells in a murine model of chemical carcinogenesis to establish the impact of MIF on CRC genesis and malignancy. We used BALB/c MIF-knockout (MIF-/-) and wild-type (WT) mice to develop CRC by administering intraperitoneal (i.p.) azoxymethane and dextran sodium sulfate in drinking water. Greater tumor burdens were observed in MIF-/- mice than in WT mice. Tumors from MIF-/- mice were histologically identified to be more aggressive than tumors from WT mice. The localization of MIF suggests that it is also involved in cell differentiation. The relative gene expression of il-17, measured by real-time PCR, was higher in MIF-/- CRC mice, compared to the WT CRC and healthy MIF-/- mice. Importantly, compared to the WT intestinal epithelium, lower percentages of tumor-associated Møs were found in the MIF-/- intestinal epithelium. These results suggest that MIF plays a role in controlling the initial development of CRC by attracting Møs to the tumor, which is a condition that favors the initial antitumor responses.
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Colite/metabolismo , Colite/patologia , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Fatores Inibidores da Migração de Macrófagos/metabolismo , Macrófagos/metabolismo , Linfócitos T/metabolismo , Animais , Progressão da Doença , Feminino , Citometria de Fluxo , Imuno-Histoquímica , Fatores Inibidores da Migração de Macrófagos/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos KnockoutRESUMO
OBJECTIVE: Some evidences indicate that exposure to molds or their products can be relevant for the loss of asthma control. Thus, we measured the mold burden present inside houses of subjects with asthma, and evaluated its relationship with asthma control. METHODS: Markers of asthma control in adult patients residing in Mexico City were evaluated through questionnaires and spirometry. Dust was collected from the patients' houses and its fungal content was determined by mold specific quantitative PCR (MSQPCR) for 36 fungal species. RESULTS: Forty-two patients with asthma (12 males, 30 females) with a mean age of 45 years (18-76 years) were included in the study. The level of asthma control measured through the Asthma Control Test ranged from 9 to 25 (mean 20.9). The FEV1/FVC ratio fluctuated from 38 to 106 %predicted (mean, 87.4 %predicted). Associations between mold burden and asthma control differed between males and females. Thus, concentrations of some molds, particularly Aspergillus fumigatus, Aureobasidium pullulans, Stachybotrys chartarum, Alternaria alternata, Cladosporium cladosporioides 2, Cladosporium herbarum, and Epicoccum nigrum, were negatively associated with parameters of asthma control in male subjects, but not in female patients. CONCLUSION: Our results showed that potential indoor exposure to some molds is associated with less asthma control in male subjects.
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Poluição do Ar em Ambientes Fechados/análise , Asma/microbiologia , Poeira/imunologia , Fungos/metabolismo , Adulto , Alternaria/metabolismo , Aspergillus fumigatus/metabolismo , Asma/fisiopatologia , Cladosporium/metabolismo , Feminino , Volume Expiratório Forçado , Fungos/crescimento & desenvolvimento , Habitação , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Espirometria/métodos , Stachybotrys/metabolismo , Avaliação de Sintomas/métodos , Avaliação de Sintomas/estatística & dados numéricos , Capacidade VitalRESUMO
The study of regulatory small RNAs, such as siRNAs and microRNAs in plants, has necessitated methods tailored to their unique features. Their analysis demands the use of sensitive and quantitative methods for their detection. The use of Northern blot hybridization offers an attractive alternative to address qualitative as well as quantitative features. We highlight the advantages and shortcomings of this method and offer a detailed description of the techniques that best work in our hands, considering their use for the study of several small RNAs in multiple samples. We enumerate relevant details as well as cautionary comments in cases where we have detected potential difficulties.
