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IL-9 is a pleiotropic cytokine associated with various processes, including antitumor immunity, induction of allergic pathologies, and the immune response against helminth infections, where it plays an important role in the expulsion of the parasite. In a murine model of Nippostrongylus brasiliensis infection, IL-9 is produced mainly by CD4+ T lymphocytes and innate lymphoid cells found in the lung, small intestine, and draining lymph nodes. Given the technical difficulties involved in the intracellular staining of IL-9, as well as the complexity of isolating hematopoietic cells from the small intestine upon infection, there is a pressing need for a comprehensive but straightforward protocol to analyze the expression of IL-9 in different lymphoid and non-lymphoid tissues in this model. The protocol described here outlines the kinetics of IL-9 produced by CD4+ T cells and innate lymphoid cells in the lung and small intestine, the main organs targeted by N. brasiliensis, as well as in the mediastinal and mesenteric lymph nodes, throughout the infection. In addition, it details the number of larvae needed for infection, depending on the cell type and organ of interest. This protocol aims to assist in the standardization of assays to save time and resources by offering the opportunity to focus on the specific cells, organs, and disease stages of interest in the N. brasiliensis infection model.

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Ras-GTPases are nucleotide hydrolases involved in key cellular processes. In fungi, Ras-GTPases regulate conidiation, development, virulence, and interactions with other fungi or plants. Trichoderma spp. are filamentous saprophytic fungi, widely distributed along all latitudes, characterized by their rapid growth and metabolic diversity. Many species of this genus interact with other fungi, animals or plants. Furthermore, these fungi are used as biocontrol agents due to their ability to antagonize phytopathogenic fungi and oomycetes, through competence, antibiosis, and parasitism. However, the genetic and molecular regulation of these processes is scarcely described in these fungi. In this work, we investigated the role of the gene tbrg-1 product (GenBank accession number XP_013956100; JGI ID: Tv_70852) of T. virens during its interaction with other fungi and plants. Sequence analyses predicted that TBRG-1 bears the characteristic domains of Ras-GTPases; however, its size (1011 aa) is 3- to 4-times bigger compared with classical GTPases. Interestingly, phylogenetic analyses grouped the TBRG-1 protein with hypothetical proteins of similar sizes, sharing conserved regions; whereas other known Ras-GTPases were perfectly grouped with their respective families. These facts led us to classify TBRG-1 into a new family of Ras-GTPases, the Big Ras-GTPases (BRG). Therefore, the gene was named tbrg-1 (TrichodermaBigRas-GTPase-1). Quantification of conidia and scanning electron microscopy showed that the mutants-lacking tbrg-1 produced less conidia, as well as a delayed conidiophore development compared to the wild-type (wt). Moreover, a deregulation of conidiation-related genes (con-10, con-13, and stuA) was observed in tbrg-1-lacking strains, which indicates that TBRG-1 is necessary for proper conidiophore and conidia development. Furthermore, the lack of tbrg-1 affected positively the antagonistic capability of T. virens against the phytopathogens Rhizoctonia solani, Sclerotium rolfsii, and Fusarium oxysporum, which was consistent with the expression patterns of mycoparasitism-related genes, sp1 and cht1, that code for a protease and for a chitinase, respectively. Furthermore, the antibiosis effect of mycelium-free culture filtrates of Δtbrg-1 against R. solani was considerably enhanced. The expression of secondary metabolism-related genes, particularly gliP, showed an upregulation in Δtbrg-1, which paralleled an increase in gliotoxin production as compared to the wt. These results indicate that TBRG-1 plays a negative role in secondary metabolism and antagonism. Unexpectedly, the biocontrol activity of Δtbrg-1 was ineffective to protect the tomato seeds and seedlings against R. solani. On the contrary, Δtbrg-1 behaved like a plant pathogen, indicating that TBRG-1 is probably implicated in the recognition process for establishing a beneficial relationship with plants.

