bZIP transcription factors PcYap1 and PcRsmA link oxidative stress response to secondary metabolism and development in Penicillium chrysogenum.

BACKGROUND: Reactive oxygen species (ROS) trigger different morphogenic processes in filamentous fungi and have been shown to play a role in the regulation of the biosynthesis of some secondary metabolites. Some bZIP transcription factors, such as Yap1, AtfA and AtfB, mediate resistance to oxidative stress and have a role in secondary metabolism regulation. In this work we aimed to get insight into the molecular basis of this regulation in the industrially important fungus Penicillium chrysogenum through the characterization of the role played by two effectors that mediate the oxidative stress response in development and secondary metabolism. RESULTS: In P. chrysogenum, penicillin biosynthesis and conidiation are stimulated by the addition of H2O2 to the culture medium, and this effect is mediated by the bZIP transcription factors PcYap1 and PcRsmA. Silencing of expression of both proteins by RNAi resulted in similar phenotypes, characterized by increased levels of ROS in the cell, reduced conidiation, higher sensitivity of conidia to H2O2 and a decrease in penicillin production. Both PcYap1 and PcRsmA are able to sense H2O2-generated ROS in vitro and change its conformation in response to this stimulus. PcYap1 and PcRsmA positively regulate the expression of brlA, the first gene of the conidiation central regulatory pathway. PcYap1 binds in vitro to a previously identified regulatory sequence in the promoter of the penicillin gene pcbAB: TTAGTAA, and to a TTACTAA sequence in the promoter of the brlA gene, whereas PcRsmA binds to the sequences TGAGACA and TTACGTAA (CRE motif) in the promoters of the pcbAB and penDE genes, respectively. CONCLUSIONS: bZIP transcription factors PcYap1 and PcRsmA respond to the presence of H2O2-generated ROS and regulate oxidative stress response in the cell. Both proteins mediate ROS regulation of penicillin biosynthesis and conidiation by binding to specific regulatory elements in the promoters of key genes. PcYap1 is identified as the previously proposed transcription factor PTA1 (Penicillin Transcriptional Activator 1), which binds to the regulatory sequence TTAGTAA in the pcbAB gene promoter. This is the first report of a Yap1 protein directly regulating transcription of a secondary metabolism gene. A model describing the regulatory network mediated by PcYap1 and PcRsmA is proposed.

Syndrome of professional wear and tear or «burnout¼ in Andalusian ophthalmology. / Síndrome de desgaste profesional o «burnout¼ en la oftalmología andaluza.

INTRODUCTION: Burnout syndrome refers to the physical and psychological fatigue suffered by workers as a result of their professional activity. It has three main characteristics: emotional exhaustion, depersonalization and lack of personal fulfillment. PURPOSE: To determine the prevalence of burnout syndrome among Andalusian ophthalmologists, both consultants and residents, analyzing its relationship with several sociodemographic variables. MATERIAL AND METHODS: Descriptive cross-sectional study. All the sociodemographic variables were included in a Google® form together with the standardized questionnaire Maslach Burnout Inventory-Human Services Survey (MBI-HSS). This form was sent to the Andalusian ophthalmological community by different electronic means (Andalusian Society of Ophthalmology, instant messaging mobile apps, etc.). RESULTS: One hundred fourty two ophthalmologists answered the form, with a similar distribution by sex and a mean age of 43.77 years, with 67.1% of them meeting criteria of burnout syndrome. Its prevalence was higher among young ophthalmologists, with fewer years of clinical practice, divorced/separated, those with fewer children, belonging to centers in the province of Huelva, workers in rural/regional areas, those with exclusive public activity, consultants with temporary contracts and four-year residents. Regarding subspecialties, its prevalence was higher among those who focused on cornea-ocular surface-refractive surgery. CONCLUSIONS: This study quantify for the first time in Andalusia the prevalence of burnout syndrome among ophthalmologists. The fact that two thirds of the analyzed sample meet criteria proves the need to implement improvements in the clinical and professional conditions of our group.

Molecular signatures of atherosclerotic plaques: An up-dated panel of protein related markers.