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MicroRNAs/genética , Plantas/genética , RNA de Plantas/genética , Northern Blotting/métodos , Regulação da Expressão Gênica de Plantas/genética , Hibridização de Ácido Nucleico/genética , RNA Interferente Pequeno/genéticaRESUMO
Plant microRNAs are commonly encoded in transcripts containing a single microRNA precursor. Processing by DICER-LIKE 1 and associated factors results in the production of a small RNA, followed by its incorporation into an AGO-containing protein complex to guide silencing of an mRNA possessing a complementary target sequence. Certain microRNA loci contain more than one precursor stem-loop structure, thus encoding more than one microRNA in the same transcript. Here, we describe a unique case where the evolutionary conserved miR398a is encoded in the same transcript as the legume-specific miR2119. The dicistronic arrangement found in common bean was also observed in other legumes. In Phaseolus vulgaris, mature miR398 and miR2119 are repressed in response to water deficit, and we demonstrate that both are functional as they target the mRNAs for CSD1 and ADH1, respectively. Our results indicate that the repression of miR398 and miR2119 leads to coordinated up-regulation of CSD1 and ADH1 mRNAs in response to water deficit in common bean and possibly in other legumes. Furthermore, we show that miRNA directed CSD1 and ADH1 mRNAs up-regulation also occurs when common bean plants are exposed to flooding, suggesting that plant redox status and fermentation metabolism must be closely coordinated under different adverse conditions.
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Álcool Desidrogenase/metabolismo , MicroRNAs/metabolismo , Phaseolus/metabolismo , Proteínas de Plantas/metabolismo , Precursores de RNA/metabolismo , Superóxido Dismutase/metabolismo , Álcool Desidrogenase/genética , Desidratação , Regulação da Expressão Gênica de Plantas/genética , MicroRNAs/genética , Phaseolus/fisiologia , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase , Precursores de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Superóxido Dismutase/genéticaRESUMO
BACKGROUND AND OBJECTIVE: Lung biopsies from patients with hypersensitivity pneumonitis (HP) have demonstrated small airway (SA) involvement, but there is no information concerning SA function in HP, and it is unknown whether pharmacological treatment could modify its function. SA function in patients with chronic HP using ultrasonic pneumography (UPG) and impulse oscillometry (IOS) was explored. We also compared initial results with those obtained after 4 weeks of standardized treatment with azathioprine and prednisone. METHODS: The study group consisted of adults with recent diagnoses of HP. All patients completed UPG, IOS, spirometry, body plethysmography, single-breath carbon monoxide diffusing capacity (DLCO ) and the 6-min walk test (6MWT). The fraction of exhaled nitric oxide (FENO ) was obtained to assess eosinophilic airway inflammation. Measurements were taken at diagnosis and after 4 weeks of treatment. RESULTS: A total of 20 consecutive patients (16 women) with chronic HP participated in the study. Median age was 50 years (interquartile range (IQR): 42-54). At diagnosis, the UPG phase 3 slope was abnormally high, consistent with maldistribution of ventilation. For IOS, all patients had low reactance at 5 Hz (X5) and elevated reactance area (AX) reflecting low compliance, and only eight (40%) patients had elevated R5 (resistance at 5 Hz (total)) and R5-20 (resistance at 5 Hz-resistance at 20 Hz (peripheral)) attributed to SA resistance. In contrast, FENO parameters were within normal limits. After treatment, forced vital capacity (FVC), the 6-min walk distance and the distribution of ventilation showed significant improvement, although DLCO did not. CONCLUSION: Patients with chronic HP have SA abnormalities that are partially revealed by the UPG and IOS tests. Lung volumes, but not gas exchange, improved after treatment with azathioprine and prednisone.