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While first discovered in immunoreceptor signaling, the Syk protein kinase behaves as a tumor and metastasis suppressor in epithelial cells. Its reduced expression in breast and other carcinomas is correlated with decreased survival and increased metastasis risk, but its action mechanism remains largely unknown. Using phosphoproteomics we found that Syk phosphorylated E-cadherin and α-, ß-, and p120-catenins on multiple tyrosine residues that concentrate at intercellular junctions. Increased Syk expression and activation enhanced E-cadherin/catenin phosphorylation, promoting their association and complex stability. In human breast cancer cells, Syk stimulated intercellular aggregation, E-cadherin recruitment and retention at adherens junctions, and promoted epithelial integrity, whereas it inhibited cell migration and invasion. Opposite effects were obtained with Syk knockdown or non-phosphorylatable mutant E-cadherin expression. Mechanistically, Syk stimulated the interaction of the E-cadherin/catenin complex with zonula occludens proteins and the actin cytoskeleton. Conditional Syk knockout in the lactating mouse mammary gland perturbed alveologenesis and disrupted E-cadherin localization at adherens junctions, corroborating the observations in cells. Hence, Syk is involved in the maintenance of the epithelial integrity of the mammary gland via the phosphorylation and stabilization of the E-cadherin/catenin adherens junction complex, thereby inhibiting cell migration and malignant tumor invasion.

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Trichoderma spp. are a rich source of secondary metabolites and volatile organic compounds (VOCs), which may induce plant defenses and modulate plant growth. In filamentous fungi, chromatin modifications regulate secondary metabolism. In this study we investigated how the absence of histone deacetylase HDA-2 in the Trichoderma atroviride strain Δhda-2 impacts its effect on a host, Arabidopsis (Arabidopsis thaliana). The production of VOCs and their impact on plant growth and development were assessed as well. The Δhda-2 strain was impaired in its ability to colonize Arabidopsis roots, thus affecting the promotion of plant growth and modulation of plant defenses against foliar pathogens Botrytis cinerea and Pseudomonas syringae, which normally result from interaction with T. atroviride Furthermore, Δhda-2 VOCs were incapable of triggering plant defenses to counterattack foliar pathogens. The Δhda-2 overproduced the VOC 6-pentyl-2H-pyran-2-one (6-PP), which resulted in enhanced root branching and differentially regulated phytohormone-related genes. Analysis of ten VOCs (including 6-PP) revealed that three of them positively regulated plant growth, whereas six had the opposite effect. Assessment of secondary metabolites, detoxification, and communication with plant-related genes showed a dual role for HDA-2 in T. atroviride gene expression regulation during its interaction with plants. Chromatin immunoprecipitation of acetylated histone H3 on the promoters of plant-responsive genes in Δhda-2 showed, in the presence of Arabidopsis, low levels of epl-1 and abc-2 compared with that in the wild type; whereas ctf-1 presented high constitutive levels, supporting a dual role of HDA-2 in gene regulation. This work highlights the importance of HDA-2 as a global regulator in Trichoderma to modulate multiple responses in Arabidopsis.

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Objetivos. Conocer los motivos por los que los pacientes ingresados en neumología continúan fumando, y describir las características de fumadores, exfumadores y no fumadores. Métodos. Estudio observacional de corte transversal de 6 meses de duración. Se entregó una encuesta anónima y voluntaria a todos los pacientes ingresados en hospitalización. Resultados. Fueron incluidos un total de 284 pacientes, de los cuales 57 (20,1%) eran fumadores, 125 (44%) exfumadores y 102 (35,9%) no habían fumado nunca. Entre los fumadores hombres (37), el motivo más frecuente por el que fumaban fue ‘Placer-relajación’ (78,38%), mientras que entre las fumadoras mujeres (20) fue ‘Reducción de estados negativos’ (90%). Los exfumadores (125) dejaron de fumar en su mayoría por ‘Progresión de su enfermedad’ (50,4%). Los exfumadores presentaban una mayor frecuencia de hipertensión arterial, cardiopatía, enfermedad pulmonar obstructiva crónica (EPOC) y neoplasias que los fumadores, y mayor frecuencia de cardiopatías, EPOC y neoplasias que en aquellos que nunca habían fumado. El riesgo de padecer EPOC fue mayor en fumadores al compararlo con los que nunca habían fumado. Las mujeres comenzaron a fumar más tarde con una tasa de tabaquismo acumulado menor que los hombres. Una mayor edad se mostró como un predictor positivo de abandono. Conclusiones. Un 20,1% de los pacientes fumaba en el momento del ingreso. Continúan fumando por placer-relajación, adicción y reducción de estados negativos. Se ha encontrado en exfumadores una mayor frecuencia de diversas patologías, siendo la EPOC más frecuente en fumadores. Mayor edad se mostró como predictor positivo de abandono (AU)