Atherosclerosis remains the leading cause of ischemic syndromes such as myocardial infarction or brain stroke, mainly promoted by plaque rupture and subsequent arterial blockade. Identification of vulnerable or high-risk plaques constitutes a major challenge, being necessary to identify patients at risk of occlusive events in order to provide them with appropriate therapies. Clinical imaging tools have allowed the identification of certain structural indicators of prone-rupture plaques, including a necrotic lipidic core, intimal and adventitial inflammation, extracellular matrix dysregulation, and smooth muscle cell depletion and micro-calcification. Additionally, alternative approaches focused on identifying molecular biomarkers of atherosclerosis have also been applied. Among them, proteomics has provided numerous protein markers currently investigated in clinical practice. In this regard, it is quite uncertain that a single molecule can describe plaque rupture, due to the complexity of the process itself. Therefore, it should be more accurate to consider a set of markers to define plaques at risk. Herein, we propose a selection of 76 proteins, from classical inflammatory to recently related markers, all of them identified in at least two proteomic studies analyzing unstable atherosclerotic plaques. Such panel could be used as a prognostic signature of plaque instability.

Toxicity of cadmium in musculoskeletal diseases.

Epidemiological studies have reported that exposure to toxic metals like cadmium (Cd) may promote the development of musculoskeletal diseases, such as osteoporosis, rheumatoid arthritis (RA), and osteoarthritis (OA), among others. The objective of this review is to summarize the molecular mechanisms of inflammation and oxidative stress activated by Cd at the bone level, particularly in osteoporosis, RA, and OA. Cadmium can increase bone resorption, affect the activity of osteoclasts and calcium (Ca) absorption, and impair kidney function, which favors the development of osteoporosis. In the case of RA, Cd interferes with the activity of antioxidant proteins, like superoxide dismutase (SOD) and catalase (CAT). It also promotes an inflammatory state, inducing the process of citrullination, which affects the proteins of immune response. On the other hand, accumulation of Cd in the tissues and blood of smokers has been related to the development of some musculoskeletal diseases. Therefore, knowing the negative impact of Cd toxicity at the articular level can help understand the damage mechanisms it produces, leading to the development of such diseases.

6-Methoxyquinoline complexes as lung carcinoma agents: induction of oxidative damage on A549 monolayer and multicellular spheroid model.

The aim of this work was to study the antitumor effects and the mechanisms of toxic action of a series of 6-methoxyquinoline (6MQ) complexes in vitro. The Cu(II) and Zn(II) complexes (Cu6MQ and Zn6MQ) are formulated as M(6MQ)2Cl2; the Co(II) and Ag(I) compounds (Co6MQ and Ag6MQ) are ionic with formulae [Ag(6MQ)2]+NO3- and H(6MQ)+[Co(6MQ)Cl3]- (where H(6MQ)+ is the protonated ligand). We found that the copper complex, outperformed the Co(II), Zn(II) and Ag(I) complexes with a lower IC50 (57.9 µM) in A549 cells exposed for 24 h. Cu6MQ decreased cell proliferation and induced oxidative stress detected with H2DCFDA at 40 µM, which reduces GSH/GSSG ratio. This redox imbalance induced oxidative DNA damage revealed by the Micronucleus test and the Comet assay, which turned into a cell cycle arrest at G2/M phase and induced apoptosis. In multicellular spheroids, the IC50 values tripled the monolayer model (187.3 µM for 24 h). At this concentration, the proportion of live/dead cells diminished, and the spheroids could not proliferate or invade. Although Zn6MQ also decreased GSH/GSSG ratio from 200 µM and the cytotoxicity is related to oxidative stress, the induction of the hydrogen peroxide levels only doubled the control value. Zn6MQ induced S phase arrest, which relates with the increased micronucleus frequency and with the induction of necrosis. Finally, our results reveal a synergistic activity with a 1:1 ratio of both complexes in the monolayer and multicellular spheroids.

Giant cell tumour of frontal bone in a patient with bilateral retinoblastoma. / Tumor óseo de células gigantes en techo orbitario de paciente con retinoblastoma bilateral.

A case is presented of a 5 year-old patient with bilateral hereditary retinoblastoma treated with radiotherapy in the right eye and enucleation of the left eye. After three years without evidence of progression, the patient presented with a right orbital mass that compromised the frontal bone. After surgical excision, the histology analysis was consistent with a diagnosis of giant cell tumour. Primary orbital bone neoplasms, such as giant cell tumours, are extremely rare. Both radiological and histopathology studies are essential to establish the differential diagnosis of orbital mass lesions.

[Attention deficit hyperactivity disorder in preschool age children. Its epidemiological prevalence in Navarra and La Rioja, Spain]. / Trastorno por deficit de atencion/hiperactividad en niños en edad preescolar. Prevalencia epidemiologica en Navarra y La Rioja, España.