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Alveolite Alérgica Extrínseca , Azatioprina/farmacocinética , Pulmão , Prednisolona/farmacocinética , Resistência das Vias Respiratórias/fisiologia , Alveolite Alérgica Extrínseca/diagnóstico , Alveolite Alérgica Extrínseca/tratamento farmacológico , Alveolite Alérgica Extrínseca/fisiopatologia , Anti-Inflamatórios/farmacocinética , Disponibilidade Biológica , Testes Respiratórios/métodos , Feminino , Humanos , Pulmão/diagnóstico por imagem , Pulmão/fisiopatologia , Masculino , Pessoa de Meia-Idade , Oscilometria/métodos , Pletismografia/métodos , Testes de Função Respiratória/métodos , Espirometria/métodos , Volume de Ventilação Pulmonar/efeitos dos fármacos , Teste de Caminhada/métodosRESUMO
Fraction of exhaled nitric oxide (FeNO) is a marker of eosinophilic airway inflammation. Altitude above sea level can affect measurements of this index, but there is only limited information regarding the diurnal variation (ante meridiem vs. post meridiem) and reproducibility of FeNO on consecutive days at moderate altitudes. To evaluate the diurnal variability of FeNO and assess its reproducibility over five consecutive days in healthy individuals living at 2240 m, and to compare the FeNO readings taken with two different analyzers. Healthy non-smoking adults were measured using NIOX MINO(®) or NOA 280i(®) devices. One group (n = 10) had readings taken morning and afternoon for five consecutive days with the NIOX MINO(®) equipment; while the second group (n = 17) was measured on only one morning but by both the electrochemical analyzer (NIOX MINO(®)) and the chemiluminescence method (NOA 280i(®)). The study group consisted of 27 subjects aged 28.7 ± 6 years. Morning and afternoon FeNO measurements were 15.2 ± 7.5 ppb and 15.2 ± 7.9 ppb (p = 0.9), respectively. The coefficient of variation (CV) of these measurements (a.m. vs. p.m.) was 10.7 %, and the coefficient of repeatability (CR), 4.2 ppb. The concordance correlation coefficient (CCC) between the two measures (morning vs. afternoon) was 0.91. The CV and CR of the five morning readings were 15.4 % and 4.3 ppb, respectively; while those of the five afternoon measures were 13.6 % and 3.5 ppb, respectively. The CCC between the NIOX MINO(®) equipment and the NOA-280i(®) device was 0.8, with 95 % limits of agreement of -8.35 to 0.29 ppb. In adults living at 2240 m above sea level, FeNO measurements show minimal diurnal variation, and readings are reproducible (<15 %) over a period of at least five consecutive days; however, the FeNO measurements obtained with the NIOX MINO(®) and NOA 280i(®) devices are not interchangeable due to the wide limits of agreement recorded.
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Altitude , Óxido Nítrico/metabolismo , Adulto , Asma/metabolismo , Ritmo Circadiano , Estudos Transversais , Eosinofilia/metabolismo , Expiração , Feminino , Voluntários Saudáveis , Humanos , Masculino , Reprodutibilidade dos Testes , Adulto JovemRESUMO
Curcumin is a polyphenol and cisplatin is an antineoplastic agent that induces nephrotoxicity associated with oxidative stress, apoptosis, fibrosis and decrease in renal tight junction (TJ) proteins. The potential effect of curcumin against alterations in TJ structure and function has not been evaluated in cisplatin-induced nephrotoxicity. The present study explored whether curcumin is able to prevent the cisplatin-induced fibrosis and decreased expression of the TJ and adherens junction (AJ) proteins occludin, claudin-2 and E-cadherin in cisplatin-induced nephrotoxicity. Curcumin (200 mg kg(-1)) was administered in three doses, and rats were sacrificed 72 h after cisplatin administration. Curcumin was able to scavenge, in a concentration-dependent way, superoxide anion, hydroxyl radical, peroxyl radical, singlet oxygen, peroxynitrite anion, hypochlorous acid and hydrogen peroxide. Cisplatin-induced renal damage was associated with alterations in plasma creatinine, expression of neutrophil gelatinase-associated lipocalin and of kidney injury molecule-1, histological damage, increase in apoptosis, fibrosis (evaluated by transforming growth factor ß1, collagen I and IV and α-smooth muscle actin expressions), increase in oxidative/nitrosative stress (evaluated by Hsp70/72 expression, protein tyrosine nitration, superoxide anion production in isolated glomeruli and proximal tubules, and protein levels of NADPH oxidase subunits p47(phox) and gp91(phox), protein kinase C ß2, and Nrf2) as well as by decreased expression of occludin, claudin-2, ß-catenin and E-cadherin. Curcumin treatment prevented all the above-described alterations. The protective effect of curcumin against cisplatin-induced fibrosis and decreased proteins of the TJ and AJ was associated with the prevention of glomerular and proximal tubular superoxide anion production induced by NADPH oxidase activity.