Objectives. Determine the reasons why respiratory inpatients continue to smoke and describe the characteristics of smokers, former smokers and people who have never smoked. Methods. It was a 6-month observational, transversal study. A questionnaire was given to every patient admitted in the respiratory department. Results. 284 patients were included in the study, of which 57 (20.1%) were smokers, 125 (44%) former smokers and 102 (35.9%) had never smoked. Amongst the smoker men (37), the most frequent reason to continue smoking was ‘Pleasurable relaxation’ (78.38%), while in smoker women (20) it was ‘Tension reduction’ (90%). Former smokers (125) quitted the habit due to ‘Progression of their disease’ (50.4%). Former smokers had a greater frequency of developing hypertension, heart disease, chronic obstructive pulmonary disease (COPD) and cancer than those who had never smoked. The risk of having COPD was greater in smokers compared to those who had never smoked. Women began smoking later with a lower accumulated smoking rate than men. An older age was shown to be a positive predictor of abandon. Conclusions. A 20.1% of patients were reported as smokers at the time of admission. In general they continue smoking due to relaxation and pleasure, addiction and tension reduction. Different types of pathologies were found more frequently in former smokers, although COPD was more frequent in smokers. An older age was shown to be a positive predictor of abandon (AU)

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Objetivo. Conocer la percepción del estado emocional de las mujeres con embarazo en vías de prolongación. Metodología. Se llevó a cabo una investigación con metodología cualitativa descriptiva, fundamentada en entrevistas individuales en profundidad a siete gestantes en vías de prolongación, con edades entre los 21 y 32 años y una duración del embarazo comprendida entre 40+5 y 41+4. Resultados. El estado emocional varía a lo largo del embarazo, una vez pasada la fecha probable de parto. En este sentido, el bienestar emocional se ve alterado por el miedo que es genera el dolor del parto, a las posibles complicaciones y al cuidado del recién nacido, por lo que presentan un estado de nerviosismo por el parto inminente. Las mujeres, además, necesitan de la presencia de un acompañante durante el parto y después del mismo. Conclusión. El miedo puede afectar negativamente la experiencia subjetiva del embarazo en la fase final del mismo, al proceso de trabajo del parto y a la transición hacia la maternidad en las gestantes con embarazo en vías de prolongación. Estas mujeres necesitan el apoyo y educación específica por parte de la enfermera matrona según sus necesidades individuales...

Objective. This study sought to know the perception of the emotional state of women with pregnancy susceptible to prolongation. Methodology. The research was conducted with qualitative descriptive methodology, founded on in-depth individual interviews of 7 gestants susceptible to prolongation, with ages between 21 and 32 years and duration of the pregnancy comprised between 40+5 and 41+4 weeks. Results. The emotional state varies throughout the pregnancy; once the probable date of delivery has passed, the emotional well-being is altered by fear of pain during delivery, possible complications, and caring for the newborn, presenting a state of nervousness due to the imminent delivery. In addition, the women described the need for the presence of a companion during and after the delivery. Conclusion. Fear can affect negatively the subjective experience of the pregnancy during its final phase, the delivery process, and the transition to maternity in gestants with pregnancy susceptible to prolongation. These women need support and specific education from the midwife according to their individual needs...

Objetivo. Conhecer a percepção do estado emocional das mulheres com gravidez em via de prolongamento. Metodologia. Levou-se a cabo uma investigação com metodologia qualitativa descritiva, fundamentada em entrevistas individuais em profundidade a 7 gestantes em via de prolongamento, com idades entre os 21 e 32 anos e uma duração da gravidez compreendida entre 40+5 e 41+4. Resultados. O estado emocional varia ao longo da gravidez, uma vez passada a data provável de parto, o bem-estar emocional se vê alterado pelo medo à dor do parto, às possíveis complicações e ao cuidado do recém nascido, apresentando um estado de nervosismo pelo parto iminente. As mulheres ademais descrever a necessidade da presença de um acompanhante durante o parto e depois do mesmo. Conclusão. O medo pode afetar negativamente à experiência subjetiva da gravidez na fase final do mesmo, ao processo de trabalho do parto e à transição para a maternidade nas gestantes com gravidez em via de prolongamento. Estas mulheres precisam o apoio e educação específica por parte da enfermeira matrona segundo suas necessidades individuais...