INTRODUCTION: There are few studies about preschool attention deficit hyperactivity disorder (ADHD) prevalence. AIM: To study the prevalence of ADHD in preschoolers using an specific scale (ADHD-RS-IV-P-Es) developed for this age range. SUBJECTS AND METHODS: We evaluated the prevalence of possible ADHD in a representative sample of preschoolers in Navarra and La Rioja, Spain. RESULTS: We find a range of prevalence between 2.5-4.1% depending on the criteria that was used (more or less strict). CONCLUSIONS: There are specific tools that can be use in preschool ADHD study. The prevalence of preschool ADHD in Spain is similar than in other countries. The frequency of symptoms in this age range is similar to the found in school age children.

TITLE: Trastorno por deficit de atencion/hiperactividad en niños en edad preescolar. Prevalencia epidemiologica en Navarra y La Rioja, España.Introduccion. La prevalencia del trastorno por deficit de atencion/hiperactividad (TDAH) esta en constante estudio, y hay pocas publicaciones sobre la prevalencia en niños preescolares. Objetivo. Estudiar la prevalencia de sintomas de TDAH en niños preescolares usando una escala especifica (ADHD Rating Scale-IV, version preescolar, validada para España) desarrollada para esta franja de edad. Sujetos y metodos. Se evalua la prevalencia de posible TDAH en una muestra representativa de preescolares en Navarra y La Rioja usando diferentes puntos de corte. Resultados. Se encuentra un rango de prevalencia de sintomas de TDAH del 2,5-4,1% segun el criterio usado (mas o menos estricto). Conclusiones. Existen herramientas especificas que pueden usarse para evaluar el TDAH en preescolares. La prevalencia de posible TDAH en preescolares en España es similar a la encontrada en otros paises. La frecuencia de sintomas en esta franja de edad no es mas alta que en otras edades, por lo que no es cierto que los sintomas de TDAH sean muy prevalentes en la poblacion general de niños preescolares.

Psychomotor development and learning difficulties in preschool children with probable attention deficit hyperactivity disorder: An epidemiological study in Navarre and La Rioja. / Desarrollo psicomotor y dificultades del aprendizaje en preescolares con probable trastorno por déficit de atención e hiperactividad. Estudio epidemiológico en Navarra y La Rioja.

INTRODUCTION: ADHD symptoms begin to appear at preschool age. ADHD may have a significant negative impact on academic performance. In Spain, there are no standardized tools for detecting ADHD at preschool age, nor is there data about the incidence of this disorder. OBJECTIVE: To evaluate developmental factors and learning difficulties associated with probable ADHD and to assess the impact of ADHD in school performance. METHODS: We conducted a population-based study with a stratified multistage proportional cluster sample design. RESULTS: We found significant differences between probable ADHD and parents' perception of difficulties in expressive language, comprehension, and fine motor skills, as well as in emotions, concentration, behaviour, and relationships. Around 34% of preschool children with probable ADHD showed global learning difficulties, mainly in patients with the inattentive type. According to the multivariate analysis, learning difficulties were significantly associated with both delayed psychomotor development during the first 3 years of life (OR: 5.57) as assessed by parents, and probable ADHD (OR: 2.34) CONCLUSIONS: There is a connection between probable ADHD in preschool children and parents' perception of difficulties in several dimensions of development and learning. Early detection of ADHD at preschool ages is necessary to start prompt and effective clinical and educational interventions.

Programa integral para el cáncer en Cuba: Pautas para la gestión de implementación / Comprehensive program for cancer in Cuba: Guidelines for implementation management

Esta nueva edición sirve de orientación en la planificación y aplicación de intervenciones dirigidas a integrar el control del cáncer a las acciones del sistema nacional de salud, basa su actualización en el programa del médico y enfermera de la familia, y se estructura en paquetes tecnológicos como innovación para incorporar entre otros, los productos de la biotecnología. En ese sentido, organiza la gestión direccionada por el conocimiento, la socialización de las nuevas formas de organización establecidas para el control de la enfermedad.

Organic-Polymeric Radial Flow Biorreactor for Liver Models / Biorreactor de flujo radial orgánico-polimérico para modelos de hígado

Abstract An artificial liver support system is based on the functional hepatocytes being cultured inside a bioreactor; this technique has being used as an effective therapy for treating chronic liver diseases in recent times. This work evaluates different parameters such as cell viability and metabolic function of the hepatocytes when cultured on a hybrid scaffold. The scaffold was built using a polypyrrole plasma coated polymer layer seeded with endothelial matrix for efficient three-dimensional hepatocyte growth within a radial flow bioreactor. The flow rate inside the bioreactor was 7 ml / min. The parts for the bioreactor where either built using food-grade steel and/or glass or the scaffolds comprise a Poly (L-lactic acid)-coated polypyrrole iodine layer or not for HepG2 culture. The results show that the Poly (L-lactic acid)-coated scaffolds increased cell proliferation by 30%, protein production by 16% and albumin secretion by 40% compared with the non-coated scaffold. All experiments were performed thrice and data was analysed by ANOVA and Tukey statistic models with a p<0.05. The obtained results demonstrated that radial flow bioreactors in conjunction with hybrid scaffolds improve hepatocytes' physiological and functional properties and could be used as an alternative therapy for patients with liver diseases.