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Junções Aderentes/efeitos dos fármacos , Antioxidantes/farmacologia , Cisplatino/toxicidade , Curcumina/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Junções Íntimas/efeitos dos fármacos , Animais , Antioxidantes/química , Biomarcadores , Curcumina/química , Fibrose/tratamento farmacológico , Sequestradores de Radicais Livres , Nefropatias/induzido quimicamente , Nefropatias/tratamento farmacológico , Masculino , NADPH Oxidases/química , NADPH Oxidases/genética , NADPH Oxidases/metabolismo , Proteína Quinase C beta/genética , Proteína Quinase C beta/metabolismo , Subunidades Proteicas , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , SuperóxidosRESUMO
BACKGROUND: MiRNAs and phasiRNAs are negative regulators of gene expression. These small RNAs have been extensively studied in plant model species but only 10 mature microRNAs are present in miRBase version 21, the most used miRNA database, and no phasiRNAs have been identified for the model legume Phaseolus vulgaris. Thanks to the recent availability of the first version of the common bean genome, degradome data and small RNA libraries, we are able to present here a catalog of the microRNAs and phasiRNAs for this organism and, particularly, we suggest new protagonists in the symbiotic nodulation events. RESULTS: We identified a set of 185 mature miRNAs, including 121 previously unpublished sequences, encoded by 307 precursors and distributed in 98 families. Degradome data allowed us to identify a total of 181 targets for these miRNAs. We reveal two regulatory networks involving conserved miRNAs: those known to play crucial roles in the establishment of nodules, and novel miRNAs present only in common bean, suggesting a specific role for these sequences. In addition, we identified 125 loci that potentially produce phased small RNAs, with 47 of them having all the characteristics of being triggered by a total of 31 miRNAs, including 14 new miRNAs identified in this study. CONCLUSIONS: We provide here a set of new small RNAs that contribute to the broader knowledge of the sRNAome of Phaseolus vulgaris. Thanks to the identification of the miRNA targets from degradome analysis and the construction of regulatory networks between the mature microRNAs, we present here the probable functional regulation associated with the sRNAome and, particularly, in N2-fixing symbiotic nodules.
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Phaseolus/genética , Proteínas de Plantas/genética , RNA de Plantas/análise , Análise de Sequência de RNA/métodos , Sequência Conservada , Bases de Dados Genéticas , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , MicroRNAs/análise , MicroRNAs/metabolismo , Phaseolus/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , RNA de Plantas/metabolismo , RNA Interferente Pequeno/análise , RNA Interferente Pequeno/metabolismoRESUMO
Interstitial lung diseases are a heterogeneous group of disorders that affect, to a greater or lesser degree, the alveolus, peripheral airway, and septal interstitium. Functional assessment in patients suspected of having an interstitial lung disease has implications for diagnosis and makes it possible to objectively analyze both response to treatment and prognosis. Recently the clinical value of lung-diffusing capacity and the six-minute walking test has been confirmed, and these are now important additions to the traditional assessment of lung function that is based on spirometry. Here we review the state-of-the-art methods for the assessment of patients with interstitial lung disease.
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Doenças Pulmonares Intersticiais/diagnóstico , Testes de Função Respiratória/métodos , Teste de Esforço/métodos , Humanos , Doenças Pulmonares Intersticiais/fisiopatologia , Doenças Pulmonares Intersticiais/terapia , Capacidade de Difusão Pulmonar/métodos , Espirometria/métodosRESUMO
The chemotherapeutic drug cisplatin has some side effects including nephrotoxicity that has been associated with reactive oxygen species production, particularly superoxide anion. The major source of superoxide anion is nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase. However, the specific segment of the nephron in which superoxide anion is produced has not been identified. Rats were sacrificed 72 h after cisplatin injection (7.5 mg/kg), and kidneys were obtained to isolate glomeruli and proximal and distal tubules. Cisplatin induced superoxide anion production in glomeruli and proximal tubules but not in distal tubules. This enhanced superoxide anion production was prevented by diphenylene iodonium, an inhibitor of NADPH oxidase. Consistently, this effect was associated with the increased expression of gp91(phox) and p47(phox), subunits of NADPH oxidase. The enhanced superoxide anion production in glomeruli and proximal tubules, associated with the increased expression of gp91(phox) and p47(phox), is involved in the oxidative stress in cisplatin-induced nephrotoxicity.