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@#<p style="text-align: justify;"><strong>OBJECTIVES:</strong> The study aimed to determine the frequency and clinical profile of nosocomial infections among pediatric patients with Acute Lymphoblastic Leukemia (ALL) admitted at the Philippine General Hospital from January 2010 to December 2011.<br /><strong>METHODOLOGY:</strong> This is a descriptive cross sectional study involving the review of medical records of pediatric patients with ALL under the charity service of the Section of Pediatric Hematology and Oncology of Philippine General Hospital (PGH). The frequency, clinical presentation, outcome, causes of death and isolated organisms from sterile sites were determined. Fischer's Exact test was used to assess correlation between variables with Gram-negative bacterial infection and mortality.<br /><strong>RESULTS:</strong> There were 80 documented nosocomial infections among 45 pediatric patients with ALL. The incidence was 9.22 per 100 admissions. The majority presented initially with fever (50%) and 37 (82%) had neutropenia. Blood stream infections were present in 33 (73%) out of 45 pediatric ALL patients. Gram-negative organisms (76%), particularly Pseudomonas putida (33%), Pseudomonas aeruginosa (12%) and Burkholderia pseudomallei (12%) were isolated in majority of patients. About 42.2% of the 45 patients had resolution of the infection and 26.7% resulted to death primarily caused by septic shock in 7 (58.3%) of the 12 patients identified. Presence of fever (p- value 0.011, RR 2.1094) was associated with presence of Gram-negative bacteria at 5% level of significance and with 2.109 times more risk to having a gram-negative infection. There was no significant correlation between mortality and symptoms of infection.<br /><strong>CONCLUSION:</strong> Nosocomial infections remain to be significant cause of morbidity and death among pediatric patients with ALL. Gram negative infections were the most common cause of sepsis in these patients. One fourth of the patients with nosocomial infections died.</p>

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The purpose of the present study was to explore the usefulness of the Mexican sequential organ failure assessment (MEXSOFA) score for assessing the risk of mortality for critically ill patients in the ICU. A total of 232 consecutive patients admitted to an ICU were included in the study. The MEXSOFA was calculated using the original SOFA scoring system with two modifications: the PaO2/FiO2 ratio was replaced with the SpO2/FiO2 ratio, and the evaluation of neurologic dysfunction was excluded. The ICU mortality rate was 20.2%. Patients with an initial MEXSOFA score of 9 points or less calculated during the first 24 h after admission to the ICU had a mortality rate of 14.8%, while those with an initial MEXSOFA score of 10 points or more had a mortality rate of 40%. The MEXSOFA score at 48 h was also associated with mortality: patients with a score of 9 points or less had a mortality rate of 14.1%, while those with a score of 10 points or more had a mortality rate of 50%. In a multivariate analysis, only the MEXSOFA score at 48 h was an independent predictor for in-ICU death with an OR = 1.35 (95%CI = 1.14-1.59, P < 0.001). The SOFA and MEXSOFA scores calculated 24 h after admission to the ICU demonstrated a good level of discrimination for predicting the in-ICU mortality risk in critically ill patients. The MEXSOFA score at 48 h was an independent predictor of death; with each 1-point increase, the odds of death increased by 35%.

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El uso de esteroide está reconocido en el tratamiento crítico del paciente quemado y es útil en el choque séptico que no responde a vasopresores. Este grupo de medicamentos ayuda a regular la respuesta hemodinámica mejorando el aporte de sangre a la piel, aunque sabemos que tienen un efecto nocivo en el proceso de cicatrización. Evaluamos el efecto histopatológico del empleo de esteroides en quemaduras usando un modelo animal. Empleamos 2 grupos de 10 ratas (Wistar) en las que colocamos un cilindro de metal en el dorso durante 15 segundos a 95oC, produciendo una quemadura. En ese momento, uno de los grupos recibió esteroide a dosis de estrés (hidrocortisona 5 mg/kg) y el otro no recibió ningún medicamento. Al quinto día resecamos la escara y cubrimos el defecto con cultivo de queratinocitos. Los animales fueron sacrificadas a los 14 días. Realizamos análisis histopatológico. Macroscópicamente evaluamos la (..) (AU)