Resumen Un sistema de soporte hepático artificial se basa en utilizar hepatocitos funcionales cultivados en un biorreactor; esta técnica ha demostrado que se puede utilizar como una terapia eficaz para el tratamiento de enfermedades crónicas del hígado en los últimos tiempos. Este trabajo evalúa diferentes parámetros tales como la viabilidad celular y la función metabólica de los hepatocitos cuando se cultivan en un andamio híbrido. El andamio fue construido usando una capa de polímero recubierto de polipirrol plasma, se sembró con un cultivo tridimensional de células endoteliales y de hepatocitos dentro de un biorreactor de flujo radial. La velocidad de flujo en el interior del biorreactor fue de 7 ml / min. Las piezas para el biorreactor fueron construidas con acero de calidad alimentaria y / o vidrio. Los andamios control fueron de ácido L-poliláctico y a estos se les agrego un revestimiento de polipirrol-yodo para el cultivo de HepG2. Los resultados muestran que el ácido L-poliláctico recubierto, aumento la proliferación celular en un 30%, la producción de proteínas en un 16% y la secreción de albúmina por 40% en comparación con el andamio no recubierto. Todos los experimentos se llevaron a cabo tres veces y los datos se analizaron mediante modelos estadísticos ANOVA y Tukey con una p <0.05. Los resultados obtenidos demostraron que los biorreactores de flujo radial conjuntamente con andamios híbridos mejoran las propiedades fisiológicas y funcionales hepatocitos y podrían utilizarse como una terapia alternativa para los pacientes con enfermedades hepáticas crónicas.

Anti-inflammatory and antitumoural effects of Uncaria guianensis bark.

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria guianensis (Aublet) Gmell (Rubiaceae) is a medicinal plant from the jungles of South and Central America, used to treat cancer, arthritis, diabetes, and inflammation. Evaluate the anti-inflammatory and anti-tumor effects of Uncaria guianensis preparations. MATERIALS AND METHODS: Bio-guided fractionation of a hydroethanolic extract of Uncaria guianensis was performed, evaluating the fractions and subfractions for their effect on inflammatory mediators, tumour necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and prostaglandin E2 (PGE2) by ELISA and nitric oxide (NO) by the Griess reaction in cultured supernatant from RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). The expression of cyclooxygenase 2 (COX-2), inducible nitric oxide synthase (iNOS) and inhibitor of κB (IκB) were investigated in RAW 264.7 macrophages by flow cytometry. The activity of NF-κB in HeLa cells transfected with a luciferase reporter system was determined. The effect of Uncaria guianensis on the inflammatory response in vivo was assessed in BALB/c mice stimulated with LPS, on rat paw oedema induced by carrageenan, and on tumour growth and lung metastasis in BALB/c mice inoculated with 4T1 mammary tumour cells. Immune cell infiltrates and inflammatory mediators were evaluated in the tumour by immunohistochemistry. RESULTS: Sub-fraction Ug AIV inhibited, to varying degrees, NO, TNF-α, IL-6 and PGE2 production by macrophages in vitro (30 µg/ml) and in the serum of LPS-challenged mice (5 mg/kg). Macrophage expression of Cox-2 was inhibited (35%), IκB degradation was completely inhibited and NF-κB activation was inhibited (70%) by Ug AIV at 30 µg/ml. Ug AIV decreased paw oedema by 86% (5 mg/kg) and serum NO and TNF-α by 45% and 65% respectively. Ug AIV reduced 4T1 mammary tumour growth by 91% on day 33 post-inoculation as well as the levels of serum NO, IL-6 and TNF-α in the same animals. Ug AIV decreased the number of tumour-infiltrating T lymphocytes, macrophages and neutrophils as well as the number of cells positive for COX-2, iNOS, IL-6, TNF-α and p65. CONCLUSIONS: As Ug AIV was not cytotoxic for tumour cells or macrophages, its anti-tumour effect may be due to a reduction in pro-tumoural inflammatory processes in the tumour microenvironment, possibly mediated through NF-κB.