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Antineoplásicos/toxicidade , Cisplatino/toxicidade , Glomérulos Renais/efeitos dos fármacos , Túbulos Renais Proximais/efeitos dos fármacos , Glicoproteínas de Membrana/metabolismo , NADPH Oxidases/metabolismo , Superóxidos/metabolismo , Animais , Glomérulos Renais/metabolismo , Túbulos Renais Proximais/metabolismo , Masculino , NADPH Oxidase 2 , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
UNLABELLED: Cisplatin (CP) is an antineoplastic agent that induces nephrotoxicity and oxidative stress. It is unknown whether renal tight junction (TJ) proteins expression and localization are modified in CP-induced nephrotoxicity. OBJECTIVE: To study if the expression of the TJ proteins occludin, claudin-2, claudin-5 and zonula occludens-1 (ZO-1) is modified in rats with CP-induced nephrotoxicity. MATERIALS AND METHODS: Male Wistar rats (n = 5/group) were injected with saline solution (V group), and the other group (CP group) was injected with a single dose of saline solution and CP (7.5 mg/kg i.p.). Rats were sacrificed 72 h after CP injection and blood, and 24-h urine samples were collected. Several plasma and urinary injury biomarkers as well as renal histopathology lesions, oxidative and nitrosative stress markers were evaluated, and protein levels of ocludin, claudin-2, claudin-5, ZO-1 were measured by Western blot. Statistically significant changes noted with different p < 0.05 versus V. RESULTS: Nephrotoxicity was evident by histological alterations, glycosuria, decrease in creatinine clearance, increase in fractional excretion of sodium, serum creatinine and kidney injury molecule-1. These changes were associated with oxidative/nitrosative stress (increased renal abundance of 3-nitrotyrosine and protein kinase Cß2 and decreased renal expression of nuclear factor-erythroid-2-related factor 2) and decreased activity of antioxidant enzymes. Finally, it was found that CP-induced renal damage was associated with decreased renal expression of occludin and claudin-2. DISCUSSION AND CONCLUSION: CP altered the TJ proteins expression and localization in the proximal tubule that was associated with oxidative/nitrosative stress.
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Cisplatino/toxicidade , Rim/efeitos dos fármacos , Proteínas/metabolismo , Junções Íntimas/efeitos dos fármacos , Animais , Biomarcadores/sangue , Biomarcadores/urina , Western Blotting , Rim/metabolismo , Masculino , Ratos , Ratos Wistar , Junções Íntimas/metabolismoRESUMO
OBJECTIVES: Cisplatin (CP) is an antineoplastic agent that induces nephrotoxicity and oxidative stress. S-allylcysteine (SAC) is a garlic-derived antioxidant. This study aims to explore whether SAC protects against CP-induced nephrotoxicity in rats. METHODS: In the first stage, the SAC protective dose was determined by measuring renal damage and the oxidative stress markers malondialdehyde, oxidized proteins and glutathione in rats injected with CP. In the second stage, the effect of a single dose of SAC on the expression of nuclear factor-erythroid 2-related factor-2 (Nrf2), protein kinase C beta 2 (PKCß2) and nicotinamide adenine dinucleotide phosphate oxidase subunits (p47(phox) and gp91(phox) ) was studied. In addition, the effect of SAC on oxidative stress markers and on the activity of catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) in isolated proximal and distal tubules were evaluated. KEY FINDINGS: SAC (25 mg/kg) prevented the CP-induced renal damage and attenuated CP-induced decrease in Nrf2 levels and increase in PKCß2, p47(phox) and gp91(phox) expression in renal cortex and oxidative stress and decrease in the activity of CAT, GPx and GR in proximal and distal tubules. CONCLUSIONS: These data suggest that SAC provides renoprotection by attenuating CP-induced oxidative stress and decrease in the activity of CAT, GPx and GR.