The use of steroids is well recognized in critical care specially in septic shock. There are some reports of their utility in severe burns. It helps to regulate the hemodynamic response in order to improve the blood supply to the skin, although it is well known their negative effect in wound healing. Our objective is to know the hystopathologic effect of steroids in burn healing. We used 2 groups of 10 rats (Wistar). Both groups were exposed in their backs to a metallic cylinder at 95 oC for 15 seconds. At the moment of the burn, one group was given steroid (hydrocortisone at stress dose 5 mg/kg) and the other group didn't received any medication. The scar was removed at the 5th day and the burn injury was covered with queratinocyte culture. The rats were sacrificed at 14th day. We evaluated the presence of clinical signs of infection and the percentage of new epithelium. In the microscope we evaluated the following parameters: fibrosis, inflammatory process, presence of fibroblast and vascular proliferation. We compared both groups using Chi2 test (SPSS program version 10). A p =/<. 05 was considered as statistical significant. We found no difference between each group in fibrosis (p .47), inflammatory process (p .27), or fibroblast presence (p.16). But there was a difference in vascular proliferation (p .05) against the first group (steroid group). There were no signs of infection and all of them were epithelised at the 14th day. In conclusion, the use of steroids in burns in an animal model could have a final effect in wound healing. In humans it is important to say that they can be helpful in those cases with clear evidence of benefit, as for example failure to vassopresor response in septic shock. We are not sure about the final effect in wound healing in the steroid group as for example wound contracture in long term (AU)

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Objetivo: determinar la incidencia de estreñimiento en la Provinciade Albacete y su relacion con la dieta y los estilos de vida.Material y métodos: Encuesta transversal poblacional a 414pacientes encuestados mayores de 50 años en la provincia de Albacete.Se incluyeron en el estudio 445 personas mayores de 50 añosseleccionadas por un muestreo aleatorio sistemático, contestaron correctamente414. Mediciones principales: edad, peso y talla; estadocivil, nivel de estudios y ocupación habitual del paciente; padecimientode enfermedad; horas que duerme al día; ejercicio físico; consumode tabaco; consumo de alcohol; consumo de medicamentos (antiinflamatoriosy laxantes); hábitos intestinales; régimen o dieta; frecuenciade las comidas y lugar donde come habitualmente; frecuencia deconsumo de alimentos a la semana; consumo de agua diario, café, téy otras infusiones; toma de complejos vitamínicos y suplementos defibra; antecedentes familiares con cáncer.Resultados: de las personas que participaron en nuestro estudio,56,9% eran mujeres con una edad media de 67,07 años.La mayoría de los encuestados realizan tres comidas diarias, desayuno,comida y cena, mientras que el 50% realizan una comida amedia mañana o la merienda. Estas comidas las realizan habitualmenteen el propio domicilio. Predominó el consumo de leche(83,7%), pan (95,1%), verduras (68,8%), frutas (91,8%) y aceite deoliva virgen (96,6%) diariamente, con consumo de pescados cada 1-2 días así como las legumbres y consumo de carne cada 3-6 días.Un 44,4% de encuestados consumían entre 1-2 litros de agua aldía. Tan sólo un 3,9% de la muestra suplementaba su dieta con algúncomplejo vitamínico. Un 35% seguían algún tipo de dieta o régimen.Se observó que el 97,7% de los encuestados con más de 3deposiciones semanales tenían un consumo alto de aceite de olivavirgen. En nuestra encuesta realizan ejercicio físico habitualmente el65,7% de la muestra. De las personas del estudio, el 70,2% no fueronfumadores, mientras que el 10,2% eran fumadores diarios y un18,4% ex-fumadores. Respecto al alcohol, de los 414 encuestadosel porcentaje de bebedores fue de un 35,1%. El principal tipo demedicación que tomaban los encuestados fueron los AINE con unporcentaje de 14,5% de la muestra. Un 79,7% no tomaba ni AINEni laxantes. Sólo un 2,7% de los encuestados consumían laxanteshabitualmente.Conclusiones: Existe una mayoría con hábitos alimenticiosmás o menos sanos