Socioeconomic deprivation does not affect prescribing of secondary prevention in patients with peripheral arterial disease.

AIM: Aim of the study was to assess the effect of socioeconomic deprivation on prescribing of cardiovascular secondary prevention medications in patients referred with peripheral arterial disease (PAD). METHODS: A retrospective review of vascular clinic referrals was performed. All patients referred from primary care with suspected PAD over a two month period were included. The deprivation score, prescription of cardiovascular secondary prevention medications, smoking status and the presence of cardiovascular co-morbidities (coronary artery or cerebrovascular disease--CAD/CVD) were assessed. Comparison was made between socioeconomic groups using the Carstairs Deprivation (DepCat) Score and between patients with and without a history of currently existing cardiovascular co-morbidities. RESULTS: The study included 391 patients. Almost two thirds of patients (253) were from the most deprived socioeconomic groups and were significantly younger at presentation (median age DepCat 7: 63 yrs, DepCat 1-2: 74.5 yrs, P<0.0001). The majority of patients with a prior history of CAD/CVD were prescribed secondary preventative medications at the time of referral with suspected PAD whereas those with no prior history of CAD/CVD, (212 patients, 54%) were significantly less likely to be prescribed antiplatelets (47% vs. 83%), statins (45% vs. 86%) or ACEi/ARBs (29% vs. 68%) (all P<0.05). Secondary prevention prescribing did not differ between socioeconomic groups. CONCLUSION: Secondary prevention prescribing is inadequate in patients with suspected PAD regardless of socioeconomic group and is significantly lower in those without previously diagnosed CAD/CVD. There remains a lack of appreciation of the high cardiovascular risk associated with PAD.

Valoración y Tratamiento de lesiones relaciones con la humedad y úlceras por presión: estudio piloto / Evaluation and Treatment of lesions related to moisture and pressure ulcers: pilot study

Las discrepancias entre autores respecto a la correcta diferenciación entre úlceras por presión (upp) y lesiones por humedad, así como las implicaciones que de ello se deriva, han favorecido la aparición de recientes líneas de trabajo entre grupos de expertos en el ámbito de los cuidados de la piel y las heridas. Tres van a ser los pilares básicos estructurales de este artículo: -la relevancia en términos de elección de estrategias preventivas y actitudes terapéuticas, la confirmada dificultad clínica en la diferenciación entre ambas entidades en la práctica asistencial diaria y el desconocimiento de muchos profesionales de enfermería sobre estas líneas de trabajo.Hemos elegido abordarlo bajo la perspectiva del caso clínico de una mujer de 96 años que presenta lesión por humedad entendiendo que se brinda una oportunidad para actualizar conocimientos bajo el deseo de contribuir a que las enfermeras ejecuten las medidas preventivas más adecuadas o proporcionen los cuidados enfermeros oportunos al deterioro de la integridad cutánea y/o tisular (AU)

There is a discrepancy between authors regarding the differentiation between pressure ulcer and lesions due to humidity, as well as their implications. This has raised new research lines concerned with the care of the skin and wounds. This review has three basic sections: the importance of selecting appropriate preventive and therapeutic strategies, the confirmed clinical difficulty in differentiation between the two entities in the daily medical practice, and the unawareness of many nurses about these new research lines. We present a clinical case of a 96-year-old woman with skin lesions due to humidity. The described management of this case gives the opportunity to update the knowledge about the most appropriate preventive measures and nursing care to avoid he deterioration of the skin integrity in case of moisture (AU)

Hepatocytes display a compensatory survival response against cadmium toxicity by a mechanism mediated by EGFR and Src.

Although the liver is a cadmium-target organ, hepatocyte response involved in its toxicity is not yet elucidated. A link between this heavy metal treatment and Stat3 signaling pathways was examined in primary mouse hepatocytes. We provided evidence of a novel link among NADPH oxidase and Stat3 signaling, mediated by Src, EGFR, and Erk1/2. Cadmium activates NADPH oxidase. ROS produced by this oxidase activates Src, enable that in turn, transactivates EGFR that activates Stat3 in tyrosine, allowing its dimerization. Also, ROS from NADPH oxidase favors ERK1/2 activation that phosphorylates Stat3 in serine, resulting in a compensatory or adaptive survival response such as production of metallothionein-II in short Cd exposure times. However, after 12h CdCl2 treatment, cell viability diminished in 50%, accompanied by a drastic decrease of metallothionein-II production, and an increase in p53 activation and the pro-apoptotic protein Bax.