Assuntos
Antioxidantes/uso terapêutico , Cisplatino/efeitos adversos , Cisteína/análogos & derivados , Alho/química , Nefropatias/prevenção & controle , Rim/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Antineoplásicos/efeitos adversos , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Catalase/metabolismo , Cisplatino/uso terapêutico , Cisteína/farmacologia , Cisteína/uso terapêutico , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Rim/metabolismo , Nefropatias/metabolismo , Masculino , Malondialdeído/metabolismo , Glicoproteínas de Membrana/metabolismo , NADPH Oxidase 2 , NADPH Oxidases/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Proteína Quinase C beta/metabolismo , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismoRESUMO
BACKGROUND: The PIKO-6® is an electronic device that measures forced expiratory volume at seconds 1 (FEV1) and 6 (FEV6) of a forced vital capacity (FVC) maneuver. This device could aid in diagnosing obstructive respiratory diseases. OBJECTIVES: To determine the concordance of FEV1, FEV6, and the FEV1/FEV6 quotient achieved with PIKO-6® versus spirometric values from asthmatic patients, and compare results with measures from healthy children. METHODS: A cross-sectional study with asthmatic and healthy 6-to-14-year-old children, all of whom performed a forced spirometry as well as a PIKO-6® test. RESULTS: The study included 82 subjects (58 asthmatics, 24 healthy children). Except for the functional parameters, the basal characteristics of the two groups were similar. The concordance correlation coefficient (CCC) for FEV1 was 0.938 (P < 0.001), with 95% limits of agreement of -0.591 to 0.512 L, and an average of differences of -0.040 L. For FEV6, CCC was 0.927 (P < 0.001), and the 95% limits of agreement were -0.751 to 0.598 L with an average of differences of -0.077 L. The concordance analysis and the FEV1 and FEV6 associations were better in children with controlled asthma and healthy subjects, as well as in the post-bronchodilator results. CONCLUSIONS: The concordance between PIKO-6® and spirometry was lower in patients with partially controlled or uncontrolled asthma compared to controlled or healthy children. The broad limits of agreement show that the FEV1, FEV6, and FEV1/FEV6 obtained with the PIKO-6® are not interchangeable with spirometry results. Longitudinal evaluations of asthma patients are necessary to assess the utility of PIKO-6®.
Assuntos
Asma/fisiopatologia , Volume Expiratório Forçado/fisiologia , Espirometria/instrumentação , Estudos de Casos e Controles , Criança , Estudos Transversais , Feminino , Humanos , Masculino , Reprodutibilidade dos Testes , Espirometria/métodos , Capacidade Vital/fisiologiaRESUMO
High-throughput sequencing techniques have been applied to the discovery of several different regulatory small RNAs, including siRNAs and microRNAs in different plant species. Their subsequent characterization demands the use of sensitive and quantitative methods for their detection. Here we describe the use of northern blot and quantitative PCR for these purposes. We highlight the advantages and shortcomings of each method and offer a detailed description of those techniques that best work in our hands, in particular having in mind their use for the study of several small RNAs in a given sample. We enumerate relevant alternatives as well as cautionary comments in cases where we have detected potential difficulties.
Assuntos
Fabaceae/genética , MicroRNAs/genética , Pequeno RNA não Traduzido/genética , Northern Blotting/métodos , RNA de Plantas , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Crop production of the important legume, the common bean (Phaseolus vulgaris), is often limited by low phosphorus (P) in the soil. The genotypes, BAT477 and DOR364, of the common bean have contrasting responses to P starvation. Plants from the BAT477 P deficiency tolerant genotype showed higher phosphate content and root biomass as compared to the DOR364 plants under P starvation. The PvPHR1 transcription factor-signaling pathway plays an essential role in the response to P starvation. PvPHO2, a negative regulator of this pathway, encodes an ubiquitin E2 conjugase that promotes degradation of P-responsive proteins and is the target gene of PvmiR399. PvPHO2 is downregulated in BAT477 plants under P deficiency, while such a response is not observed in P-starved DOR364 plants. Five putative PvmiR399 binding sites were identified in the 5' UTR region in both genotypes. While four sites showed an identical DNA sequence, the fifth (binding site of PvPHO2 one) showed three base changes and higher complementarity scores in DOR364 as compared to BAT477. Modified 5'RACE experiments indicated that PvmiR399 binding and/or processing was affected in DOR364 P-starved plants. We propose that a less efficient cleavage of the PvPHO2 mRNA directed by PvmiR399 would result in a higher PvPHO2-mediated degradation of P-responsive proteins in the DOR364 genotype with decreased P deficiency tolerance.