Objective: to determine the incidence of constipation in Albaceteprovince and its relation with diet and lifestyle.Patients and methods: cross-sectional population survey.We studied 414 participants over 50 years of age in Albaceteprovince. 445 persons over 50 years of age were included in thestudy. All participants were selected by systematic random sampling;414 participants filled in the questionnaire correctly. Mainmeasures: age, weight and height, marital status, level of educationand occupation; presence of a disease, number of sleep hoursa day, physical exercise, smoking, alcohol intake, drug intake(anti-inflammatories and laxatives); bowel habit, diet, meal frequencyand place; food intake frequency per week, daily intake ofwater, coffee, tea and herbal beverages; vitamin and fiber supplements;presence of cancer in the family.Results: 56.9% of participants were women. Mean age67.07 years. In Albacete province, 4.4% of the population over50 years have a bowel habit consistent with constipation. Mostparticipants had three meals a day (breakfast, lunch, and supper),while 50% had another meal in the morning or afternoon.These meals took place, habitually, in the domicile. There was apreponderance in daily intake of the following foods: milk(83.7%), bread (95.1%), vegetables (68.8%), fruit (91.8%), andvirgin olive oil (96.6%). Fish was eaten every one to two days,and pulses and meat every three to six days. 44.4% of participantsdrank one to two liters of water a day. Only 3.9% of participantstook some supplement; 35% of participants were on adiet. It was observed that 97.7% of participants with more thanthree defecations a week had a high intake of virgin olive oil;65.7% of participants did some physical exercise customarily;70.2% of participants were non-smokers, 10.2% were smokers,and 18.4% were ex-smokers. With regard to alcohol, the percentageof drinkers was 35.1%. The main class of medicationstaken by participants was NSAIDs –14.5%; 79.7% took neitherNSAIDs nor laxatives. Only 2.7% of participants took laxativesregularly.Conclusions: Most participants had relatively healthy eatinghabits

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Objetivo. Valorar la relación de los diferentes factores epidemiológicos con la insuficiencia venosa crónica (IVC) en una zona básica de salud (ZBS) de atención primaria. Pacientes y métodos. 4.857 Pacientes censados en la ZBS de Valverde del Júcar, en la provincia de Cuenca; se selecciona una muestra de 477 pacientes en torno a tres grupos de riesgo: grupo de profesionales carpinteros, grupo de amas de casa y grupo de jubilados, y se captan a través de divulgación informativa, contacto con líderes de los grupos y visita en lugares de trabajo. El estudio se desarrolla durante 6 meses. Resultados. 477 personas se incluyeron en la primera visita, de las cuales 352 finalizan el estudio en ésta por no reunir criterios de IVC (clasificación CEAP de IVC). Se diagnostican 125 personas de IVC, y se confirman mediante prueba con Doppler. La proporción hombre/mujer fue de 3:2, la edad media fue de 41,20 ñ 15,28 años para hombres y 53,31 ñ 19,11 años para mujeres, donde tanto unos como otros desempeñan una actividad laboral mayoritariamente de pie. Los factores de riesgo para IVC más frecuentes fueron: profesión de riesgo (90,4 por ciento), sobrepeso (76,8 por ciento) y antecedentes familiares. Conclusiones. Se refleja una asociación no estadísticamente significativa entre el desarrollo de IVC y los factores epidemiológicos (AU)

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Streptococcus mutans is a pathogen related to the occurrence of human dental caries. The determination of total amounts of mutans streptococci, as well as the proportion related to other oral bacteria, is of interest when assessing the risk of developing caries. In this context, it is better to use a sensitive, specific and non-time consuming method such as the polymerase chain reaction (PCR), than to use culture and biochemical identification methods. In this work we identified potentially cariogenic strains of S. mutans and assessed the relationship with the dmft, DMFT or dmft/DMFT index. Using DNA isolated from dental plaque, a 192 bp sequence was identified and amplified from the spaP gene and a 722 bp sequence from the dexA gene. The results suggest that it is important to evaluate the presence of cariogenic S. mutans strains in plaque content rather than the accumulation of plaque itself However, other factors like diet, hygiene, genetic background, the flow rate of saliva and the presence of specific antibodies, also play a key role in the development of caries.

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A two-session experiment is proposed to train students with an easy, economic and educative procedure to detect the typical DNA ladder produced in many apoptotic events. The procedure is accurate enough to provide an easy way to compare degrees of damage in DNA caused by different treatments.

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